Updated on 2024/12/12

写真a

 
TANAKA Takaaki
 
Organization
Academic Assembly Institute of Science and Technology SEISAN DESIGN KOUGAKU KEIRETU Professor
Graduate School of Science and Technology Advanced Materials Science and Technology Professor
Faculty of Engineering Department of Engineering Professor
Title
Professor
External link

Degree

  • 博士(農学) ( 1996.3   京都大学 )

  • 工学修士 ( 1989.3   京都大学 )

Research Interests

  • porous materials

  • 分離材料

  • (bio)mechanical evaluation

  • bioengineering

  • membrane

  • Biofunction Bioprocess

Research Areas

  • Manufacturing Technology (Mechanical Engineering, Electrical and Electronic Engineering, Chemical Engineering) / Transport phenomena and unit operations

  • Nanotechnology/Materials / Structural materials and functional materials

  • Life Science / Applied microbiology

  • Manufacturing Technology (Mechanical Engineering, Electrical and Electronic Engineering, Chemical Engineering) / Biofunction and bioprocess engineering

  • Life Science / Applied biochemistry

Research History

  • Niigata University   Faculty of Engineering Department of Engineering   Professor

    2017.4

  • Niigata University   Graduate School of Science and Technology Advanced Materials Science and Technology   Professor

    2013.4

  • Niigata University   Graduate School of Science and Technology Advanced Materials Science and Technology   Professor

    2013.4

  • Niigata University   Faculty of Engineering Department of Material Science and Technology   Professor

    2013.4 - 2017.3

  • Niigata University   Faculty of Engineering Department of Material Science and Technology   Associate Professor

    2004.4 - 2013.3

  • Niigata University   Faculty of Engineering Department of Material Science and Technology   Associate Professor (as old post name)

    1999.1 - 2004.3

  • Niigata University   Faculty of Engineering Department of Material Science and Technology   Lecturer

    1996.5 - 1998.12

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Professional Memberships

 

Papers

  • Preparation of composite monoliths of quaternized chitosan and diatom earth for protein separation Reviewed

    Takaaki Tanaka, Yuna Tomita, Koki Honda, Marino Fujisawa, Akihito Ochiai

    Journal of Separation Science   46 ( 2 )   2200638 - 2200638   2023.1

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    Authorship:Lead author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Wiley  

    DOI: 10.1002/jssc.202200638

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    Other Link: https://onlinelibrary.wiley.com/doi/full-xml/10.1002/jssc.202200638

  • Preparation of polyhydroxyalkanoate microfiltration membranes via nonsolvent–induced phase separation methods Reviewed

    Kazuyoshi Tabata, Toshiki Shibuya, Kousuke Karakida, Akihito Ochiai, Masayuki Taniguchi, Takaaki Tanaka

    MEMBRANE   45 ( 6 )   315 - 323   2020.11

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  • Platelet adhesion on commercially pure titanium plates in vitro III: effects of calcium phosphate-blasting on titanium plate biocompatibility Reviewed

    Nakamura, Masayuki, Aizawa, Hachidai, Kawabata, Hideo, Sato, Atsushi, Watanabe, Taisuke, Isobe, Kazushige, Kitamura, Yutaka, Tanaka, Takaaki, Kawase, Tomoyuki

    International Journal of Implant Dentistry   6   74-1 - 74-14   2020.11

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  • Poly(L-lactic acid) depth filter membrane prepared by nonsolvent-induced phase separation with the aid of a nonionic surfactant Reviewed

    H. Minbu, H. Mizuno, Y. Shibuya, A. Ochiai, M. Taniguchi, T. Tanaka

    Journal of Chemical Engineering of Japan   52 ( 1 )   75 - 82   2019.1

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    DOI: 10.1252/jcej.18we084

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  • Mechanical and degradation properties of advanced platelet-rich fibrin (A-PRF), concentrated growth factors (CGF), and platelet-poor plasma-derived fibrin (PPTF) Reviewed

    Isobe Kazushige, Watanebe Taisuke, Kawabata Hideo, Kitamura Yutaka, Okudera Toshimitsu, Okudera Hajime, Uematsu Kohya, Okuda Kazuhiro, Nakata Koh, Tanaka Takaaki, Kawase Tomoyuki

    INTERNATIONAL JOURNAL OF IMPLANT DENTISTRY   3   2017.5

  • Biomechanical evaluation by AFM of cultured human cell-multilayered periosteal sheets Reviewed

    Makoto Horimizu, Tomoyuki Kawase, Takaaki Tanaka, Kazuhiro Okuda, Masaki Nagata, Douglas M. Burns, Hiromasa Yoshie

    Micron   48   1 - 10   2013.5

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    We previously demonstrated that thicker periosteal sheets with enhanced cell layering maintain their component cells at relatively immature stages of differentiation but express a high in vivo osteogenic potential. As it has been recently proposed that stiff scaffolds provide a mechanical cue to various cell types that promotes differentiation, we postulated that the maintenance of immature cells in our periosteal sheets is due to the mechanical stiffness of the multilayered-cell architecture. To demonstrate the biomechanical characteristics of our periosteal sheets, we have determined their stiffnesses with atomic force microscopy (AFM) and evaluated the expression of extracellular matrix (ECM) components specifically by both immunocytochemistry and a complementary DNA microarray technology. Compared to osteoblastic Saos2 cells, the cytoskeletal fibers were developed more in the periosteal cells, but the periosteal cells in monolayer culture developed before either the cells in the peripheral or central regions of the periosteal sheets developed. However, the nanoindentation by AFM distinguished the central region from the peripheral region. The peak stiffness values of cells were ordered as follows: tissue culture polystyrene (1.66. GPa). ⋙. dispersed (9.99. kPa). &gt
    . central region (5.20. kPa). &gt
    . peripheral regions (3.67. kPa). Similarly, the degree of development of α-smooth muscle actin (αSMA) filaments within cells was dispersed. &gt
    . central region. &gt
    . peripheral region. In conjunction with the abundantly deposited ECM in the periosteal sheets, these findings suggest that the order of cell stiffness may depend on the integration of the stiffness of individual ECM components and the extent of cytoskeletal fiber formation. Because recently published data have demonstrated that the optimal stiffness for osteogenic differentiation is 25-40. kPa, it is plausible that the periosteal cells residing in the less-stiff multilayer regions could be maintained at relatively immature stages under the control of the stem-cell medium in vitro but start differentiating when exposed to the proper stiffness upon release from the culture conditions at the implantation site. © 2013 Elsevier Ltd.

    DOI: 10.1016/j.micron.2013.02.001

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  • Effects of SARS‑CoV‑2 mRNA vaccines on platelet polyphosphate levels and inflammation: A pilot study

    Takashi Uematsu, Atsushi Sato, Hachidai Aizawa, Tetsuhiro Tsujino, Taisuke Watanabe, Kazushige Isobe, Hideo Kawabata, Yutaka Kitamura, Takaaki Tanaka, Tomoyuki Kawase

    Biomedical Reports   16 ( 3 )   2022.2

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    Publishing type:Research paper (scientific journal)   Publisher:Spandidos Publications  

    DOI: 10.3892/br.2022.1504

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  • Crystal structure of rice defensin OsAFP1 and molecular insight into lipid-binding Reviewed

    Akihito Ochiai, Kodai Ogawa, Minami Fukuda, Masami Suzuki, Kosuke Ito, Takaaki Tanaka, Yoshiyuki Sagehashi, Masayuki Taniguchi

    Journal of Bioscience and Bioengineering   130 ( 1 )   6 - 13   2020.7

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    Defensins are antibacterial peptides that function in the innate immune system. OsAFP1, a defensin identified from Oryza sativa (rice), exhibits antimicrobial activity against rice pathogens. Intriguingly, OsAFP1 was also shown to demonstrate potent antifungal activity against the human pathogenic fungus Candida albicans by inducing apoptosis in target cells, suggesting that OsAFP1 represents a potential new antibiotic candidate; however, further analyses, particularly at the structural level, are required to elucidate the mechanistic underpinnings of OsAFP1 antifungal activity. Here, we determined the three-dimensional structure of OsAFP1 using X-ray crystallography. OsAFP1 features the cysteine-stabilized αβ structure highly conserved in plant defensins and presents a dimeric structure that appears necessary for antifungal activity. Superimposition of the OsAFP1 structure with that of Nicotiana alata NaD1 complexed with phosphatidic acid indicated that the target molecule is likely trapped between the S2-S3 loops of each OsAFP1 dimer. In lipid-binding analyses performed using nitrocellulose membranes immobilized with various membrane lipid components, OsAFP1 was found to bind to phosphatidylinositols (PIPs) harboring phosphate groups, particularly PI(3)P. These results indicate that OsAFP1 exerts antifungal activity by binding to PI(3)P contained in the C. albicans cell membrane, thereby applying cellular stress and inducing apoptosis. Furthermore, the OsAFP1 structure and site-specific-mutation analyses revealed that Arg1, His2, Leu4, Arg9, and Phe10 play critical roles in OsAFP1 dimer formation. Thus, our study provides novel insights into the antifungal mechanism of OsAFP1.

    DOI: 10.1016/j.jbiosc.2020.02.011

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  • Identification of cationic peptides derived from low protein rice by-products and evaluation of their multifunctional activities. Reviewed

    Taniguchi M, Aida R, Saito K, Oya R, Ochiai A, Saitoh E, Tanaka T

    Journal of Bioscience and Bioengineering   129 ( 3 )   307 - 314   2020.3

  • Identification and characterization of multifunctional cationic peptides from enzymatic hydrolysates of soybean proteins. Reviewed

    Masayuki Taniguchi, Ryousuke Aida, Kazuki Saito, Toyotaka Kikura, Akihito Ochiai, Eiichi Saitoh, Takaaki Tanaka

    Journal of Bioscience and Bioengineering   129 ( 1 )   59 - 66   2020.1

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    In this study, we used the commercial soybean protein hydrolysate Hinute-DC6 as a novel starting material from which to purify and identify multifunctional cationic peptides. After fractionation, Hinute-DC6 was separated into 20 fractions with varying isoelectric points (pI) by ampholyte-free isoelectric focusing (autofocusing). Subsequently, we purified and identified the cationic peptides from fractions 19 and 20, which had pI values greater than 12, using reversed-phase high-performance liquid chromatography and matrix-assisted laser/desorption ionization-time-of-flight mass spectrometry. Of the 83 cationic peptides identified, 14 had high pI values and net charges greater than +2, and were chemically synthesized and assayed for various bioactivities, including hemolytic, antimicrobial, lipopolysaccharide (LPS)-neutralizing, and angiogenic activities. None of the 14 cationic peptides tested exhibited hemolytic activity toward mammalian red blood cells at concentrations up to 1000 μM. Five of the cationic peptides exhibited antimicrobial activities against at least one of four human-pathogenic microorganisms tested. In addition, in chromogenic LPS-neutralizing assays using Limulus amebocyte lysates, the 50% effective concentrations of these 14 peptides were between 0.069 and 5.2 μM. Tube-formation assays in human umbilical vein endothelial cells showed that each of the 14 cationic peptides exhibited significant angiogenic activities at 10 μM, with values similar to those of the positive control LL-37. Our results demonstrate that the 14 identified cationic peptides have multiple functions with negligible hemolytic activity. These data indicate that the cationic peptides isolated from Hinute-DC6 and fractions containing these cationic peptides have the potential to be used as multifunctional ingredients for healthcare applications.

    DOI: 10.1016/j.jbiosc.2019.06.016

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  • Wound healing activity and mechanism of action of antimicrobial and lipopolysaccharide-neutralizing peptides from enzymatic hydrolysates of rice bran proteins. Reviewed

    Masayuki Taniguchi, Kazuki Saito, Ryousuke Aida, Akihito Ochiai, Eiichi Saitoh, Takaaki Tanaka

    Journal of Bioscience and Bioengineering   128 ( 2 )   142 - 148   2019.8

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    In our previous study, we identified multifunctional cationic peptides from enzymatic hydrolysates of rice bran proteins (RBPs) that have antimicrobial and lipopolysaccharide-neutralizing activities. In this study, we investigated the potential of the peptides RBP-LRR, RBP-EKL, and RBP-SSF to promote proliferation, angiogenesis (tube formation), and migration in human umbilical vein endothelial cells (HUVECs). To determine mechanisms of wound healing actions, angiogenic and migration-promoting activities of these peptides were evaluated following pretreatments of HUVECs with specific inhibitors. In these experiments, the cationic peptides RBP-LRR, RBP-EKL, and RBP-SSF induced cell proliferation at low concentrations of 0.1 μM or 1 μM. Moreover, the three cationic peptides had angiogenic activities at concentrations more than 1 μM in tube formation assays, and their effects were similar to those of LL-37. Subsequent scratch migration assays exhibited that RBP-LRR, RBP-EKL, and RBP-SSF promote wound closure at optimum concentrations of 10, 10, and 0.1 μM, respectively. In further studies, we performed tube formation assays using HUVECs pretreated with SU5416, which inhibits vascular endothelial growth factor (VEGF) receptors, and suggested the possibility that the three cationic peptides induce angiogenesis by activating VEGF receptors. In corresponding scratch migration assays using HUVECs, pretreatment with the proliferation inhibitor mitomycin C did not alter the effects of RBP-LRR and RBP-EKL, and significant contribution to wound closure were mediated by cell migration regardless of proliferation rates. In contrast, RBP-SSF contributed to wound closure exclusively by promoting cell proliferation. The present data indicate that RBP-LRR, RBP-EKL, and RBP-SSF are candidates for use as wound healing agents.

    DOI: 10.1016/j.jbiosc.2019.02.002

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  • Evidence for contamination of silica microparticles in advanced platelet-rich fibrin matrices prepared using silica-coated plastic tubes Reviewed

    T. Tsujino, A. Takahashi, S. Yamaguchi, T. Watanabe, K. Isobe, Y. Kitamura, T. Tanaka, K. Nakata, T. Kawase

    Biomedicines   7 ( 2 )   45-1 - 45-11   2019.6

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    DOI: 10.3390/biomedicines7020045

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  • Protein adsorption characteristics of nanoparticle-assembled hollow microspheres of hydroxyapatite and their composites with PLLA microporous membranes Reviewed

    T. Tanaka, Y. Takai, A. Nagase, K. Teraguchi, H. Minbu, A. Ochiai, I. Kimura, M. Taniguchi

    Heliyon   5   e01490-1 - e01490-22   2019.4

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    DOI: 10.1016/j.heliyon.2019.e01490

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  • Identification and characterization of multifunctional cationic peptides from traditional Japanese fermented soybean Natto extracts. Reviewed

    Masayuki Taniguchi, Ryousuke Aida, Kazuki Saito, Akihito Ochiai, Satoshi Takesono, Eiichi Saitoh, Takaaki Tanaka

    Journal of Bioscience and bioengineering   127 ( 4 )   472 - 478   2019.4

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    In this study, we investigated the lipopolysaccharide (LPS)-neutralizing and angiogenic activities of cationic peptides derived from the traditional Japanese fermented product Natto, which is made by fermenting cooked soybeans using Bacillus subtilis. Initially, we prepared 20 fractions of Natto extracts with various isoelectric points (pI's) using ampholyte-free isoelectric focusing (autofocusing). Cationic peptides were then purified from fractions 19 and 20, whose pH values were greater than 12, using reversed-phase high-performance liquid chromatography, and were identified using matrix-assisted laser/desorption ionization-time-of-flight mass spectroscopy. Among the 13 identified cationic peptides, seven (KFNKYGR, FPFPRPPHQK, GQSSRPQDRHQK, QRFDQRSPQ, ERQFPFPRPPHQK, GEIPRPRPRPQHPE, and EQPRPIPFPRPQPR) had pI's greater than 9.5, positive net charges, and differing molecular weights. These peptides were then chemically synthesized and applied to chromogenic LPS-neutralizing assays using Limulus amebocyte lysates, and 50% effective (neutralizing) concentrations of 2.6-5.5 μM were demonstrated. In addition, tube formation assays in human umbilical vein endothelial cells revealed angiogenic activities for all but one (GEIPRPRPRPQHPE) of these seven cationic peptides, with increases in relative tube lengths of 23-31% in the presence of peptides at 10 μM. Subsequent experiments showed negligible hemolytic activity of these peptides at concentrations of up to 500 μM in mammalian red blood cells. Collectively, these data demonstrate that six cationic peptides from Natto extracts, with the exception of GEIPRPRPRPQHPE, have LPS-neutralizing and angiogenic activities but do not induce hemolysis.

    DOI: 10.1016/j.jbiosc.2018.09.016

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  • Cationic peptides from enzymatic hydrolysates of soybean proteins exhibit LPS-neutralizing and angiogenic activities. Reviewed

    Masayuki Taniguchi, Yusuke Noda, Ryousuke Aida, Kazuki Saito, Akihito Ochiai, Eiichi Saitoh, Takaaki Tanaka

    Journal of bioscience and bioengineering   127 ( 2 )   176 - 182   2019.2

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    In this study, we prepared fractions containing multifunctional cationic peptides by separating the commercial soybean protein hydrolysate Hinute-AM into 20 fractions. These fractions contained peptides with various isoelectric points (pI), as indicated by ampholyte-free isoelectric focusing (autofocusing). Thus, we purified and identified the cationic peptides from fractions 19 and 20, which had pH values greater than 10, using reversed-phase high-performance liquid chromatography and matrix-assisted laser/desorption ionization-time-of-flight mass spectroscopy. Among 19 identified cationic peptides, NKNAKPPSPR, PGKKNAIV, KSGPGMSPR, NVSKPPRVV, RKVGAGGRKPLG, and LPCVIGGVPKRV had high pI values and were included as chemically synthesized peptides in assays of various functions, including lipopolysaccharide (LPS)-neutralizing and angiogenic activities. Chromogenic LPS-neutralizing assays using Limulus amebocyte lysates showed that 50% effective concentrations of these six peptides were between 1.63 and 2.65 μM, and were higher than that (0.12 μM) of polymyxin B. Moreover, in tube-formation assays in human umbilical vein endothelial cells, all of the six cationic peptides except LPCVIGGVPKRV exhibited angiogenic activities similar to those of the positive control LL-37. In addition, the six identified cationic peptides had no hemolytic activity at concentrations up to 500 μM in mammalian red blood cells. Our results demonstrate that five of the identified cationic peptides, excluding LPCVIGGVPKRV, have multiple functions and little or no hemolytic activity. These data indicate that fractions containing cationic peptides from Hinute-AM have the potential to be used as dietary supplements and functional ingredients in food products.

    DOI: 10.1016/j.jbiosc.2018.07.013

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  • Spectrophotometric determination of platelet counts in platelet-rich plasma Reviewed

    Y. Kitamura, M. Suzuki, T. Tsukioka, K. Isobe, T. Tsujino, T. Watanabe, T. Watanabe, H. Okudera, K. Nakata, T. Tanaka, T. Kawase

    International Journal of Implant Dentistry   4   29-1 - 29-8   2018.10

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    DOI: 10.1186/s40729-018-0140-8

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  • Rice Defensin OsAFP1 is a New Drug Candidate against Human Pathogenic Fungi Reviewed

    Ochiai Akihito, Ogawa Kodai, Fukuda Minami, Ohori Masahiro, Kanaoka Takumi, Tanaka Takaaki, Taniguchi Masayuki, Sagehashi Yoshiyuki

    SCIENTIFIC REPORTS   8   2018.7

  • The antimicrobial and anti-endotoxic peptide AmyI-1-18 from rice α-amylase and its [N3L] analog promote angiogenesis and cell migration. Reviewed International journal

    Masayuki Taniguchi, Akihito Ochiai, Toshiki Namae, Kazuki Saito, Tetsuo Kato, Eiichi Saitoh, Takaaki Tanaka

    Peptides   104   78 - 84   2018.6

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    In our previous studies, we showed that AmyI-1-18 and its single amino acid-substituted analogs have antimicrobial, anti-inflammatory, and anti-endotoxic activities and cause little or no hemolysis or cytotoxicity. In this study, we investigated the potential of these peptides to promote proliferation, angiogenesis (tube formation), and migration in human umbilical vein endothelial cells (HUVECs). Among five single amino acid-substituted analogs, [N3L]AmyI-1-18 induced cell proliferation in a concentration-dependent manner with similar efficacy to AmyI-1-18. In tube formation assays, AmyI-1-18 and [N3L]AmyI-1-18 had angiogenic activities at 1 μM and their effects were similar to those of LL-37. Moreover, scratch migration assays showed that AmyI-1-18, [N3L]AmyI-1-18, and LL-37 promote cell migration with optimum concentrations of 10, 1, and 0.1 μM, respectively. Subsequently, we performed tube formation assays using HUVECs pretreated with SU5416, which is an inhibitor of vascular endothelial growth factor (VEGF) receptors, and revealed that AmyI-1-18 and [N3L]AmyI-1-18 induce angiogenesis by activating VEGF receptors. Similarly, after pretreating HUVECs with mitomycin C, which inhibits cell proliferation, [N3L]AmyI-1-18 significantly contributed to wound closure in scratch migration assays. Moreover, enhancements of hydrophobicity following substitution of AmyI-1-18 asparagine with leucine led to greater increases in cell migration. The present data indicate that both peptides, particularly [N3L]AmyI-1-18, are candidates for use as wound healing agents.

    DOI: 10.1016/j.peptides.2018.04.017

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  • Platelet counts in insoluble platelet-rich fibrin clots: A direct method for accurate determination Reviewed

    Yutaka Kitamura, Taisuke Watanabe, Masayuki Nakamura, Kazushige Isobe, Hideo Kawabata, Kohya Uematsu, Kazuhiro Okuda, Koh Nakata, Takaaki Tanaka, Tomoyuki Kawase

    Frontiers in Bioengineering and Biotechnology   6   2018.2

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    Platelet-rich fibrin (PRF) clots have been used in regenerative dentistry most often, with the assumption that growth factor levels are concentrated in proportion to the platelet concentration. Platelet counts in PRF are generally determined indirectly by platelet counting in other liquid fractions. This study shows a method for direct estimation of platelet counts in PRF. To validate this method by determination of the recovery rate, whole-blood samples were obtained with an anticoagulant from healthy donors, and platelet-rich plasma (PRP) fractions were clotted with CaCl2 by centrifugation and digested with tissue-plasminogen activator. Platelet counts were estimated before clotting and after digestion using an automatic hemocytometer. The method was then tested on PRF clots. The quality of platelets was examined by scanning electron microscopy and flow cytometry. In PRP-derived fibrin matrices, the recovery rate of platelets and white blood cells was 91.6 and 74.6%, respectively, after 24 h of digestion. In PRF clots associated with small and large red thrombi, platelet counts were 92.6 and 67.2% of the respective total platelet counts. These findings suggest that our direct method is sufficient for estimating the number of platelets trapped in an insoluble fibrin matrix and for determining that platelets are distributed in PRF clots and red thrombi roughly in proportion to their individual volumes. Therefore, we propose this direct digestion method for more accurate estimation of platelet counts in most types of platelet-enriched fibrin matrix.

    DOI: 10.3389/fbioe.2018.00004

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  • An updated proposal for terminology and classification of platelet-rich fibrin Reviewed

    Tomoyuki Kawase, Takaaki Tanaka

    REGENERATIVE THERAPY   7   80 - 81   2017.12

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    DOI: 10.1016/j.reth.2017.10.002

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  • Cationic peptides from peptic hydrolysates of rice endosperm protein exhibit antimicrobial, LPS-neutralizing, and angiogenic activities Reviewed

    Masayuki Taniguchi, Junya Kawabe, Ryu Toyoda, Toshiki Namae, Akihito Ochiai, Eiichi Saitoh, Takaaki Tanaka

    PEPTIDES   97   70 - 78   2017.11

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    In this study, we hydrolyzed rice endosperm protein (REP) with pepsin and generated 20 fractions containing multifunctional cationic peptides with varying isoelectric point (pI) values using ampholyte-free isoelectric focusing (autofocusing). Subsequently, we determined antimicrobial activities of each fraction against the pathogens Prophyromonas gingivalis, Propionibacterium acnes, Streptocossus mutans, and Candida albicans. Fractions 18, 19, and 20 had pI values greater than 12 and exhibited antimicrobial activity against P. gingivalis, P. acnes, and C. albicans, but not against S. mutans. In further experiments, we purified and identified cationic peptides from fractions 18, 19, and 20 using reversed-phase high-performance liquid chromatography and matrix-assisted laser/desorption ionization-time-of-flight mass spectroscopy. We also chemically synthesized five identified peptides (RSVSKSR, RRVIEPR, ERFQPMFRRPG, RVRQNIDNPNRADTYNPRAG, and VVRRVIEPRGLL) with pI values greater than 10.5 and evaluated antimicrobial, lipopolysaccharide (LPS)-neutralizing, and angiogenic activities. Among these synthetic peptides, only VVRRVIEPRGLL exhibited antimicrobial activity against P. gingivalis, with an IC50 value of 87 mu M. However, all five cationic peptides exhibited LPS-neutralizing and angiogenic activities with little or no hemolytic activity against mammalian red blood cells at functional concentrations. These present data show dual or multiple functions of the five identified cationic peptides with little or no hemolytic activity. Therefore, fractions containing cationic peptides from REP hydrolysates have the potential to be used as dietary supplements and functional ingredients in food products.

    DOI: 10.1016/j.peptides.2017.09.019

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  • Identification and characterization of multifunctional cationic peptides derived from peptic hydrolysates of rice bran protein Reviewed

    Masayuki Taniguchi, Mitsuhiro Kameda, Toshiki Namae, Akihito Ochiai, Eiichi Saitoh, Takaaki Tanaka

    JOURNAL OF FUNCTIONAL FOODS   34   287 - 296   2017.7

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    In this study, to prepare the fraction containing multifunctional cationic peptides, we first hydrolyzed rice bran protein (RBP) with pepsin. We separated the enzymatic hydrolysate of RBP into 20 fractions containing peptides with different isoelectric point (pI) values by ampholyte-free isoelectric focusing (autofocusing). Subsequently, we examined the antimicrobial activity of each fraction against four pathogens. In addition, we purified the cationic peptides from fractions exhibiting antimicrobial activity by reversed phase high-performance liquid chromatography and identified them by matrix-assisted laser/desorption ionization-time-of-flight mass spectroscopy. Of five cationic peptides identified, we chemically synthesized three peptides with high pI values and evaluated their multiple functions, including antimicrobial, lipopolysaccharide-neutralizing and angiogenic activities. Our results demonstrated that the three identified cationic peptides exhibited multiple functions with little or no haemolytic activity. Fractions containing cationic peptides obtained from RBP hydrolysate have the potential to be used as dietary supplements and functional ingredients in food products. (C) 2017 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.jff.2017.04.046

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  • Identification and characterization of multifunctional cationic and amphipathic peptides from soybean proteins Reviewed

    Masayuki Taniguchi, Kengo Saito, Takafumi Nomoto, Toshiki Namae, Akihito Ochiai, Eiichi Saitoh, Takaaki Tanaka

    BIOPOLYMERS   108 ( 4 )   2017.7

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    In this study, we identified and chemically synthesized three cationic and amphipathic peptides (Glycinin-17, BCAS-16, and BCBS-11) from soybean proteins. These peptides had high isoelectric points, high positive net charges, and included multiple hydrophobic amino acids. Subsequently, we identified multiple functions of these peptides, including antimicrobial, lipopolysaccharide-neutralizing, and angiogenic activities, and examined their cytotoxic activities against mammalian red blood cells. Glycinin-17, BCAS-16, and BCBS-11 exhibited antimicrobial activity against Porphyromonas gingivalis and Candida albicans whereas Glycinin-17 did not possess antimicrobial effects on Propionibacterium acnes and Streptococcus mutans. Membrane-depolarization assays and flow cytometric analyses showed that the antimicrobial properties of Glycinin-17, BCAS-16, and BCBS-11 against P. gingivalis, P. acnes, and S. mutans were dependent on membrane-disrupting potential. In contrast, major antimicrobial activities of these peptides against C. albicans were dependent on interactions with targets other than cell membranes. Furthermore, chromogenic Limulus amebocyte lysate assays showed that 50% effective concentrations (EC50, 0.12-0.31 mu M) of these three peptides neutralize LPS with similar potency (EC50: 0.11 mu M) to that of polymyxin B. Moreover, tube-formation assays in human umbilical vein endothelial cells showed similar angiogenic activities of the three peptides as that following treatment with LL-37. Although BCAS-16 exhibited hemolytic activity, the rate of hemolysis for Glycinin-17 and BCBS-11 in the presence of 500-mu M Glycinin-17 and BCBS-11 was less than 2%. These results demonstrate that cationic and amphipathic peptides from soybean proteins, particularly Glycinin-17 and BCBS-11, have potential as multifunctional ingredients for healthcare applications.

    DOI: 10.1002/bip.23023

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  • Platelet-rich fibrin prepared from stored whole-blood samples Reviewed

    Isobe Kazushige, Suzuki Masashi, Watanabe Taisuke, Kitamura Yutaka, Suzuki Taiji, Kawabata Hideo, Nakamura Masayuki, Okudera Toshimitsu, Okudera Hajime, Uematsu Kohya, Nakata Koh, Tanaka Takaaki, Kawase Tomoyuki

    INTERNATIONAL JOURNAL OF IMPLANT DENTISTRY   3   2017.3

  • Effects of arginine and leucine substitutions on anti-endotoxic activities andmechanisms of action of cationic and amphipathic antimicrobial octadecapeptide from rice alpha-amylase Reviewed

    Masayuki Taniguchi, Akihito Ochiai, Ryu Toyoda, Teppei Sato, Eiichi Saitoh, Tetsuo Kato, Takaaki Tanaka

    JOURNAL OF PEPTIDE SCIENCE   23 ( 3 )   252 - 260   2017.3

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    Previously, we showed that the antimicrobial cationic and amphipathic octadecapeptide AmyI-1-18 from rice a-amylase (AmyI-1) inhibited the endotoxic activity of lipopolysaccharide (LPS) from Escherichia coli. In addition, we demonstrated that several AmyI1-18 analogs containing arginine or leucine substitutions, which were designed on the basis of the helical wheel projection of AmyI-1-18, exhibited higher antimicrobial activity against human pathogenic microorganisms than AmyI-1-18. In the present study, anti-inflammatory (anti-endotoxic) activities of five AmyI-1-18 analogs containing arginine or leucine substitutions were investigated. Two single arginine-substituted and two single leucine-substituted AmyI-1-18 analogs inhibited the production of LPS-induced nitric oxide in mouse macrophages (RAW264) more effectively than AmyI-1-18. These data indicate that enhanced cationic and hydrophobic properties of AmyI-1-18 are associated with improved anti-endotoxic activity. In subsequent chromogenic Limulus amebocyte lysate assays, 50% inhibitory concentrations (IC50) of the three AmyI-1-18 analogs (G12R, D15R, and E9L) were 0.11-0.13 mu M, indicating higher anti-endotoxic activity than that of AmyI-1-18 (IC50, 0.22 mu M), and specific LPS binding activity. In agreement, surface plasmon resonance analyses confirmed direct LPS binding of three AmyI-1-18 analogs. In addition, AmyI-1-18 analogs exhibited little or no cytotoxic activity against RAW264 cells, indicating that enhancements of antiinflammatory and LPS-neutralizing activities following replacement of arginine or leucine did not result in significant increases in cytotoxicity. This study shows that the arginine-substituted and leucine-substituted AmyI-1-18 analogs with improved antiendotoxic and antimicrobial activities have clinical potential as dual-function host defense agents. Copyright (C) 2017 European Peptide Society and John Wiley & Sons, Ltd.

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  • Antimicrobial activity against <i>Porphyromonas gingivalis</i> and mechanism of action of the cationic octadecapeptide Amyl-1-18 and its amino acid-substituted analogs Reviewed

    M. Taniguchi, A. Ochiai, K. Takahashi, S. Nakamichi, T. Nomoto, E. Saitoh, T. Kato, T. Tanaka

    Journal of Bioscience and Bioengineering   122 ( 6 )   652 - 659   2016.12

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  • Preparation of a poly(L-lactic acid) membrane scaffold with open finger-like pores prepared by a nonsolvent-induced phase separation method with the aid of a surfactant Reviewed

    H. Minbu, T. Kawase, A. Ochiai, M. Taniguchi, T. Tanaka

    MEMBRANE   41 ( 6 )   304 - 310   2016.11

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    Here, we prepared a poly(L–lactic acid) (PLLA) microporous membrane with open finger–like pores as a scaffold for tissue engineering by a nonsolvent–induced phase separation method with the aid of a surfactant. The increase of surfactant content in the polymer solution caused the mold (glass plate) side of the membrane to adopt an indented structure with open finger–like pores to enhance the internal surface area. Osteoblast–like cells inoculated on the indented side grew on and in the PLLA membrane scaffold to reach 2 ~ 3 times higher cell density per unit apparent area compared to that attained in monolayer cultures. The cells on the membrane deposited calcium compounds by osteoinduction with ascorbic acid 2–phosphate, dexamethasone, and β– glycerophosphate. The PLLA membrane with open finger–like pores will likely be useful as scaffolds to support the implantation of osteogenic cells in bone tissue engineering.

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  • Rice Bran Protein as a Potent Source of Antimelanogenic Peptides with Tyrosinase Inhibitory Activity Reviewed

    Akihito Ochiai, Seiya Tanaka, Takaaki Tanaka, Masayuki Taniguchi

    JOURNAL OF NATURAL PRODUCTS   79 ( 10 )   2545 - 2551   2016.10

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    Rice (Oryza sativa) is consumed as a staple food globally, and rice bran, the byproduct, is an unused biomass that is ultimately discarded as waste. Thus, in the present study, a technique for producing tyrosinase inhibitory peptides from rice bran protein (RBP) was developed. Simultaneous treatment of RBP with chymotrypsin and trypsin produced numerous peptides. Subsequently, six tyrosinase inhibitory peptides were isolated from the hydrolysate fractions in a multistep purification protocol, and their amino acid sequences were determined. Three of these peptides had a C-terminal tyrosine residue and exhibited significant inhibitory effects against tyrosinase-mediated monophenolase reactions. Furthermore, peptide CT-2 (Leu-Gln-Pro-Ser-His-Tyr) potently inhibited melanogenesis in mouse B16 melanoma cells without causing cytotoxicity, suggesting the potential of CT-2 as an agent for melanin-related skin disorder treatment. The present data indicate that RBP is a potent source of tyrosinase inhibitory peptides and that simultaneous treatment of RBP with chymotrypsin and trypsin efficiently produces these peptides.

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  • AmyI-1-18, a cationic alpha-helical antimicrobial octadecapeptide derived from alpha-amylase in rice, inhibits the translation and folding processes in a protein synthesis system Reviewed

    Masayuki Taniguchi, Akihito Ochiai, Shun Fukuda, Teppei Sato, Eiichi Saitoh, Tetsuo Kato, Takaaki Tanakal

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   122 ( 4 )   385 - 392   2016.10

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    In our previous study, we used a cell-free rapid translation system (RTS), which is an in vitro protein synthesis system based on Escherichia coli lysate, for evaluating the inhibition of green fluorescent protein (GFP) synthesis by pyrrhocoricin. In this study, using an RTS, we evaluated the inhibition of GFP synthesis by AmyI-1-18, an antimicrobial octadecapeptide. We found that, similarly to pyrrhocoricin, Amyl-1-18 inhibited GFP synthesis in the RTS in a concentration-dependent manner. In addition, the blockage of transcription and translation steps in the RTS was individually estimated using RT-PCR after gene expression to determine the mRNA products and using sodium dodecyl sulfate-polyacrylamide gel electrophoresis to determine the amounts of GFP expressed from purified mRNA, respectively. The results demonstrated that the inhibition of GFP synthesis by AmyI-1-18 did not occur at the transcription step but rather at the translation step. Furthermore, we assessed the inhibition of DnaK-mediated refolding of chemically denatured luciferase by AmyI-1-18; Amyl-1-18 inhibited the protein folding activity of the ATP-dependent DnaK/DnaJ molecular chaperone system in a concentration-dependent manner. Surface plasmon resonance (SPR) analysis showed that Amyl-1-18 strongly bound to RNA with a ICD value of 1.4 x 10(-8) M but not to DNA and that Amyl-1-18 specifically bound to DnaK with a K-D value of 4.4 x 10(-8) M. These SPR analysis results supported the results obtained in both the RTS and the molecular chaperone system. These results demonstrated that both RNA and DnaK are most likely the target of AmyI-1-18 in the protein synthesis system. (C) 2016, The Society for Biotechnology, Japan. All rights reserved.

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  • New tyrosinase inhibitory decapeptide: Molecular insights into the role of tyrosine residues Reviewed

    Akihito Ochiai, Seiya Tanaka, Yuta Imai, Hisashi Yoshida, Takumi Kanaoka, Takaaki Tanaka, Masayuki Taniguchi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   121 ( 6 )   607 - 613   2016.6

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    Tyrosinase, a rate-limiting enzyme in melanin biosynthesis, catalyzes the hydroxylation of L-tyrosine to 3,4-dihydroxy-L-phenylalanine (L-dopa) (monophenolase reaction) and the subsequent oxidation of L-dopa to L-dopaquinone (diphenolase reaction). Thus, tyrosinase inhibitors have been proposed as skin-lightening agents; however, many of the existing inhibitors cannot be widely used in the cosmetic industry due to their high cytotoxicity and instability. On the other hand, some tyrosinase inhibitory peptides have been reported as safe. In this study, we found that the peptide TH10, which has a similar sequence to the characterized inhibitory peptide P4, strongly inhibits the monophenolase reaction with a half-maximal inhibitory concentration of 102 mu M. Seven of the ten amino acid residues in TH10 were identical to P4; however, TH10 possesses one N-terminal tyrosine, whereas P4 contains three tyrosine residues located at its N-terminus, center, and C-terminus. Subsequent analysis using sequence-shuffled variants indicated that the tyrosine residues located at the N-terminus and center of P4 have little to no contribution to its inhibitory activity. Furthermore, docking simulation analysis of these peptides with mushroom tyrosinase demonstrated that the active tyrosine residue was positioned close to copper ions, suggesting that TH10 and P4 bind to tyrosinase as a substrate analogue. (C) 2015 The Society for Biotechnology, Japan. All rights reserved.

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  • Pyrrhocoricin, a proline-rich antimicrobial peptide derived from insect, inhibits the translation process in the cell-free Escherichia coli protein synthesis system Reviewed

    Masayuki Taniguchi, Akihito Ochiai, Hiroshi Kondo, Shun Fukuda, Yohei Ishiyama, Eiichi Saitoh, Tetsuo Kato, Takaaki Tanaka

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   121 ( 5 )   591 - 598   2016.5

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    Previous studies have shown that pyrrhocoricin, a proline-rich antimicrobial peptide (PrAMP), killed sensitive species in a dose-dependent manner by specifically binding to DnaK. Here, on the basis of the finding that DnaK-deficient Escherichia coli strains are susceptible to PrAMPs, we used pyrrhocoricin to investigate internal targets other than DnaK. Using conventional antibiotics (bleomycin, streptomycin, and fosfomycin) that have known modes of action, first, we validated the availability of an assay using a cell-free rapid translation system (RTS), which is an in vitro protein synthesis system based on E. coli lysate, for evaluating inhibition of protein synthesis. We found that, similarly to bleomycin and streptomycin, pyrrhocoricin inhibited GFP synthesis in RTS in a concentration-dependent manner. In addition, blockage of transcription and translation steps in RTS was individually estimated using RT-PCR after gene expression to determine mRNA products and using sodium dodecyl sulfate-polyacrylamide gel electrophoresis to determine the amounts of GFP expressed from purified mRNA, respectively. The results demonstrated that this inhibition of GFP synthesis by pyrrhocoricin did not occur at the transcription step but rather at the translation step, in a manner similar to that of GFP synthesis by streptomycin, an inhibitor of the translation step by causing misreading of tRNA. These results suggest that RTS is a powerful assay system for determining if antimicrobial peptides inhibit protein synthesis and its transcription and/or translation steps. This is the first study to have shown that pyrrhocoricin inhibited protein synthesis by specifically repressing the translation step. (C) 2015, The Society for Biotechnology, Japan. All rights reserved.

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  • Effect of alanine, leucine, and arginine substitution on antimicrobial activity against candida albicans and action mechanism of a cationic octadecapeptide derived from -amylase of rice Reviewed

    Masayuki Taniguchi, Akihito Ochiai, Kiyoshi Takahashi, Shun-ichi Nakamichi, Takafumi Nomoto, Eiichi Saitoh, Tetsuo Kato, Takaaki Tanaka

    BIOPOLYMERS   106 ( 2 )   219 - 229   2016.3

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    AmyI-1-18, an antimicrobial peptide, is a cationic -helical octadecapeptide derived from -amylase of rice (Oryza sativa L. japonica) that contains four cationic amino acid residues (two arginines and two lysines). To enhance the antifungal activity of AmyI-1-18 against Candida albicans, 11 analogs bearing substitutions with alanine, leucine, and/or arginine, which were designed on the basis of the helical wheel projection of AmyI-1-18, were synthesized, and their antifungal activity was investigated. The antifungal activities of four analogs obtained by replacing arginine or lysine with alanine were significantly reduced. The results suggested that the cationic arginine and lysine residues in AmyI-1-18 are important for its antifungal activity. The antifungal activities of two single leucine-substituted analogs were not improved, but among three single arginine-substituted analogs, AmyI-1-18(D15R) had approximately a twofold higher antifungal activity [50% growth-inhibitory concentration (IC50): 31 M] than AmyI-1-18 (IC50: 64 M) and exhibited low hemolytic activity (4% at 100 M). Flow cytometric analysis using propidium iodide revealed that the antifungal activity of AmyI-1-18(D15R) was dependent on its membrane-disrupting activity in a manner different from that of AmyI-1-18. Further enhancement of the cationicity and hydrophobicity of AmyI-1-18(D15R) resulted in no improvement in antifungal activity and a significant increase in hemolytic activity. In this study, the results demonstrated that the antifungal activity of AmyI-1-18 against C. albicans was enhanced through increasing its membrane-disrupting activity by replacing aspartic acid at position 15 with arginine without a significant increase in hemolytic activity. (c) 2016 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 106: 219-229, 2016.

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  • Endotoxin-neutralizing activity and mechanism of action of a cationic alpha-helical antimicrobial octadecapeptide derived from alpha-amylase of rice Reviewed

    Masayuki Taniguchi, Akihito Ochiai, Kenta Matsushima, Koji Tajima, Tetsuo Kato, Eiichi Saitoh, Takaaki Tanaka

    PEPTIDES   75   101 - 108   2016.1

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    We have previously reported that AmyI-1-18, an octadecapeptide derived from alpha-amylase (AmyI-1) of rice, is a novel cationic alpha-helical peptide that exhibited antimicrobial activity against human pathogens, including Porphyromonas gingivalis, Pseudomonas aeruginosa, Prop ionibacterium acnes, Streptococcus mutans, and Candida albicans. In this study, to further investigate the potential functions of AmyI-1-18, we examined its inhibitory ability against the endotoxic activities of lipopolysaccharides (LPSs, smooth and Rc types) and lipid A from Escherichia coli. AmyI-1-18 inhibited the production of endotoxin-induced nitric oxide (NO), an inflammatory mediator, in mouse macrophages (RAW264) in a concentration-dependent manner. The results of a chromogenic Limulus amebocyte lysate assay illustrated that the ability [50% effective concentration (EC50): 0.17 mu M] of AmyI-1-18 to neutralize lipid A was similar to its ability (EC50: 0.26 mu M) to neutralize LPS, suggesting that AmyI-1-18 specifically binds to the lipid A moiety of LPS. Surface plasmon resonance analysis of the interaction between AmyI-1-18 and LPS or lipid A also suggested that AmyI-1-18 directly binds to the lipid A moiety of LPS because the dissociation constant (K-D) of AmyI-1-18 with lipid A is 5.6 x 10(-1)0 M, which is similar to that (4.3 x 10(-10) M) of AmyI-1-18 with LPS. In addition, AmyI-1-18 could block the binding of LPS-binding protein to LPS, although its ability was less than that of polymyxin B. These results suggest that AmyI-1-18 expressing antimicrobial and endotoxin-neutralizing activities is useful as a safe and potent host defense peptide against pathogenic Gram-negative bacteria in many fields of healthcare. (C) 2015 Elsevier Inc. All rights reserved.

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  • The heat-compression technique for the conversion of platelet-rich fibrin preparation to a barrier membrane with a reduced rate of biodegradation Reviewed

    Tomoyuki Kawase, Mana Kamiya, Mito Kobayashi, Takaaki Tanaka, Kazuhiro Okuda, Larry F. Wolff, Hiromasa Yoshie

    JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART B-APPLIED BIOMATERIALS   103 ( 4 )   825 - 831   2015.5

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    Platelet-rich fibrin (PRF) was developed as an advanced form of platelet-rich plasma to eliminate xenofactors, such as bovine thrombin, and it is mainly used as a source of growth factor for tissue regeneration. Furthermore, although a minor application, PRF in a compressed membrane-like form has also been used as a substitute for commercially available barrier membranes in guided-tissue regeneration (GTR) treatment. However, the PRF membrane is resorbed within 2 weeks or less at implantation sites; therefore, it can barely maintain sufficient space for bone regeneration. In this study, we developed and optimized a heat-compression technique and tested the feasibility of the resulting PRF membrane. Freshly prepared human PRF was first compressed with dry gauze and subsequently with a hot iron. Biodegradability was microscopically examined in vitro by treatment with plasmin at 37 degrees C or in vivo by subcutaneous implantation in nude mice. Compared with the control gauze-compressed PRF, the heat-compressed PRF appeared plasmin-resistant and remained stable for longer than 10 days in vitro. Additionally, in animal implantation studies, the heat-compressed PRF was observed at least for 3 weeks postimplantation in vivo whereas the control PRF was completely resorbed within 2 weeks. Therefore, these findings suggest that the heat-compression technique reduces the rate of biodegradation of the PRF membrane without sacrificing its biocompatibility and that the heat-compressed PRF membrane easily could be prepared at chair-side and applied as a barrier membrane in the GTR treatment. (c) 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 103B: 825-831, 2015.

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  • Quantitative single-cell motility analysis of platelet-rich plasma-treated endothelial cells in vitro Reviewed

    Tomoyuki Kawase, Takaaki Tanaka, Kazuhiro Okuda, Makoto Tsuchimochi, Masafumi Oda, Toshiaki Hara

    CYTOSKELETON   72 ( 5 )   246 - 255   2015.5

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    Platelet-rich plasma (PRP) has been widely applied in regenerative therapy due to its high concentration of growth factors. Previous in vitro and in vivo studies have provided evidence supporting the angiogenic activity of PRP. To more directly demonstrate how PRP acts on endothelial cells, we examined the PRP-induced changes in the motility of human umbilical vein endothelial cells by examining the involvement of VEGF. Time-lapse quantitative imaging demonstrated that in the initial phase (approximate to 2 h) of treatment, PRP substantially stimulated cell migration in a wound-healing assay. However, this effect of PRP was not sustained at significant levels beyond the initial phase. The average net distance of cell migration at 10 h was 0.45 +/- 0.16 mm and 0.82 +/- 0.23 mm in control and PRP-stimulated cells, respectively. This effect was also demonstrated with recombinant human VEGF and was significantly attenuated by a neutralizing anti-VEGF antibody. Immunofluorescent examination of paxillin and actin fibers demonstrated that PRP concomitantly up-regulated focal adhesion and cytoskeletal formation. Western blotting analysis of phosphorylated VEGFR2 demonstrated that PRP mainly stimulated the phosphorylation of immature VEGFR2 in a dose- and time-dependent manner, an action that was completely blocked by the neutralizing antibody. Taken together, these data suggest that PRP acts directly on endothelial cells via the activation of VEGFR2 to transiently up-regulate their motility. Thus, the possibility that PRP desensitizes target endothelial cells for a relatively long period of time after short-term activation should be considered when the controlled release system of PRP components is designed. (c) 2015 Wiley Periodicals, Inc.

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  • Preparation of poly(L-lactic acid) microfiltration membranes by a nonsolvent-induced phase separation method with the aid of surfactants Reviewed

    Hiromi Minbu, Akihito Ochiai, Tomoyuki Kawase, Masayuki Taniguchi, Douglas R. Lloyd, Takaaki Tanaka

    JOURNAL OF MEMBRANE SCIENCE   479   85 - 94   2015.4

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    Microfiltration membranes of poly(L-lactic acid) (PLEA) have been prepared by a nonsolvent-induced phase separation method with the aid of surfactants. Surfactants with hydrophilic-lipophilic balance (HLB) values of 14.9-15.6 were found to be useful in reducing the shrinkage in thickness of the PLEA membrane. Among the surfactants examined, Tween 80 was the best for preparing microfiltration membranes. The surfactant allowed instantaneous phase separation and seemed to enhance the diffusion of water in the PLEA solution during structure formation. The membrane had asymmetric finger-like structures and showed low membrane resistance and high bacterial cell retention when the membrane was prepared from a 10 wt% PLEA solution in 1,4-dioxane containing 10 wt.% Tween 80. Bovine serum albumin molecules passed through the membrane suggesting that the membrane functions as a microfiltration membrane. The membrane was stable at 25 degrees C but degradable at 60 degrees C in wet conditions. The membrane can be applied as a compostable microfiltration membrane in food and biochemical industries. (C) 2015 Elsevier B.V. All rights reserved.

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  • Antimicrobial Activity and Mechanism of Action of a Novel Cationic alpha-Helical Octadecapeptide Derived From alpha-Amylase of Rice Reviewed

    Masayuki Taniguchi, Akihito Ochiai, Kiyoshi Takahashi, Shun-ichi Nakamichi, Takafumi Nomoto, Eiichi Saitoh, Tetsuo Kato, Takaaki Tanaka

    BIOPOLYMERS   104 ( 2 )   73 - 83   2015.3

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    Amyl-1-18, an octadecapeptide derived from a-amylase (Amyl-1) of rice (Oryza sativa L. japonica), is a novel cationic a-helical antimicrobial peptide (AMP) that contains two lysine and two arginine residues. The antimicrobial activity of Amyl-1-18 against human pathogens was quantitatively evaluated using a chemiluminescence method that measures ATP derived from viable cells. Of the ten kinds of human pathogens, Amyl-1-18 exhibited antimicrobial activity against nine. Its 50% growth-inhibitory concentrations (ICs50) against Porphyromonas gingivalis, Propionibacterium acnes, Pseudomonas aeruginosa, Candida albicans, and Streptococcus mutans were 13, 19, 50, 64, and 77 mu M, respectively. Amyl-1-18 had little or no hemolytic activity even at 500 mu M, and showed negligible cytotoxicity up to 1200 mu M. The degree of 3,3'-dipropylthiadicarbocyanine iodide release from P. gingivalis cells induced by the addition of Amyl-1-18 was significantly lower than that induced by the addition of melittin. Flow cytometric analysis showed that the percentages of P. aeruginosa, S. mutans, and C. albicans cells stained with propidium iodide (PI), a DNA-intercalating dye, were 89%, 43%, and 3%, respectively, when Amyl-1-18 was added at a concentration equal to its 43IC(50). Therefore, the antimicrobial activity of Amyl-1-18 against P. aeruginosa and S. mutans appears to be mainly attributable to its membrane-disrupting activity. In contrast, its antimicrobial activity against P. gingivalis and C. albicans most likely depends upon interactions with intracellular targets other than their cell membranes. Collectively, these results indicate that Amyl-1-18 is useful as a safe and potent AMP against the pathogens described above in many fields of healthcare. (C) 2015 Wiley Periodicals, Inc.

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  • Contribution of Cationic Amino Acids toward the Inhibition of Arg-Specific Cysteine Proteinase (Arg-gingipain) by the Antimicrobial Dodecapeptide, CL(14-25), from Rice Protein Reviewed

    Masayuki Taniguchi, Yoshiyasu Matsuhashi, Takako K. Abe, Yohei Ishiyama, Eiichi Saitoh, Tetsuo Kato, Akihito Ochiai, Takaaki Tanaka

    BIOPOLYMERS   102 ( 5 )   379 - 389   2014.9

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    CL(14-25), a dodecapeptide, exhibits antimicrobial activity against Porphyromonas gingivalis with the 50% growth-inhibitory concentration (IC50) value of 145 mu M, and arginine-specific gingipain (Rgp)-inhibitory activity. Kinetic analysis revealed that CL(14-25) is a mixed-type inhibitor, with inhibition constants (K-i and K-i' values) of 1.4 x 10(-6) M and 4.3 x 10(-6) M, respectively. To elucidate the contributions of four cationic amino acid residues at the N- and C-termini of CL(14-25) toward Rgp-inhibitory activity, we investigated the Rgp-inhibitory activities of truncated and alanine-substituted analogs of CL(14-25). Rgp-inhibitory activities significantly decreased by truncated analogs, CL(15-25) and CL(1625), whereas those of CL(14-24) and CL(14-23) were almost as high as that of CL(14-25). Rgp-inhibitory activities of alanine-substituted analogs, CL(R14A) and CL(R14A, R15A) also significantly decreased, whereas those of CL(K25A) and CL(R24A, K25A) were higher than that of CL(14-25). These results suggest that the arginine residue at position 15 substantially contributes to the Rgp-inhibitory activity and that the arginine residue at position 14 plays important roles in exerting Rgp-inhibitory activity. In this study, we demonstrated that CL(K25A) was a potent, dual function, peptide inhibitor candidate, exhibiting Rgp-inhibitory activity with Ki and K-i(l) of 9.6 x 10(-7) M and 1.9 x 10(-6) M, respectively, and antimicrobial activity against P. gingivalis with an IC50 value of 51 mM. (C) 2014 Wiley Periodicals, Inc.

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  • An atmospheric-pressure plasma-treated titanium surface potentially supports initial cell adhesion, growth, and differentiation of cultured human prenatal-derived osteoblastic cells Reviewed

    Tomoyuki Kawase, Takaaki Tanaka, Hiromi Minbu, Mana Kamiya, Masafumi Oda, Toshiaki Hara

    JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART B-APPLIED BIOMATERIALS   102 ( 6 )   1289 - 1296   2014.8

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    An atmospheric-pressure plasma (APP) treatment was recently reported to render titanium (Ti) surfaces more suitable for osteoblastic cell proliferation and osteogenesis. However, the mechanism of action remains to be clearly demonstrated. In this study, we focused on cell adhesion and examined the effects of the APP treatment on the initial responses of human prenatal-derived osteoblastic cells incubated on chemically polished commercially pure Ti (CP-cpTi) plates. In the medium containing 1% fetal bovine serum, the initial cell adhesion and the actin polymerization were evaluated by scanning electron microscopy and fluorescence microscopy. The expression of cell adhesion-related molecules and osteoblast markers at the messenger RNA level was assessed by real-time quantitative polymerase chain reaction. Although the cells on the APP-treated CP-cpTi surface developed fewer cytoskeletal actin fibers, they attached with higher affinity and consequently proliferated more actively (1.46-fold over control at 72 h). However, most of the cell adhesion molecule genes were significantly downregulated (from 40 to 85% of control) in the cells incubated on the APP-treated CP-cpTi surface at 24 h. Similarly, the osteoblast marker genes were significantly downregulated (from 49 to 63% of control) at 72 h. However, the osteoblast marker genes were drastically upregulated (from 197 to 296% of control) in these cells by dexamethasone and beta-glycerophosphate treatment. These findings suggest that the APP treatment improves the ability of the CP-cpTi surface to support osteoblastic proliferation by enhancing the initial cell adhesion and supports osteoblastic differentiation when immature osteoblasts begin the differentiation process. (C) 2014 Wiley Periodicals, Inc.

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  • Crystal structure of alpha-amylase from Oryza sativa: molecular insights into enzyme activity and thermostability Reviewed

    Akihito Ochiai, Hiroshi Sugai, Kazuki Harada, Seiya Tanaka, Yohei Ishiyama, Kosuke Ito, Takaaki Tanaka, Toshio Uchiumi, Masayuki Taniguchi, Toshiaki Mitsui

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   78 ( 6 )   989 - 997   2014.6

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    AmyI-1 is an alpha-amylase from Oryza sativa (rice) and plays a crucial role in degrading starch in various tissues and at various growth stages. This enzyme is a glycoprotein with an N-glycosylated carbohydrate chain, a unique characteristic among plant alpha-amylases. In this study, we report the first crystal structure of AmyI-1 at 2.2-angstrom resolution. The structure consists of a typical (beta/alpha)(8)-barrel, which is well-conserved among most alpha-amylases in the glycoside hydrolase family-13. Structural superimposition indicated small variations in the catalytic domain and carbohydrate-binding sites between AmyI-1 and barley alpha-amylases. By contrast, regions around the N-linked glycosylation sites displayed lower conservation of amino acid residues, including Asn-263, Asn-265, Thr-307, Asn-342, Pro-373, and Ala-374 in AmyI-1, which are not conserved in barley alpha-amylases, suggesting that these residues may contribute to the construction of the structure of glycosylated AmyI-1. These results increase the depths of our understanding of the biological functions of AmyI-1.

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  • alpha-Amylase is a potential growth inhibitor of Porphyromonas gingivalis, a periodontal pathogenic bacterium Reviewed

    A. Ochiai, K. Harada, K. Hashimoto, K. Shibata, Y. Ishiyama, T. Mitsui, T. Tanaka, M. Taniguchi

    JOURNAL OF PERIODONTAL RESEARCH   49 ( 1 )   62 - 68   2014.2

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    Background and Objective: Porphyromonas gingivalis is a major etiological agent in the development and progression of periodontal diseases. In this study, we isolated a cell growth inhibitor against P. gingivalis species from rice protein extract.
    Material and Methods: The cell growth inhibitor active against P. gingivalis was purified from polished rice extract using a six-step column chromatography process. Its antimicrobial properties were investigated through microscope analysis, spectrum of activity and general structure.
    Results: The inhibitor was identified as AmyI-1, an alpha-amylase, and showed significant cell growth inhibitory activity against P. gingivalis species. Scanning electron microscopy micrograph analysis and bactericidal assay indicated an intriguing possibility that the inhibitor compromises the cell membrane structure of the bacterial cells and leads to cell death. Moreover, alpha-amylases from human saliva and porcine pancreas showed inhibitory activity similar to that of AmyI-1.
    Conclusions: This is the first study to report that alpha-amylases cause cell death of periodontal pathogenic bacteria. This finding highlights the potential importance and therapeutic potential of alpha-amylases in treating periodontal diseases.

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  • Effect of Substituting Arginine and Lysine with Alanine on Antimicrobial Activity and the Mechanism of Action of a Cationic Dodecapeptide (CL(14-25)), a Partial Sequence of Cyanate Lyase from Rice Reviewed

    Masayuki Taniguchi, Nobuteru Takahashi, Tomohiro Takayanagi, Atsuo Ikeda, Yohei Ishiyama, Eiichi Saitoh, Tetsuo Kato, Akihito Ochiai, Takaaki Tanaka

    BIOPOLYMERS   102 ( 1 )   58 - 68   2014.1

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    The antimicrobial activity of analogs obtained by substituting arginine and lysine in CL(14-25), a cationic alpha-helical dodecapeptide, with alanine against Porphyromonas gingivalis, a periodontal pathogen, varied significantly depending on the number and position of cationic amino acids. The alanine-substituted analogs had no hemolytic activity, even at a concentration of 1 mM. The antimicrobial activities of CL(K20A) and CL(K20A, K25A) were 3.8-fold and 9.1-fold higher, respectively, than that of CL(14-25). The antimicrobial activity of CL(R15A) was slightly lower than that of CL(14-25), suggesting that arginine at position 15 is not essential but is important for the antimicrobial activity. The experiments in which the alanine-substituted analogs bearing the replacement of arginine at position 24 and/or lysine at position 25 were used showed that arginine at position 24 was crucial for the antimicrobial activity whenever lysine at position 25 was substituted with alanine. Helical wheel projections of the alanine-substituted analogs indicate that the hydrophobicity in the vicinity of leucine at position 16 and alanines at positions 18 and/or 21 increased by substituting lysine at positions 20 and 25 with alanine, respectively. The degrees of diSC(3)-5 release from P. gingivalis cells and disruption of GUVs induced by the alanine-substituted analogs with different positive charges were not closely related to their antimicrobial activities. The enhanced antimicrobial activities of the alanine-substituted analogs appear to be mainly attributable to the changes in properties such as hydrophobicity and amphipathic propensity due to alanine substitution and not to their extents of positive charge (cationicity). (C) 2013 Wiley Periodicals, Inc.

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  • Antimicrobial activity and mechanism of action of a novel cationic alpha-helical dodecapeptide, a partial sequence of cyanate lyase from rice Reviewed

    Norihiro Takei, Nobuteru Takahashi, Tomohiro Takayanagi, Atsuo Ikeda, Kenji Hashimoto, Masahiro Takagi, Tsutomu Hamada, Eiichi Saitoh, Akihito Ochiai, Takaaki Tanaka, Masayuki Taniguchi

    PEPTIDES   42   55 - 62   2013.4

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    CL(14-25), a dodecapeptide, that is a partial region near N-terminus of cyanate lyase (CL, EC 4.3.99.1) from rice (Oryza sativa L. japonica), contains three arginine and two lysine residues. It was a novel cationic alpha-helical antimicrobial peptide. The antimicrobial activity of CL(14-25) against Porphyromonas gingivalis, a periodontal pathogen, was quantitatively evaluated by a chemiluminescence method that measures ATP derived from viable cells. The 50% growth-inhibitory concentration of CL(14-25) against P. gingivalis cells was 145 mu M. CL(14-25), even at a concentration of 800 mu M, had no hemolytic activity. When giant unilamellar vesicles (GUVs) that mimic the membrane composition of Gram-negative bacteria were used, microscopy image analysis suggested that CL(14-25) disrupted GUVs in a detergent-like manner. Therefore, CL(14-25) appears to exhibit antimicrobial activity through membrane disruption. To investigate the contribution of cationic amino acid residues in CL(14-25) to its antimicrobial activity, we synthesized four truncated CL analogs, in which one or two cationic amino acid residues were deleted from the N- and C- termini of CL(14-25). The degrees of calcein leakage from large unilamellar vesicles (LUVs) and 3,3'-dipropylthiadicarbocyanine iodide (diSC(3)-5) release from P. gingivalis cells induced by truncated CL analogs were closely related to their antimicrobial activities. CL analogs, which were truncated by removing an arginine residue from the N-terminus and a lysine residue from the C-terminus maintained their antimicrobial activity. However, CL analogs, which were further truncated by removing two arginine residues from the N-terminus, and an arginine and a lysine residue from the C-terminus, rarely exhibited antimicrobial activity. (c) 2012 Elsevier Inc. All rights reserved.

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  • Microporous membranes of PLLA/PCL blends for periosteal tissue scaffold Reviewed

    Tomoaki Kouya, Shin-ichiro Tada, Hiromi Minbu, Yu Nakajima, Makoto Horimizu, Tomoyuki Kawase, Douglas R. Lloyd, Takaaki Tanaka

    MATERIALS LETTERS   95   103 - 106   2013.3

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    Flexible microporous scaffold membranes of poly(L-lactic acid) (PLLA)/poly(epsilon-caprolactone) (PCL) blend were developed for bone regeneration. The new membranes overcome the fragility problems of PLLA membranes. When the PCL content in the blend was increased to 50%, the mechanical property for elongation was significantly improved without sacrificing the pores on the membrane surface that help tissue segments adhere the membrane. However, further increases in PCL content reduced the surface pores. In addition, spontaneous cell invasion and formation of cell-multilayers were observed in the membrane of PLLA/PCL blend (50:50). These results indicate that the porous membrane of PLLA/PCL blend (50:50) is useful for the preparation of periosteal sheets in tissue engineering. (C) 2012 Elsevier B.V. All rights reserved.

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  • Evaluation of positive interaction for cell growth between Bifidobacterium adolescentis and Propionibacterium freudenreichii using a co-cultivation system with two microfiltration modules Reviewed

    Tomoaki Kouya, Yohei Ishiyama, Takaaki Tanaka, Masayuki Taniguchi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   115 ( 2 )   189 - 192   2013.2

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    Using a co-cultivation system developed previously, positive interaction for cell growth between Bifidobacterium adolescent-is and Propionibacterium freudenreichii was evaluated. The total dry cell weight (DCW) of these two strains obtained in the co-cultivation system was 1.5-1.7-fold of the sum of the DCWs obtained in two single cultivations of each bacterium. (C) 2012, The Society for Biotechnology, Japan. All rights reserved.

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  • Antimicrobial activity and mechanism of action of a novel cationic β-helical octadecapeptide derived from heat shock protein 70 of rice Reviewed

    Masayuki Taniguchi, Atsuo Ikeda, Shun-Ichi Nakamichi, Yohei Ishiyama, Eiichi Saitoh, Tetsuo Kato, Akihito Ochiai, Takaaki Tanaka

    Peptides   48   147 - 155   2013

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    Hsp70(241-258), an octadecapeptide derived from the heat shock protein 70 (Hsp70) of rice (Oryza sativa L. japonica), is a novel cationic β-helical antimicrobial peptide (AMP) that contains four lysine, two arginine, and two histidine residues. The antimicrobial activity of Hsp70(241-258) against Porphyromonas gingivalis, a periodontal pathogen, and Candida albicans, an opportunistic fungal pathogen, was quantitatively evaluated using a chemiluminescence method that measures ATP derived from viable cells. The 50% growth-inhibitory concentrations of Hsp70(241-258) against P. gingivalis and C. albicans cells were 63μM and 70μM, respectively. Hsp70(241-258) had little or no hemolytic activity even at 1 mM, and showed negligible cytotoxicity up to 300M. The degrees of calcein leakage from large unilamellar vesicles, which mimic the membranes of Gram-negative bacteria, and 3,3 β- dipropylthiadicarbocyanine iodide release from P. gingivalis cells induced by the addition of Hsp70(241-258) increased in a concentrationdependent manner. When Hsp70(241-258) was added to calcein-acetoxymethyl ester-loaded C. albicans cells, calcein release from the cells increased in a concentration-dependent manner. Flow cytometric analysis also showed that the percentages of C. albicans cells stained with propidium iodide, a DNAintercalating dye, increased as the concentration of Hsp70(241-258) added was increased. Therefore, Hsp70(241-258) appears to exhibit antimicrobial activity against P. gingivalis and C. albicans through membrane disruption. These results suggest that Hsp70(241-258) could be useful as a safe and potent AMP against P. gingivalis and C. albicans in many fields of health care, especially in the control of oral infections. © 2013 Elsevier Inc. All rights reserved.

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  • Evaluation of positive interactions for cell growth between bifidobacterial strains and propionibacterium freudenreichii by using a co-cultivation system Reviewed

    Tomoaki Kouya, Yohei Ishiyama, Takaaki Tanaka, Masayuki Taniguchi

    Journal of Chemical Engineering of Japan   46 ( 3 )   226 - 229   2013

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    Positive interactions between Bifidobacterium longum or Bifidobacterium breve and Propionibacterium freudenreichii that enhance cell growth were evaluated using a co-cultivation system. The total dry cell weight of B. longum and P. freudenreichii from co-cultivation was 10.6 g. This value is 1.2-fold higher than the sum (9.1 g) of the maximum dry cell weights obtained from the cultivation of each bacterium. Currently, the total dry cell weight of co-cultivated B. breve and P. freudenreichii was 21.3 g. The total dry cell weight of B. breve and P. freudenreichii is 1.6-fold higher than the sum (13.2 g) of the maximum dry cell weights from two single cultivations of each bacterium. These data are direct evidence for a cooperative interaction between the two microorganisms. Copyright © 2013 The Society of Chemical Engineers, Japan.

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  • Formation of depth filter microfiltration membranes of poly(L-lactic acid) via phase separation Reviewed

    Takaaki Tanaka, Takayuki Nishimoto, Kazuhiro Tsukamoto, Masaharu Yoshida, Tomoaki Kouya, Masayuki Taniguchi, Douglas R. Lloyd

    JOURNAL OF MEMBRANE SCIENCE   396   101 - 109   2012.4

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    Depth filter microfiltration membranes produced from biodegradable poly(l-lactic acid) (PLLA) are reported in this paper. The asymmetric porous membranes reported here were formed via a process that combined nonsolvent-induced and thermally induced phase separation. The membranes served as efficient and rapid depth filters to retain bacterial cells when cell suspensions were filtered from the porous or rough side of the membrane, while serving as screen filters when the suspension was fed from the smooth skin side. After use, the depth filters were disposed of by non-enzymatic degradation - a process that was significantly accelerated in wet conditions at 60 degrees C, conditions under which composting is usually operated. The fact that the PLLA depth filters reported here can be disposed of by composting will help reduce industrial wastes in biochemical and food industries when they are used as a pre-filter. (C) 2012 Elsevier B. V. All rights reserved.

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  • An osteogenic grafting complex combining human periosteal sheets with a porous poly(L-lactic acid) membrane scaffold: Biocompatibility, biodegradability, and cell fate in vivo Reviewed

    Tomoyuki Kawase, Takaaki Tanaka, Takayuki Nishimoto, Kazuhiro Okuda, Masaki Nagata, Douglas M. Burns, Hiromasa Yoshie

    JOURNAL OF BIOACTIVE AND COMPATIBLE POLYMERS   27 ( 2 )   107 - 121   2012.3

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    In this in vitro study, novel porous poly(L-lactic acid) membranes were developed to improve periosteal sheets by promoting initial adhesion of periosteal tissue segments and stimulating the formation of a viable multilayered cellular sheet. The biocompatibility, biodegradability, and osteogenicity were evaluated using human periosteal tissue segments cultured on porous poly(L-lactic acid) membranes; the periosteal sheets were osteogen induced and were then implanted in the dorsal subcutaneous tissue of nude mice. In vivo, the membrane degraded into clusters of membrane particles separated by wide cracks; fibroblastic cells invaded along with small blood vessels from the surrounding mouse connective tissue. In osteoinduced periosteal sheets, the membrane clusters were surrounded by numerous capillaries and a number of tartrate-resistant acid phosphatase-positive, multinucleated cells. Neither severe inflammation nor fibrous encapsulation was observed throughout the implantation (similar to 12 weeks). These porous poly(L-lactic acid) membranes were highly biocompatible and functioned well as biodegradable scaffolds that could enhance the use of osteogenic periosteal sheets in therapy.

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  • Depth Filtration of Cell Suspensions of Agglomerated Streptobacteria by Biodegradable Membranes Reviewed

    Masaharu Yoshida, Masayuki Taniguchi, Takaaki Tanaka

    JOURNAL OF CHEMICAL ENGINEERING OF JAPAN   45 ( 9 )   778 - 784   2012

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    Filtration is an important industrial production processes to treat particle dispersion systems. In this paper we show the filtration characteristics of suspensions of agglomerated streptobacteria (chain bacteria), Bacillus amyloliquefaciens, by asymmetric depth filter membranes of poly(L-lactic acid) (PLLA). The permeation flux was higher in the filtration with the depth filter membrane than in that with a screen filter membrane at a transmembrane pressure of 50-150 kPa. The bacterial cell layer had a high compressibility index of the specific resistance (0.60) in the filtration with the screen filter membrane. However, the cell layer formed in the internal structure of the depth filter membrane of PLLA showed a low apparent compressibility index (0.11). Scanning electron micrographs revealed that the agglomerated cells were captured on the depth filter membrane at 10 kPa while those cells were captured within the membrane at 150 kPa. Thus the filtration performance with the depth filter membrane increased at higher transmembrane pressures. The biodegradable PLLA membrane showed high performance in the filtration of bacterial cell suspensions and has an advantage in disposal after use because it can be compostable with bacterial cells clogged in the membrane.

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  • Improved adhesion of human cultured periosteal sheets to a porous poly(L-lactic acid) membrane scaffold without the aid of exogenous adhesion biomolecules Reviewed

    Tomoyuki Kawase, Takaaki Tanaka, Takayuki Nishimoto, Kazuhiro Okuda, Masaki Nagata, Douglas M. Burns, Hiromasa Yoshie

    JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A   98A ( 1 )   100 - 113   2011.7

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    Human cultured periosteal sheets, which are developed from small excised periosteum tissue segments (PTSs) in culture dishes by simple expansion culture, have been applied as a promising autologous osteogenic grafting material for periodontal regenerative therapy. However, the weak initial adhesion of PTSs to dish surfaces often hampers cellular outgrowth and limits the number of preparations. To correct this weakness and still avoid the use of animal-derived adhesion biomolecules, we have developed a novel, biodegradable, porous poly(L-lactic acid) (pPLLA) membrane. Freshly excised PTSs bound well to the highly porous pPLLA membrane, possibly due to the presence of semihemispheric 20-30 mu m diameter openings on the upper surface. Global gene expression analysis demonstrated that periosteal sheets cultured on pPLLA membranes upregulated expression of many adhesion molecules. Osteogenic induction stimulated the production of proteoglycans by these cells and concomitantly enhanced their expansion and penetration into the deep pore regions of the membrane in parallel with the progression of in vitro mineralization. These findings suggest that our pPLLA membranes not only facilitate initial adhesion, primarily mediated by adsorbed proteins, but also enhance biological adhesion by inducing endogenous adhesion molecules in periosteal sheet cultures. Therefore, the efficacy of periosteal sheets in therapy should be greatly enhanced by using this new pPLLA membrane. (C) 2011 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 98A: 100-113, 2011.

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  • Poly(L-lactic acid) Microfiltration Membrane Formation via Thermally Induced Phase Separation with Drying Reviewed

    Takaaki Tanaka, Masatou Ueno, Youhei Watanabe, Tomoaki Kouya, Masayuki Taniguchi, Douglas R. Lloyd

    JOURNAL OF CHEMICAL ENGINEERING OF JAPAN   44 ( 7 )   467 - 475   2011

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    Microfiltration membranes of poly(L-lactic acid) (PLLA), which is a major biodegradable plastic, were prepared from PLLA-1,4-dioxane-water solutions via the thermally induced phase separation. The resulting membranes are used to retain bacterial cells and permeate protein molecules. Only 20% of the bacterial cells in cell suspensions were retained by the membranes prepared from 10-12.5% polymer solution. Increasing the polymer concentration from 12.5 to 15% markedly elevated the bacterial cell retention to 60%. The retention further increased to 90% by elevating the casting temperature from 40 to 50 degrees C. By drying the polymer solution for 2 min before quenching, the cell retention reached 99%. Casting at temperatures of 60 degrees C or higher and drying for periods of 5 min or longer made the membrane uneven and/or increased the membrane resistance. By drying the polymer solution before quenching, the porous structure near the membrane surface was altered so that the membrane retains the bacterial cells. The membrane retained bacterial cells in the manner of a screen filter. The membranes developed in this research will be useful as a pre-filter in biochemical processes to enhance the life time of the final filters and to reduce industrial wastes due to being made of a compostable polymer.

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  • Growth-inhibition of hiochi bacteria in namazake (raw sake) by bacteriocins from lactic acid bacteria Reviewed

    Masayuki Taniguchi, Yohei Ishiyama, Takeomi Takata, Toshihiro Nakanishi, Mitsuoki Kaneoke, Ken-ichi Watanabe, Fujitoshi Yanagida, Yi-sheng Chen, Tomoaki Kouya, Takaaki Tanaka

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   109 ( 6 )   570 - 575   2010.6

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    The bacteriocins produced by Lactococcus lactis subsp. lactis C101910 (C101910) and NBRC 12007 (NBRC 12007) were used to prevent the growth of sake spoiling hiochi bacteria (Lactobacillus hilgardii, Lactobacillus fructivorans, and Lactobacillus paracasei) in namazake, which is raw (unpasteurized) sake. The bacteriocin concentrations required for decreasing the viable cell concentrations of L hilgardii and L fructivorans below the detection limit (1.0 x 10(2) cells/ml) in 24 h from the initial concentration of 4.0-9.5 x 10(5) cells/ml in the namazake at pH 4.5 and at 4 degrees C, were 18-35 U/ml and 5.6 U/ml for the bacteriocin from C101910 and NBRC 12007, respectively. To decrease the viable cell concentration of L paracasei from the initial concentration of 7.5 x 10(5) cells/ml to below the detection limit (1.0 x 10(2) cells/ml) in 24 h, 350 U/ml bacteriocin from C101910 and 140 U/ml bacteriocin from NBRC 12007 were required. In experiments using McIlvaine buffer (pH 4.5) with 15% ethanol instead of namazake as the medium, the viable cell concentrations of L. hilgardii and L paracasei decreased to less than 1.0 x 10(2) cells/ml, whereas those of L fructivorans decreased to less than 1.0 x 10(3) cells/ml, when bacteriocins were added at the concentrations that had proven effective in namazake. The membrane depolarization assay using a fluorescent probe showed that the presence of ethanol stimulated the collapse of the membrane potential induced by bacteriocins. The ethanol induced collapse of the membrane potential suggests that the application of bacteriocins at the storage stage of namazake is more beneficial than when used in other stages of the sake brewing process. (C) 2009, The Society for Biotechnology, Japan. All rights reserved.

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  • Mechanical properties of microporous foams of biodegradable plastic Reviewed

    Takaaki Tanaka, Takashi Aoki, Tomoaki Kouya, Masayuki Taniguchi, Wataru Ogawa, Yuuji Tanabe, Douglas R. Lloyd

    DESALINATION AND WATER TREATMENT   17 ( 1-3 )   37 - 44   2010.5

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    Permeable microporous foams and membranes of biodegradable polyesters are currently used in the area of tissue engineering and drug delivery systems. Their mechanical properties are useful in the medical applications. The foams should mechanically fit to the tissues where the foams were implanted. In this study, we investigated the mechanical properties of the foams of biodegradable polyesters by compression tests. Microporous foams of poly(L-lactic acid), poly (epsilon-caprolactone), and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) were prepared by thermally induced phase separation method. The compression tests were performed at 23-24 degrees C with a universal testing machine. The structure of the microporous foams depended on the kinds of polymers, polymer concentrations, and quenching temperatures. The cell size of the foams was smaller when the polymer concentration was higher or the quenching temperature was lower. We analyzed the stress-strain diagrams of the foams in the compression test. The lower the relative density of the foams to the solid materials the lower the elastic limit stress was. The relative Young&apos;s modulus and relative elastic limit stress of the foams were approximately proportional to the square of their relative density and less dependent on their cell size. The dependences were similar to those of open-cell foams of polyurethane.

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  • Filtration characteristics of carbon nanotubes and preparation of buckypapers Reviewed

    Takaaki Tanaka

    DESALINATION AND WATER TREATMENT   17 ( 1-3 )   193 - 198   2010.5

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    Membrane filtration is often used in purification of carbon nanotubes and in preparation of their thin films (buckypaper). Filtration characteristics of multiwall carbon nanotubes (MWCNT) suspended in aqueous solution of detergents was investigated in this study. Although the diameter of the MWCNT was estimated at around 50 nm from the scanning electron micrograph, a microfiltration membrane whose nominal pore size was 0.2 mu m retained the carbon nanotubes. The specific resistance of the filter cake of the carbon nanotubes was 7 x 10(13) m kg(-1) at 98 kPa and the compressibility index was 0.12. The carbon nanotubes were entangled in the buckypaper formed on the microfiltration membrane and the internal structure of the buckypaper was similar under the applied pressure in the preparation of 10-98 kPa.

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  • Formation of microporous membranes of poly(1,4-butylene succinate) via nonsolvent and thermally induced phase separation Reviewed

    Takaaki Tanaka, Masaki Takahashi, Shigeko Kawaguchi, Takeru Hashimoto, Hiroshi Saitoh, Tomoaki Kouya, Masayuki Taniguchi, Douglas R. Lloyd

    DESALINATION AND WATER TREATMENT   17 ( 1-3 )   176 - 182   2010.5

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    Microporous membranes of poly(1,4-butylene succinate) (PBS), which is a biodegradable biomass plastic, were prepared from PBS-chloroform solutions via nonsolvent and thermally induced phase separation with a coagulation bath of methanol. The permeation resistance of a membrane prepared from a 10% polymer solution at 25 degrees C was more than 10(14) m(-1). The membrane resistance decreased with increasing pre-incubation temperature. Retention of bacterial cells (0.7 phi x 2.5 mu m) decreased with increasing pre-incubation temperature. Increasing polymer concentration increased the permeation resistance and the retention of the cells. Membranes prepared from the 10% PBS solution pre-incubated at 47.5 degrees C show a permeation resistance of 10(11) m(-1) and retention of the bacterial cells of 99%. The PBS membranes will be useful in biosepartion processes as a prefilter that can be disposed by composting after use.

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  • Enzymatic oxidative polymerization of methoxyphenols Reviewed

    Takaaki Tanaka, Masafumi Takahashi, Hiroyuki Hagino, Shin-Ichi Nudejima, Hisaki Usui, Tomoyuki Fujii, Masayuki Taniguchi

    CHEMICAL ENGINEERING SCIENCE   65 ( 1 )   569 - 573   2010.1

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    Enzymatic oxidative polymerization of methoxyphenols by a fungal laccase was investigated to synthesize polymethoxyphenols, which would be environmentally benign materials because they are degraded by lignin degrading fungi. An industrially produced thermostable laccase from Trametes sp. Ha1 polymerized o-methoxyphenol and 2,6-dimethoxyphenol in McIlvaine buffer (pH 4.5) both with and without organic solvents while the enzyme needed organic solvents to polymerize phenol, alkylphenols, and m- and p-methoxyphenols. The reaction rates correlated with the energies of the highest occupied molecular orbitals (HOMO) of the phenols. The poly(o-methoxyphenol) synthesized was soluble in N,N-dimethylformamide (DMF) while 80% of the poly(2,6-dimethoxyphenol) was insoluble in DMF. The weight-average molecular weight of the poly(o-methoxyphenol) was 7000-11,000. The reaction rate of polymerization in McIlvaine buffer without organic solvents increased until an o-methoxyphenol concentration of 500 mmol dm(-3) although the Michaelis constant calculated from the oxygen consumption rate was 0.4 mmol dm(-3). The o-methoxyphenol and 2,6-dimethoxyphenol would be suitable for production of environmentally benign phenolic resins by enzymatic oxidative polymerization using laccase without formaldehyde, hydrogen peroxide, or organic solvent. (C) 2009 Elsevier Ltd. All rights reserved.

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  • Effect of steam explosion pretreatment on treatment with Pleurotus ostreatus for enzymatic hydrolysis of rice straw. Reviewed

    M. Taniguchi, D. Takahashi, D. Watanabe, K. Sakai, K. Hoshino, T. Kouya, T. Tanaka

    J. Biosci. Bioeng.   110 ( 4 )   449 - 452   2010

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    The effects of steam explosion (1.5MPa, 1min) on the treatment of rice straw with Pleurotus ostreatus were evaluated in terms of the change in composition of the components and the susceptibility to enzymatic hydrolysis. When rice straw was pretreated with a steam explosion prior to biological treatment, the treatment time required for obtaining a 33% net glucose yield was reduced to 36 days from 60 days. The reduction is probably due to loosening of networks of Klason lignin with sugar moieties and partial collapse of the structure during the biological treatment.

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  • Microfiltration of Bacterial Cell Suspensions with an Asymmetric Depth Filter Reviewed

    Masaharu Yoshida, Masayuki Taniguchi, Takaaki Tanaka

    KAGAKU KOGAKU RONBUNSHU   36 ( 5 )   494 - 500   2010

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    Filtration characteristics of an asymmetric depth filter (nominal pore size: 0.2 mu m) were investigated by using suspensions of two bacteria, Lactobacillus plantarum and Bacillus amyloliquefaciens. In the filtration of L. plantarum suspensions, the permeation flux was 3-8 times higher with the depth filter than with a screen filter at 10-150 kPa. Apparent specific resistance of the cell layer in the depth filter reduced to about 1/10-1/100 of that of the filter cake formed on the screen filter. Scanning electron microscopy revealed that the trapped microbial cells were dispersed to a depth of 2/3 from the inlet side of the depth filter. In the filtration of B. amyloliquefaciens suspensions, the permeation flux of depth and screen filters was comparable at 10-50 kPa, and filter cakes formed on both filters. However, the permeation flux increased remarkably at 100-150 kPa with the depth filter but scarcely increased with the screen filter. The smallness of increase with the screen filter was due to the high compressibility of the microbial filter cake, while the remarkable increase with the depth filter was due to the penetration of the chain bacterium into the filter. The increase in transmembrane pressure was highly effective in the filtration of agglomerated chain bacteria such as B. amyloliquefaciens in trapping cells in the asymmetric depth filter.

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  • Evaluation of Fungal Pretreatments for Enzymatic Saccharification of Rice Straw Reviewed

    Masayuki Taniguchi, Daisuke Takahashi, Daisuke Watanabe, Kenji Sakai, Kazuhiro Hoshino, Tomoaki Kouya, Takaaki Tanaka

    JOURNAL OF CHEMICAL ENGINEERING OF JAPAN   43 ( 4 )   401 - 405   2010

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    To enzymatically hydrolyze cellulose of rice straw to glucose, the effect of pretreatment of rice straw with 15 strains of basidiomycetes is evaluated in terms of the quantitative changes in the components of pretreated rice straw and their susceptibility to enzymatic hydrolysis. Pleurotus ostreatus was found to be one of the most suitable white-rot fungi for biological pretreatment of rice straw. Of 11 strains of P. ostreatus tested, ATCC 66376 was found to have the properties superior to the other strains. Thus, in the pretreatment for 48 d, P. ostreatus ATCC 66376 degraded 39% Klason lignin and retained 79% cellulose in the pretreated rice straw. When the rice straw pretreated with this fungus was hydrolyzed with a commercial cellulase, the net yield of glucose determined on the basis of the weight of cellulose fraction of the untreated rice straw was the highest.

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  • 相分離法を用いたポリ乳酸多孔質膜の作製

    上野 正統, 渡辺 洋平, 高屋 朋彰, 谷口 正之, 田中 孝明

    化学工学会 研究発表講演要旨集   2010   1072 - 1072   2010

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    DOI: 10.11491/scej.2010f.0.1072.0

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  • ポリ乳酸製デプスフィルターの開発

    西本 崇之, 吉田 政晴, 高屋 朋彰, 谷口 正之, 田中 孝明

    化学工学会 研究発表講演要旨集   2010   716 - 716   2010

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    DOI: 10.11491/scej.2010f.0.716.0

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  • 非対称型デプスフィルターによる細菌懸濁液の精密濾過

    吉田 政晴, 谷口 正之, 田中 孝明

    化学工学会 研究発表講演要旨集   2010   715 - 715   2010

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    DOI: 10.11491/scej.2010f.0.715.0

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  • ポリ乳酸-ヒドロキシアパタイト-複合膜

    多田 晋一郎, 高屋 朋彰, 谷口 正之, 田中 孝明

    化学工学会 研究発表講演要旨集   2010   971 - 971   2010

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    DOI: 10.11491/scej.2010f.0.971.0

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  • ラッカーゼを用いたポリフェノールバイオセンサーの開発

    豊島 永祥, 高屋 朋彰, 谷口 正之, 田中 孝明

    化学工学会 研究発表講演要旨集   2010   998 - 998   2010

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    DOI: 10.11491/scej.2010f.0.998.0

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  • Depth filter membranes of biodegradable biomass plastics

    Takaaki Tanaka, Takayuki Nishimoto, Tomoaki Kouya, Masayuki Taniguchi, Douglas R. Lloyd

    AIChE Annual Meeting, Conference Proceedings   2010

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    Depth filter membranes of conventional synthetic polymers have problems at disposal after their uses although they can avoid the formation of filter cakes which cause a large increase in membrane resistance. In this study we developed microfiltration membranes of poly(L-lactic acid), one of the biodegradable biomass plastics. The asymmetric porous structures of the membranes were formed via thermally and nonsolvent-induced phase separation process. The membranes retained bacterial cells in a manner of depth filters. Depth filter membranes of biodegradable biomass plastics can be degraded in composting machines to reduce industrial wastes in biochemical and food industries.

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  • Growth-inhibitory activity of antimicrobial peptides from rice against oral pathogenic bacteria and clarification of their action mechanism Reviewed

    Tomoaki Kouya, Yusuke Koiso, Mayumi Taiyouji, Sadami Ohtsubo, Takaaki Tanaka, Masayuki Taniguchi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   108   S141 - S141   2009.11

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  • Growth-inhibition of Lactobacillus hilgardii, a bacterium related to hiochi, in the mizu-koji process by bacteriocins from lactic acid bacteria. Reviewed

    Y. Ishiyama, T. Takata, T. Nakanishi, M. Kaneoke, K. Watanabe, F. Yanagida, Y. Chen, T. Kouya, T. Tanaka, M. Taniguchi

    Jpn. J. Food Eng.   10 ( 1 )   45 - 53   2009

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  • Microfiltration of Incompressible Particle Suspension with an Asymmetric Depth Filter Reviewed

    Takaaki Tanaka, Tomoya Mimura, Mitsuo Koga, Masaharu Yoshida, Masayuki Taniguchi, Kazuhiro Nakanishi

    KAGAKU KOGAKU RONBUNSHU   35 ( 1 )   81 - 86   2009

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    Depth filters trap particles internally, and thus the compaction of the particle layer is partially repressed by the internal structure of the filter to yield a high permeation flux at a low transmembrane pressure. We filtered suspensions of three kinds of polystyrene latex particles (incompressible, spherical, 0.30, 0.54, and 0.70 mu m in diameter) with an asymmetric depth filter, SE20 (nominal pore size=0.20 mu m), at different transmembrane pressures and concentrations. The filtration resistance per unit weight of trapped particles with the depth filter was half of that with a screen filter in the filtration of latex particles. The particles were trapped first near the outlet side of the filter and then near the inlet side, unlike the standard blocking model for the membranes with cylindrical pores and the deep bed filtration model for sand filtration. A filtration model was proposed for the filtration of latex suspension with the depth filter. The model suggested that the asymmetric depth filter showed high performance because the filaments in the filter increased the porosity of the particle layer in contact with them.

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  • Enzymatic treatment of estrogens and estrogen glucuronide Reviewed

    Takaaki Tanaka, Toshiyuki Tamura, Yuuichi Ishizaki, Akito Kawasaki, Tomokazu Kawase, Masahiro Teraguchi, Masayuki Taniguchi

    JOURNAL OF ENVIRONMENTAL SCIENCES   21 ( 6 )   731 - 735   2009

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    Natural and synthetic estrogens from sewage treatment systems are suspected to influence the reproductive health of the animals in the rivers. In this article, we investigated the enzymatic treatment of three estrogens (estrone, 17 beta-estradiol, and 17 alpha-ethynyletstradiol) by a fungal laccase which oxidize phenolic compounds with dissolved oxygen. The elimination of the estrogenic activities by enzymatic oxidation was demonstrated by medaka vitellogenin assay. In addition, we developed an enzymatic treatment system comprised of beta-D-glucuronidase and the laccase for 17 beta-estradiol 3-(beta-D-glucuronide) degradation. The two enzymes eliminated 17 beta-estradiol 3-(beta-D-glucuronide) and the intermediate, 17 beta-estradiol, efficiently.

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  • Microporous foams of polymer blends of poly(L-lactic acid) and poly(epsilon-caprolactone) Reviewed

    Takaaki Tanaka, Satoko Eguchi, Hiroshi Saitoh, Masayuki Taniguchi, Douglas R. Lloyd

    DESALINATION   234 ( 1-3 )   175 - 183   2008.12

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    Microporous foams of polymer blends of two biodegradable polyesters, poly(L-lactic acid) (PLLA) and poly(E-caprolactone) (PCL), were prepared via thermally induced phase separation method. The phase behaviors of the solutions of the polymer blends in 1,4-dioxane containing water were similar, which would be due to the similar Solubility parameters of the two polymers (20.5-21.1 and 21.2 MPa(0.5)),). However, the porous structure of the polymer blends dependent oil the blend ratio under the same conditions other than the ratio. Hepatic cells (HepG2 cells) were Cultured on the porous polymer blends. The cells invaded into the scaffold of PCL and PLLA-PCL-blend (1:4) by 1.0 and 1.5 mm, respectively, while they grew near the Surface of the PLLA foams. Blending of biodegradable polyesters has the potential of controlling the porous structure of biodegradable foams.

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  • Production of bacteriocin by Staphylococcus sp. NPSI 38 in koji extract medium with rice protein hydrolyzate and its growth-inhibitory activity against hiochi-bacteria Reviewed

    Yohei Ishiyama, Takeomi Takata, Toshihiro Nakanishi, Naomi Watanabe, Mistuoki Kaneoke, Ken-Ichi Watanabe, Takaaki Tanaka, Masayuki Taniguchi

    Food Science and Technology Research   14 ( 3 )   239 - 248   2008.5

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    Enhanced production of bacteriocin by Staphylococcus sp. NPSI 38 (NPSI 38) using koji extract medium was investigated. The bacteriocin produced by NPSI 38 in MRS medium was found to be effective for inhibiting the growth of Lactobacillus hilgardii NBRC 15886T, one of the representative hiochi-bacteria. In cultivations using test tubes without pH control, meat extract and Polypepton were effective for bacteriocin production by NPSI 38 with 10%(v/v) koji extract medium. When the koji extract medium supplemented with hydrolyzate of a rice protein preparation was used instead of meat extract and Polypepton, NPSI 38 produced 160 U/ml of bacteriocin in the cultivation with pH control, which was almost as high as that (156 U/ml) observed in cultivation using MRS medium with pH control. When the cells of L. hilgardii NBRC 15886 T collected at logarithmic growth and stationary phases were inoculated into fresh modified MRS medium with 11 U/ml of the bacteriocin, negligible cell growth was observed, irrespective of different growth phases of cells inoculated. Little or no increase in cell concentrations in the media containing bacteriocin showed that the action of the bacteriocin produced by NPSI 38 was bacteriostatic against the hiochi-bacterium.

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  • Production of bacteriocin by Staphylococcus sp NPSI 38 in koji extract medium with rice protein hydrolyzate and its growth-inhibitory activity against hiochi-bacteria Reviewed

    Yohei Ishiyama, Takeomi Takata, Toshihiro Nakanishi, Naomi Watanabe, Mistuoki Kaneoke, Ken-ichi Watanabe, Takaaki Tanaka, Masayuki Taniguchi

    FOOD SCIENCE AND TECHNOLOGY RESEARCH   14 ( 3 )   239 - 248   2008.5

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    Enhanced production of bacteriocin by Staphylococcus sp. NPSI 38 (NPSI 38) using koji extract medium was investigated. The bacteriocin produced by NPSI 38 in MRS medium was found to be effective for inhibiting the growth of Lactobacillus hilgardii NBRC 15886(T), one of the representative hiochi-bacteria. In cultivations using test tubes without pH control, meat extract and Polypepton were effective for bacteriocin production by NPSI 38 with 10%(v/v) koji extract medium. When the koji extract medium supplemented with hydrolyzate of a rice protein preparation was used instead of meat extract and Polypepton, NPSI 38 produced 160 U/ml of bacteriocin in the cultivation with pH control, which was almost as high as that (156 U/ml) observed in cultivation using MRS medium with pH control. When the cells of L. hilgardii NBRC 15886 T collected at logarithmic growth and stationary phases were inoculated into fresh modified MRS medium with 11 U/ml of the bacteriocin, negligible cell growth was observed, irrespective of different growth phases of cells inoculated. Little or no increase in cell concentrations in the media containing bacteriocin showed that the action of the bacteriocin produced by NPSI 38 was bacteriostatic against the hiochi-bacterium.

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  • Production of extracellular bifidogenic growth stimulator (BGS) from Propionibacterium shermanii using a bioreactor system with a microfiltration module and an on-line controller for lactic acid concentration Reviewed

    Tomoaki Kouya, Kazuhiro Tobita, Masahito Horiuchi, Eri Nakayama, Hiroyoshi Deguchi, Takaaki Tanaka, Masayuki Taniguchi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   105 ( 3 )   184 - 191   2008.3

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    Production of a bifidogenic growth stimulator (BGS) by Propionibacterium freudenreichii subsp. shermanii (Propionibaeterium shermanii) using lactic acid as a carbon source was investigated using different cultivation methods. When a continuous bioreactor system with a filtration device was used at a dilution rate of 0.075 h(-1), the average BGS concentration was 2.4 mg/l, which corresponds to a BGS productivity per cultivation time of 1.8x10(-1)mg(.)l(-1.)h(-1). The BGS productivity per cultivation time in continuous cultivation with filtration was 1.9-fold that (9.4x10(-2) mg(.)l(-1.)h(-1)) in a conventional batch cultivation. In fed-batch cultivation with feed-back control using an online lactic acid controller with a lactic acid biosensor, it was possible to prevent substrate inhibition by maintaining the lactic acid concentration in culture broth low at 3.3 g/l, and an enhanced BGS production (31 mg/l) was successfully attained. The BGS productivity per cultivation time (2.1x 10(-1) mg (.)l(-1.)h(-1)) in the fed-batch cultivation with feed-back control was 2.2-fold that in the conventional batch cultivation. A new bioreactor system was developed by coupling a continuous bioreactor system with a filtration device to an on-line lactic acid controller. Using the new bioreactor system, we produced BGS continuously at a high level of 47 mg/l. The BGS productivities per cultivation time (3.5 mg(.)l(-1.)h(-1)) and the total volume of medium used (1.7x 10(-1) mg(.)l(-1.)h(-1)) obtained in the new bioreactor system were 37-fold and 2.1-fold those in the conventional batch cultivation, respectively. These results described above clearly demonstrate the positive effects of both the continuous filtration for removal of metabolites (propionic and acetic acids) inhibitory to cell growth and feed-back control of lactic acid concentration in the culture broth on BGS production by P shermanii. This paper is the first report on BGS production by the propionic acid bacterium using lactic acid as a carbon source.

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  • Production of Bifidogenic Growth Stimulator by Co-cultivation of Propionibacterium shermanii and Lactobacillus Strain Using Lactose as a Carbon Source Reviewed

    Tomoaki Kouya, Masahiro Horiuchi, Kazuhiro Tobita, Katsuhiro Misawa, Eri Nakayama, Hiroyoshi Deguchi, Takaaki Tanaka, Masayuki Taniguchi

    JOURNAL OF CHEMICAL ENGINEERING OF JAPAN   41 ( 6 )   492 - 496   2008

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    Production of a bifidogenic growth stimulator (BGS) by propionibacterial strains was investigated using lactic acid as a carbon source. The maximum concentration (1.9 mg/L) of BGS was obtained in the mono-cultivation of Propionibacterium shermanii using 20 g/L of lactic acid. The co-cultivation, in which lactose was gradually converted only to lactic acid by a homo-fermentative lactic acid bacterium and subsequently the lactic acid was used as a carbon source for BGS production by P. shermanii, resulted in the successful BGS production. The BGS concentration obtained in the co-cultivations P. shermanii and Lactobacillus casei or P. shermanii and Lactobacillus bulgaricus seas 1.8 or 6.4 mg/L, respectively.

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  • Production of extracellular bifidogenic growth stimulator by anaerobic and, aerobic cultivations of several propionibacterial strains Reviewed

    Tomoaki Kouya, Katsuhiro Misawa, Masahito Horiuchi, Eri Nakayama, Hiroyoshi Deguchi, Takaaki Tanaka, Masayuki Taniguchi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   103 ( 5 )   464 - 471   2007.5

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    Production of a bifidogenic growth stimulator (BGS) by propionic acid bacteria was investigated under anaerobic and aerobic culture conditions. To measure the concentration of extracellular BGS produced by propionic acid bacteria, we evaluated the effects of bioassay conditions using Bifidobacterium longum as a test microorganism on the formation of a growth-stimulation zone. The diameter of the growth-stimulation zone was significantly affected by both the component concentrations and the pH of a bioassay medium. The optimum component concentrations and pH of a bioassay medium were one-half of the normal values and 8.5, respectively. Using the bioassay method, we can measure the concentration of BGS produced by propionic acid bacteria ranging in concentrations from 0.1 mu g/l to 1 mg/l using 1,4-dihydroxy-2-naphthoic acid (DHNA) and 2-amino-3-carboxy-1,4-naphthoquinone (ACNQ) as standards. Of six dairy propionic acid bacterial strains tested, the four strains (Propionibacterium freudenreichii ET-3, P. shermanii PZ-3, P. acidipropionici JCM 6432, and P. jensenii JCM 6433) produced BGS at a concentration range of 4-23 mg/l under the anaerobic culture conditions. Analysis of high performance liquid chromatography (HPLC) showed that more than 70% of total BGS produced in supernatant samples was DHNA and no ACNQ was produced by the strains. The effect of oxygen supply on BGS production was investigated for the four BGS-producing strains. The aerobic conditions exerted in positive effects on BGS production by only P. acidipropionici JCM 6432. The concentration of BGS obtained in the aerobic cultivation of P. acidipropionici JCM 6432 was 1.3-fold than that in anaerobic cultivation. Different properties (BGS production as well as cell growth and glucose metabolism) occurring in response to the aerobic conditions were observed, depending on the propionic acid bacterial strain used. This paper is the first report on BGS production by propionibacterial strains except for P. freudenreichii.

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  • Production of laccase by membrane-surface liquid culture of Trametes versicolor using a poly(L-lactic acid) membrane Reviewed

    Takaaki Tanaka, Satoko Eguchi, Takashi Aoki, Toshiyuki Tamura, Hiroshi Saitoh, Masayuki Taniguchi, Hitomi Ohara, Kazuhiro Nakanishi, Douglas R. Lloyd

    BIOCHEMICAL ENGINEERING JOURNAL   33 ( 2 )   188 - 191   2007.2

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    Membrane-surface liquid culture (MSLC) is a promising method for the bioproduction of highly aerobic filamentous fungi [A. Ogawa, A. Yasuhara, T. Tanaka, T. Sakiyama, K. Nakanishi, Production of neutral protease by membrane-surface liquid culture of Aspergillus oryzae IAM2704, J. Ferment. Bioeng. 80 (1995) 35-40]. This paper reports on the production of laccase by Trametes versicolor on a microporous membrane of poly(L-lactic acid) (PLLA), which can be biodegraded via composting after use. The membrane was stable as a support for 24 days at 30 degrees C. During the first 9 days in MSLC, the fungus produced half as much laccase as it did in liquid-surface culture (LSC); however, the mycelium on the membrane was able to be re-used five times for laccase production. The laccase production was accelerated in the repeated use of the culture while the mycelium in LSC ceased to produce the enzyme. This study shows that compostable PLLA microporous membranes can be used for enzyme production by MSLC of filamentous fungi. (c) 2006 Elsevier B.V. All rights reserved.

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  • プロピオン酸菌の嫌気培養と好気培養によるビフィズス菌増殖因子生産の比較解析

    高屋 朋彰, 出口 央視, 田中 孝明, 谷口 正之

    化学工学会 研究発表講演要旨集   2007   811 - 811   2007

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    DOI: 10.11491/scej.2007f.0.811.0

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  • 乳酸菌バクテリオシンによる清酒製造工程における火落菌の殺菌法の開発

    石山 洋平, 高田 剛臣, 小磯 裕介, 金桶 光起, 渡邊 健一, 田中 孝明, 谷口 正之

    化学工学会 研究発表講演要旨集   2007   812 - 813   2007

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    DOI: 10.11491/scej.2007f.0.812.0

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  • ポリブチレンサクシネート濾過膜の開発

    高橋 真記, 川口 滋子, 齋藤 浩, 谷口 正之, Lloyd D. R., 田中 孝明

    化学工学会 研究発表講演要旨集   2007   535 - 535   2007

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  • Microfiltration membrane of polymer blend of poly(L-lactic acid) and poly(epsilon-caprolactone) Reviewed

    T Tanaka, T Tsuchiya, H Takahashi, T Masayuki, DR Lloyd

    DESALINATION   193 ( 1-3 )   367 - 374   2006.5

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    Biodegradable filtration membranes can be applied in food and biochemical processes to clarify products or to yield particles from suspensions, and then degrade in composting processes. In this study we developed microfiltration membranes from a polymer blend of poly(F-caprolactone) (PCL) and poly(L-lactic acid) (PLLA). The membranes were formed via the thermally induced phase separation process and were used to separate yeast cells from their suspension. The blend ratio (PCL:PLLA) of 4:1 was important to prepare the biodegradable polymer-blend membrane with the ability of the retention of yeast cells and without exfoliation of the membrane. The permeation resistance of PCL-PLLA blend (4: 1) membranes was as high as that of PCL membranes and was three-order lower than that of a PLLA membrane. In the filtration of yeast cell suspensions the increase of the permeation resistance during the filtration was much lower with a PCL-PLLA (4: 1) membrane than with a PLLA membrane. The PCL-PLLA (4: 1) membrane captured the cells as a depth filter while the PLLA membrane did as a screen filter.

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  • Formation of biodegradable polyesters membranes via thermally induced phase separation Reviewed

    T Tanaka, T Tsuchiya, H Takahashi, M Taniguchi, H Ohara, DR Lloyd

    JOURNAL OF CHEMICAL ENGINEERING OF JAPAN   39 ( 2 )   144 - 153   2006.2

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    Microfiltration membranes that can be composted after service were developed from biodegradable polyesters, poly(L-lactic acid) (PLLA) and poly(epsilon-caprolactone) (PCL). The membranes were formed via the thermally induced phase separation method. A 10 wt% PLLA solution in a mixed diluent of 1,4-dioxane and water (87:13 by weight) was prepared in a flat mold and quenched from 52 degrees C (4 degrees C above cloud point temperature) to 0 degrees C. After diluent extraction, the membrane separated yeast cells (6 mu m) from their suspensions. A PCL membrane formed by the same method did not reject yeast cells. PCL membranes formed by quenching a 16 wt% PCL solution to 0 degrees C and quenching a 10 wt% PCL solution to -196 degrees C did separate yeast cells from their suspensions. The permeation flux was much higher in the filtration of 1 kg.m(-3) yeast cell suspension with the PLLA and PCL membranes formed by quenching a 10 wt% PLLA or PCL solution to -196 degrees C than in the filtration with the PLLA membrane formed by quenching a 10 wt% PLLA solution to 0 degrees C. The higher flux would be due to the lower resistance of the membranes formed by liquid nitrogen quenching (-196 degrees C) and the mode of depth filtration. Porous biodegradable microfiltration membranes prepared from these polymers have the potential to serve as disposable filters in food and biochemical industries.

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  • Production of D-lactic acid from defatted rice bran by simultaneous saccharification and fermentation Reviewed

    T Tanaka, M Hoshina, S Tanabe, K Sakai, S Ohtsubo, M Taniguchi

    BIORESOURCE TECHNOLOGY   97 ( 2 )   211 - 217   2006.1

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    Production of D-lactic acid from rice bran, one of the most abundant agricultural by-products in Japan, is studied. Lactobacillus delbrueckii subsp. delbrueckii IFO 3202 and defatted rice bran powder after squeezing rice oil were used for the production. Since the rice bran contains polysaccharides as starch and cellulose, we coupled saccharification with amylase and cellulase to lactic acid fermentation. The indigenous bacteria in the rice bran produced racemic lactic acid in the saccharification at pH 6.0-6.8. Thus the pH was controlled at 5.0 to suppress the growth of the indigenous bacteria. L. delbrueekii IFO 3202 produced 28 kg m(-3) lactic acid from 100 kg m(-3) rice bran after 36 h at 37 degrees C. The yield based on the amount of sugars soluble after 36-h hydrolysis of the bran by amylase and cellulase (36 kg m(-3) from 100 kg m(-3) of the bran) was 78%. The optical purity of produced D-lactic acid was 95% e.e. (c) 2005 Elsevier Ltd. All rights reserved.

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  • 麹汁培地を用いて生産した乳酸菌バクテリオシンによる清酒の火落ち防止 Reviewed

    石山 洋平, 中西 利公, 高田 剛臣, 金桶 光起, 渡邊 健一, 成田 護, 田中 孝明, 谷口 正之

    化学工学会 研究発表講演要旨集   2006 ( 0 )   402 - 402   2006

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  • 精米残渣の高速酵素糖化とその発酵原料としての有効利用 Reviewed

    樋口 裕樹, 加藤 壮司, 尾崎 雄一, 金本 繁晴, 田中 孝明, 谷口 正之

    化学工学会 研究発表講演要旨集   2006 ( 0 )   403 - 403   2006

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  • チーズ製造用プロピオン酸菌の好気培養によるBGS生産とその代謝変動の解析

    高屋 朋彰, 中山 惠理, 出口 央視, 田中 孝明, 谷口 正之

    化学工学会 研究発表講演要旨集   2006   401 - 401   2006

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    DOI: 10.11491/scej.2006f.0.401.0

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  • 表面凹凸構造により実効濾過面積を拡大した生分解性プラスチック濾過板の開発

    田中 孝明, 指方 太陽, 青木 卓史, 齋藤 浩, 谷口 正之, Lloyd D. R.

    化学工学会 研究発表講演要旨集   2006   258 - 258   2006

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  • 加水分解酵素-酸化酵素-複合酵素系を用いた女性ホルモン及びその抱合体の酵素処理

    田中 孝明, 石崎 雄一, 田村 俊幸, 川瀬 智一, 谷口 正之

    化学工学会 研究発表講演要旨集   2006   944 - 944   2006

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  • Production of L-lactic acid by simultaneous saccharification and fermentation using unsterilized defatted rice bran as a carbon source and nutrient components Reviewed

    M Taniguchi, M Hoshina, S Tanabe, Y Higuchi, K Sakai, S Ohtsubo, K Hoshino, T Tanaka

    FOOD SCIENCE AND TECHNOLOGY RESEARCH   11 ( 4 )   400 - 406   2005.12

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    On the basis of growth rate at low PH, yield of lactic acid from glucose, and optical purity of lactic acid produced, we selected lactic acid bacteria favorable for production of optically pure L-lactic acid from defatted rice bran without sterilization. Of 21 strains tested, strains Nos.13 and 16 produced 27-29 kg m(-3) of lactic acid with high optical purity from 100 kg m(-3) Of unsterilized defatted rice bran in simultaneous saccharification and fermentation (SSF) with MRS medium at PH 4.5, a level at which the growth of indigenous lactic acid bacteria in defatted rice bran was suppressed. In a SSF process using strain No.16 in which McIlvaine buffer (PH 4.5) was used instead of MRS medium, no growth of indigenous lactic acid bacteria was observed in defatted rice bran, and 28kg m(-3) of lactic acid with 92% L-type content was produced from 100 kg m(-3) of unsterilized defatted rice bran. In SSF using McIlvaine buffer (PH 4.5), the protein fraction of defatted rice bran was found to play a significant role as a nitrogen source for the growth of lactic acid bacteria. By increasing the initial cell concentration to OD660=1.0 for SSF using strain No. 16 and McIlvaine buffer (PH 4.5), the proportion of L-lactic acid produced was enhanced to 95%.

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  • Evaluation of pretreatment with Pleurotus ostreatus for enzymatic hydrolysis of rice straw Reviewed

    M Taniguchi, H Suzuki, D Watanabe, K Sakai, K Hoshino, T Tanaka

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   100 ( 6 )   637 - 643   2005.12

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    The effects of biological pretreatment of rice straw using four white-rot fungi (Phanerochaete chrysosporium, Trametes versicolor, Ceriporiopsis subvermispora, and Pleurotus ostreatus) were evaluated on the basis of quantitative and structural changes in the components of the pretreated rice straw as well as susceptibility to enzymatic hydrolysis. Of these white-rot fungi, P ostreatus selectively degraded the lignin fraction of rice straw rather than the holocellulose component. When rice straw (water content of 60%) was pretreated with P ostreatus for 60 d, the total weight loss and the degree of Klason lignin degraded were 25% and 41%, respectively. After the pretreatment, the residual amounts of cellulose and hemicellulose were 83% and 52% of those in untreated rice straw, respectively. By enzymatic hydrolysis with a commercial cellulase preparation for 48 111, 52% holocellulose and 44% cellulose in the pretreated rice straw were solubilized. The net sugar yields based on the amounts of holocellulose and cellulose of untreated rice straw were 33% for total soluble sugar from holocellulose and 32% for glucose from cellulose. The SEM observations showed that the increase in susceptibility of rice straw to enzymatic hydrolysis by pretreatment with P ostreatus is caused by partial degradation of the lignin seal. When the content of Klason lignin was less than 15% of the total weight of the pretreated straw, enhanced degrees of enzymatic solubilization of holocellulose and cellulose fractions were observed as the content of Klason lignin decreased.

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  • デプスフィルターによる菌体の濾過 Reviewed

    田中 孝明, 三村 朋也, 古賀 光雄, 吉田 政晴, 谷口 正之, 中西 一弘

    化学工学会 研究発表講演要旨集   2005 ( 0 )   408 - 408   2005

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  • 未殺菌米ぬかを原料とした乳酸発酵の低コスト化 Reviewed

    樋口 裕樹, 渡邊 大助, 保科 昌宏, 田中 孝明, 谷口 正之

    化学工学会 研究発表講演要旨集   2005 ( 0 )   721 - 721   2005

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  • 担子菌によって脱リグニン処理した稲わらの乳酸とエタノールへの微生物変換 Reviewed

    渡邊 大助, 鈴木 弘之, 樋口 裕樹, 田中 孝明, 谷口 正之

    化学工学会 研究発表講演要旨集   2005 ( 0 )   720 - 720   2005

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  • Production of L-lactic acid from a mixture of xylose and glucose by co-cultivation of lactic acid bacteria Reviewed

    M Taniguchi, T Tokunaga, K Horiuchi, K Hoshino, K Sakai, T Tanaka

    APPLIED MICROBIOLOGY AND BIOTECHNOLOGY   66 ( 2 )   160 - 165   2004.12

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    The production of optically pure lactic acid in a high yield from xylose or a mixture of xylose and glucose, which is a model hydrolysate of lignocellulose, is described. In a single cultivation, Enterococcus casseliflavus produced 38 g/l of lactic acid with an optical purity of 96% enantiomeric excess (ee) and 6.4 g/l of acetic acid from 50 g/l of xylose when MRS medium was used. When a mixture of 50 g/l of xylose and 100 g/l of glucose was used as the carbon source in a cultivation of E. casseliflavus alone, glucose was converted to lactic acid in the early phase of the cultivation but xylose was hardly consumed. In a co-cultivation where E. casseliflavus and Lactobacillus casei specific for glucose were simultaneously inoculated, little or no lactic acid was produced after the glucose was almost consumed. A co-cultivation with two-stage inoculation ( in which E. casseliflavus was added at a cultivation time of 40 h after L. casei cells were inoculated) resulted in complete consumption of 50 g/l of xylose and 100 g/l of glucose. In the co-cultivation, 95 g/l of lactic acid with a high optical purity of 96% ee was obtained at 192 h. Such a co-cultivation using two microorganisms specific for each sugar is considered to be one promising cultivation technique for the efficient production of lactic acid from a sugar mixture derived from lignocellulose.

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  • Formation of poly(L-lactic acid) microfiltration membranes via thermally induced phase separation Reviewed

    T Tanaka, DR Lloyd

    JOURNAL OF MEMBRANE SCIENCE   238 ( 1-2 )   65 - 73   2004.7

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    Microfiltration membranes of poly(L-lactic acid) (PLLA) were prepared from PLLA-1,4-dioxane-water solutions via the thermally induced phase separation process. Ternary phase diagrams for this system, determined at 10, 48, and 80 C, indicate that essentially polymer-free droplets form in a matrix phase of PLLA-1,4-dioxane-water solution. Rapid solidification of the PLLA after the initial liquid-liquid phase separation was necessary to obtain membranes with open pores at the membrane surface and low permeation resistance. Using filtration of cell suspensions, the effective pore size of the best membrane formed in this study was found to be between 0.6 and 4.4 mum. (C) 2004 Elsevier B.V. All rights reserved.

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  • Clarification of interactions among microorganisms and development of co-culture system for production of useful substances Reviewed

    M Taniguchi, T Tanaka

    RECENT PROGRESS OF BIOCHEMICAL AND BIOMEDICAL ENGINEERING IN JAPAN I   90   35 - 62   2004

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    Co-culture systems containing two microorganisms for the production of useful substances are described. We developed a novel co-culture system composed of two fermentors and two microfiltration modules. The proposed co-culture system allowed regulation of the dissolved oxygen concentration at a level suitable for an individual microorganism in each fermentor, as well as the successful exchange of culture medium between two fermentors. By co-culture, using a combination of Pichia stipitis and Saccharomyces cerevisiae, ethanol was produced efficiently from a mixture of glucose and xylose. Moreover, the useful probiotic cells were simultaneously produced with a high productivity by our co-culture using a combination of Bifidobacterium and Propionibacterium. Kefiran production by Lactobacillus kefiranofaciens alone under the culture conditions, established by mimicking the presence and activities of yeast cells in kefir grains, was also investigated. The results obtained showed that under the culture conditions established by mimicking the actions of yeast cells on L. kefiranofaciens in kefir grains, the amount of kefiran produced was enhanced, even when the lactic acid bacterium alone was used.

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  • Treatment of nonylphenol with laccase in a rotating reactor Reviewed

    T Tanaka, M Nose, A Endo, T Fujii, M Taniguchi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   96 ( 6 )   541 - 546   2003.12

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    The effects of operation conditions on reaction rate in the enzymatic oxidative treatment of nonylphenol with laccase in a rotating reactor were studied. Sea sand that adsorbed nonylphenol was used as a polluted soil model. Nonylphenol concentration decreased with laccase treatment five times faster at 40degreesC than at 10degreesC and the apparent activation energy of the enzyme reaction was 39 kJ mol(-1), which was in the range of the values reported for similar laccase reactions. Reaction rate increased when the angle of the axis of the rotating reactor from the vertical line increased and when the speed of revolution was increased to 10 rpm at different volumes of the enzyme solution. Thus, mixing is important for the oxidation of nonylphenol with laccase. The nonylphenol released from the sand into the enzyme solution in the initial stage of the treatment was easily oxidized. However, the nonylphenol adsorbed on the sand reacted slowly. Reaction rate increased nearly proportional to the square root of enzyme concentration, which suggests that the nonylphenol radical reacted with unoxidized nonylphenol (nonenzymatic propagation) during the enzymatic oxidative treatment. The dependence of reaction rate on the nonylphenol concentration was similar to the Michaelis-Menten type. The residual estrogenic activity of the treated sand was measured by medaka vitellogenin assay. The estrogenic activity decreased to 1/6-1/90 after 24 h of the treatment.

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  • Enzymatic oxidative polymerization of 4-chloroguaiacol by laccase Reviewed

    T Tanaka, M Yamamoto, M Takahashi, T Fujii, M Taniguchi

    JOURNAL OF CHEMICAL ENGINEERING OF JAPAN   36 ( 9 )   1101 - 1106   2003.9

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    The kinetics of enzymatic oxidative polymerization of 4-chloroguaiacol by laccase was studied. 4-Chloroguaiacol, a model compound of the chloroguaiacols formed in the bleaching process in the pulp industry, was oxidized by laccase from Trametes sp. Ha1 to eliminate chloroguaiacols from pulp wastewater by polymerization and precipitation. On the basis of the reaction rate obtained from the decrease in the 4-chloroguaiacol concentration, the optimum pH of the reaction was 4-5 and the apparent Michaelis constant K. for the reaction was more than 5 mmol dm(-3). However, the value of K. obtained from the dependence of oxygen consumption rate on the concentration of 4-chloroguaiacol was 0.092 mmol dm(-3). The reaction rate was expressed as the sum of the enzymatic oxidation rate of 4-chloroguaiacol and the polymerization rate of the radical formed in the enzymatic oxidation. The kinetics of the enzymatic oxidative polymerization shown in this study will be useful to understand those of the enzymatic oxidation of other phenolic compounds.

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  • Relationship between cake structure and membrane pore size in crossflow filtration of microbial cell suspension containing fine particles Reviewed

    T Tanaka, Y Yamagiwa, T Nagano, M Taniguchi, K Nakanishi

    JOURNAL OF CHEMICAL ENGINEERING OF JAPAN   34 ( 12 )   1524 - 1531   2001.12

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    The structural change of a filter cake and its effect on permeation flux in crossflow filtration of a suspension containing particles different in size were studied. A suspension containing yeast cells (5 mum in average diameter) and polystyrene latex particles (0.5 mum) with different compositions was used as a model. The change in the structure of the filter cake formed on the membrane surface was observed by using a scanning electron microscope. When the pore size was smaller than the latex particle size, a yeast cell layer containing latex particles was formed on the membrane at the initial stage of filtration. Then, the yeast cell layer was replaced by the latex particle layer. The increase of the circulation flow rate decreased the amount of the filter cake to increase the permeation flux in this case. On the other hand, when the membrane pore size was larger than the latex particle size, a yeast cell layer was first formed on the membrane surface followed by deposition of latex particles on it. The permeation flux was relatively higher than the former case. However, the permeation flux slightly increased with the increase of the circulation flow rate although the weight of the filter cake decreased in the latter case. Thus, a small amount of the latex particles in the cell suspension considerably decreased the permeation flux in crossflow filtration of the suspension. In addition the dependence of the permeation flux on the circulation flow rate was proved to be different whether the small particles were smaller than the membrane pores. These findings including the structure of the filter cakes will be helpful to understand the permeation behaviors in the crossflow filtration of microbial broths which usually contain small particles.

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  • Treatment of Model Soils Contaminated with Phenolic Endocrine Disrupting Chemicals by Laccase from Trametes sp. in a Rotating Reactor Reviewed

    T. Tanaka, T. Tonosaki, M. Nose, N. Tomidokoro, N. Kadomura, T. Fujii, M. Taniguchi

    Journal of Bioscience and Bioengineering   92 ( 4 )   312 - 316   2001

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  • Kefiran Production by Lactobacillus kefiranofaciens under the Culture Conditions Established by Mimicking the Existence and Activities of Yeast in Kefir Grains Reviewed

    Masayuki Taniguchi, Masashi Nomura, Takahiro Itaya, Takaaki Tanaka

    Food Science and Technology Research   7 ( 4 )   333 - 337   2001

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    Kefiran production by lactobacillus kefiranofaciens alone under the culture conditions established by mimicking the presence and activities of yeast cells in kefir grains was investigated. When the pH of the culture broth was controlled at pH 5.5 by adding a 4 N-NaOH solution during cultivation, cell growth and kefiran production were stimulated as compared with those in the cultivation without pH control. The addition of 5 g/l of yeast extract to the medium was essential for kefiran production. By sparging a mixed gas of N2 and CO2 at a volume ratio of 9:1 into the culture broth at 0.3 vvm throughout the cultivation, a slight increase in the amount (670 mg/l) of kefiran produced was observed as compared with that (650 mg/l) in the cultivation without aeration. When 10 g/l or 20 g/l of ethanol was added to the medium containing 50 g/l of lactose, the kefiran concentrations were about 930 mg/l or 840 mg/l at 8 days, respectively. The maximum concentration of kefiran obtained was about 1040 mg/l at 10 days, when 10 g/l of ethanol was added to the medium containing 75 g/l of lactose. These results showed that under the culture conditions established by mimicking the actions of yeast cells on L. kefiranofaciens in kefir grains the amount of kefiran produced was enhanced even when only the lactic acid bacterium was used.

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  • Enzymatic degradation of alkylphenols, bisphenol A, synthetic estrogen and phthalic ester Reviewed

    T Tanaka, K Yamada, T Tonosaki, T Konishi, H Goto, M Taniguchi

    WATER SCIENCE AND TECHNOLOGY   42 ( 7-8 )   89 - 95   2000

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    Five endocrine disrupting chemicals were degraded enzymatically in model bottom sediments. Since alkylphenols, bisphenol A, synthetic estrogen and phthalic esters are hydrophobic and found to be in bottom sediments in the water environment, we used a modal polluted sediment in which those chemicals were adsorbed on sea sand. The concentration of alkylphenols (nonylphenol and octylphenol) and synthetic estrogen (ethynylestradiol) were decreased by fungal laccases which oxidize phenols with the aid of molecular oxygen. Bisphenol A was degraded by laccase from basidiomycetes with the aid of extracellular polymers. Dibutyl phthalate was hydrolyzed by lipase from Candida cylindracea. These results will be useful in the remediation of the bottom sediments polluted by endocrine disrupting chemicals.

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  • Production of a mixture of antimicrobial organic acids from lactose by co-culture of Bifidobacterium longum and Propionibacterium freudenreichii Reviewed

    M Taniguchi, H Nakazawa, O Takeda, T Kaneko, K Hoshino, T Tanaka

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   62 ( 8 )   1522 - 1527   1998.8

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    The antimicrobial activities of standard solutions of three organic acids (lactic, acetic, and propionic acids) were compared using Micrococcus luteus, Pseudomonas sp. and Staphylococcus aureus as test microorganisms. At the same concentrations of the undissociated form, the antimicrobial activities of acetic and propionic acids were higher than that of lactic acid, irrespective of test microorganisms. In a single cultivation of Bifidobacterium longum, a mixture of lactic (17 g/l) and acetic (20 g/l) acids was produced from 50 g/l lactose and its antimicrobial activities against M. luteus, Pseudomonas sp., and S. aureus correspond to that of 32, 19, and 25 g/l of acetic acid, respectively. To increase the total antimicrobial activity, a co-culture of B. longum and Propionibacterium freudenreichii, in which lactic acid produced once from lactose by B. longum was converted to acetic and propionic acids by P. freudenreichii, was done using TPY medium containing commercially available peptones as a nitrogen source. By the sequential conversion of lactose using the two microorganisms, the culture supernatant containing a mixture of acetic (27 g/l) and propionic (13 g/l) acids without lactic acid was produced. The antimicrobial activities of the mixture against nl. luteus, Pseudomonas sp., and S. aureus were 35, 30, and 26 g/l as a concentration of acetic acid, respectively, higher than that obtained in the cultivation of B. longum alone. When the medium containing an enzymatic hydrolyzate of whey proteins with a protease was used in the co-culture of B. longum and P. freudenreichii, the culture supernatant containing the mixture of organic acids was also obtained in the same manner as the co-culture using TPY medium and the activities were 43, 29, and 29 g/l as a concentration of acetic acid for M. luteus, Pseudomonas sp. and S. aureus, respectively.

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  • Formation of the gel layer of polymers and its effect on the permeation flux in crossflow filtration of Corynebacterium glutamicum broth Reviewed

    T Tanaka, K Usui, K Nakanishi

    SEPARATION SCIENCE AND TECHNOLOGY   33 ( 5 )   707 - 722   1998

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    The behavior of the permeation flux in crossflow filtration of Corynebacterium glutamicum broth was studied. In the beginning of filtration the permeation flux changed in accordance with the cake filtration model. The permeation flux agreed with the value calculated from the weight of the cell layer formed on the membrane per unit filtration area and the specific resistance of the cell layer measured in dead-end filtration. After cell deposition on the membrane, permeation resistance rapidly increased. The increase was due to the formation of the gel layer of the proteins and polysaccharides in the supernatant of the broth. When the circulation flow rate was raised, the weight of the cell layer reduced. However, the beginning of the rapid increase of permeation resistance came earlier and the apparent specific resistance of the cell layer increased. The higher the circulation flow rate, the higher the permeation flux at the beginning of the rapid increase of resistance. When the transmembrane pressure was lowered or the cell concentration was raised, the beginning of the rapid increase of the permeation resistance came earlier. The permeation flux at which the formation of the gel layer began was almost constant, even if the transmembrane pressure and the cell concentration were changed.

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  • Characteristics in crossflow filtration using different yeast suspensions Reviewed

    T Tanaka, S Tsuneyoshi, W Kitazawa, K Nakanishi

    SEPARATION SCIENCE AND TECHNOLOGY   32 ( 11 )   1885 - 1898   1997

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    Characteristics of crossflow filtration of cell types were compared. Although the cell sizes were similar for the six yeast cells, the specific resistance of the cake measured in dead-end filtration and permeation flux were different. The flux decreased with increasing specific resistance. However, the dependencies of the steady-state flux on the shear stress were similar for all the yeast cells. The steady state permeation flux of the yeast cell suspensions was correlated with the shear stress to the 0.6-0.9th power and with the cell concentration to the -0.3rd power regardless of the transmembrane pressure. The permeation flux in the unsteady state was well simulated by the correlation for the lift velocity of the cells obtained from the steady-state flux.

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  • Filtration behaviors of rod-shaped bacterial broths in unsteady-state phase of cross-flow filtration Reviewed

    T Tanaka, K Usui, K Kouda, K Nakanishi

    JOURNAL OF CHEMICAL ENGINEERING OF JAPAN   29 ( 6 )   973 - 981   1996.12

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    Filtration behaviors in the unsteady-state phase of crossflow filtration of broths of Bacillus subtilis, Escherichia coli, and Lactobacillus delbrueckii, which are rod-shaped, were studied from the viewpoint of the changes in the specific resistance and in the structure of the microbial cake formed on the membrane surface. The permeation flux followed the cake filtration law at the initial stage of the crossflow filtration of the broths of B. subtilis and E. coli, where the cells deposited randomly on the membrane. filtration of at. delbrueckii broth, the period of random deposition was shorter. cake formed at the initial stage agreed with that measured in dead-end filtration. started to increase in comparison with that measured in dead-end filtration due to shear-induced arrangement of the cells. The extent of the increase in specific resistance became higher and the time taken to start the cell arrangement became shorter with increasing circulation flow rate. The increase in specific resistance due to the shear-induced arrangement was more appreciable in the crossflow filtration of the broth oft. delbrueckii than that of B. subtilis and E. coli. The average permeation flux was increased considerably by applying periodical backwashing with appropriate time intervals. The permeation flux was well predicted by the cake filtration law, since the cells deposited in a way similar to that for dead-end filtration during a sufficiently short period of crossflow filtration in a backwashing mode.

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  • CROSS-FLOW FILTRATION OF BAKERS-YEAST WITH PERIODICAL STOPPING OF PERMEATION FLOW AND BUBBLING Reviewed

    T TANAKA, H ITOH, K ITOH, K NAKANISHI, T KUME, R MATSUNO

    BIOTECHNOLOGY AND BIOENGINEERING   47 ( 3 )   401 - 404   1995.8

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    The periodical stopping of permeation flow was applied to increase the permeation flux in crossflow filtration of commercially available baker's yeast cell suspension. The permeation flux after 3 h filtration in the crossflow filtration increased to 8 x 10(-5) m(3)/m(2) s (290 L/m(2) h) from 2 x 10(-5) m(3)/m(2) s (72 L/m(2) h) by applying the periodical stopping of permeation. Introduction of air bubbles during the stopping period of permeation further increased the flux. (C) 1995 John Wiley and Sons, Inc.

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  • SYNTHESIS OF ASPARTAME PRECURSOR WITH AN IMMOBILIZED THERMOLYSIN IN MIXED ORGANIC-SOLVENTS Reviewed

    M MIYANAGA, T TANAKA, T SAKIYAMA, K NAKANISHI

    BIOTECHNOLOGY AND BIOENGINEERING   46 ( 6 )   631 - 635   1995.6

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    N-(benzyloxycarbonyl)-L-aspartyl-L-phenylalanine methyl ester, a precursor of the synthetic sweetener, aspartame, was synthesized from N-(benzyloxycarbonyl])-L-aspartic acid and L-phenylalanine methyl ester with an immobilized thermolysin (EC 3.4.24.4) in the mixed organic solvent system of tert-amyl alcohol and ethyl acetate. A mixed solvent consisting of tert-amyl alcohol and ethyl acetate at a ratio of 33:67 (v/v) was found to be the most suitable with respect to synthetic rate and stability of the immobilized enzyme. The reaction continued to proceed quite successfully in a column reactor at 40 degrees C and at a space velocity of 3.6 h(-1) with a yield of 99%, using 40 mM Z-Asp and 200 mM PheOMe dissolved in the mixed solvent as the substrate. (C) 1995 John Wiley & Sons, Inc.

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  • PRODUCTION OF KOJIC ACID FROM ASPERGILLUS-ORYZAE VAR ORYZAE BY MEMBRANE-SURFACE LIQUID CULTURE Reviewed

    A OGAWA, Y MORITA, T TANAKA, T SAKIYAMA, K NAKANISHI

    BIOTECHNOLOGY TECHNIQUES   9 ( 2 )   153 - 156   1995.2

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    Membrane-surface liquid culture (MSLC) developed by the authors (Yasuhara er (al., 1994) was applied to the production of kojic acid, using Aspergillus oryzae var. oryzae IFO 30113. By the fed-batch MSLC with intermittent glucose addition, the amount of kojic acid increased to over 50 times that obtained by means of the culture in shake flasks.

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  • PRODUCTION OF KOJIC ACID BY MEMBRANE-SURFACE LIQUID CULTURE OF ASPERGILLUS-ORYZAE NRRL484 Reviewed

    A OGAWA, Y WAKISAKA, T TANAKA, T SAKIYAMA, K NAKANISHI

    JOURNAL OF FERMENTATION AND BIOENGINEERING   80 ( 1 )   41 - 45   1995

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    Membrane-surface liquid culture (MSLC) was applied to the production of kojic acid using Aspergillus oryzae NRRL484. The characteristics of kojic acid fermentation by MSLC were compared particularly with those by shaking culture. The maximum concentration of kojic acid produced and the production rate of kojic acid by the MSLC were usually higher than those by the shaking culture. In the shaking culture, the kojic acid production was the highest with 0.05-0.25% yeast extract and the maximum concentration was around 20 mg/ml. In the MSLC, the highest kojic acid concentration of about 30 mg/ml was obtained with 0.25-0.5 % yeast extract. By addition of powdered glucose at a final concentration of 10%, 2-3 times at appropriate intervals during the batch MSLC, the concentration of kojic acid increased to over 100 mg/ml and kojic acid crystals formed in the medium. Repeated fed-batch production of kojic acid by MSLC was quite successful. The concentration of kojic acid produced in each batch was maintained at 75 mg/ml or more with a yield of around 50% for 10 batches and 75 d when the medium contained 0.25% yeast extract. The kojic acid productivity by the repeated fed-batch MSLC was 14.2 g/l/d, about 6 times higher than that by the shaking culture with the medium containing 0.25% yeast extract. After the 11th batch, the production rate decreased, probably due to an increased amount of cells formed on the membrane.

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  • PRODUCTION OF NEUTRAL PROTEASE BY MEMBRANE-SURFACE LIQUID CULTURE OF ASPERGILLUS-ORYZAE IAM2704 Reviewed

    A OGAWA, A YASUHARA, T TANAKA, T SAKIYAMA, K NAKANISHI

    JOURNAL OF FERMENTATION AND BIOENGINEERING   80 ( 1 )   35 - 40   1995

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    A membrane-surface liquid culture (MSLC), in which microorganisms were grown on a microporous membrane surface exposed to the air with the other side of the membrane in contact with liquid medium, was applied to the production of neutral protease from Aspergillus oryzae LAM2704. The amount of protease produced by batch MSLC was much higher than that produced by shaking culture, liquid-surface culture, or agar plate culture. In MSLC the amount of protease produced per milligram of dry cells as well as the amount of protease produced were dependent particularly on the glucose concentration. By decreasing the glucose concentration from 1.5% to 0.2%, the amount of protease produced and its specific amount were increased by 60% and around 7 times, respectively. Using a medium containing 0.2% glucose and 0.4% casein, long-term repeated-batch production of the enzyme was conducted by exchanging the medium every 12 to 48 h with a fresh one. Protease production continued in more than 15 successive batches, although the amount of enzyme produced in each batch gradually decreased. When the medium was exchanged every 24 h, the degree of the decrease was the smallest, and the cumulative amount of enzyme produced was more than 20 and 400 times those obtained in the batch MSLC and shaking culture, respectively.

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  • SYNTHESIS OF ASPARTAME PRECURSOR WITH AN IMMOBILIZED THERMOLYSIN IN TERT-AMYL ALCOHOL Reviewed

    T NAGAYASU, M MIYANAGA, T TANAKA, T SAKIYAMA, K NAKANISHI

    BIOTECHNOLOGY AND BIOENGINEERING   43 ( 11 )   1118 - 1123   1994.5

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    N-(Benzyloxycarbonyl)-L-aspartyl-L-phenylalanine methyl ester (Z-AspPheOMe), a precursor of the synthetic sweetner asparatame, was synthesized from N-(benzyloxycarbonyl)-L-aspartic acid (Z-Asp) and L-phenylalanine methyl ester (PheOMe) with an immobilized thermolysin in various organic solvents. We found that in tert-amyl alcohol containing a small amount of water the immobilized enzyme showed a high activity comparable to that in ethyl acetate with quite a high stability. The immobilized enzyme was fully stable up to 70 degrees C in tert-amyl alcohol in the absence of the substrate, and up to 50 degrees C in the presence of the substrate. The high stability in the presence of the substrate was found due to the fact that the release of calcium ions, the stabilizing factor of thermolysin, is suppressed.
    The substrate concentration dependence of th initial synthetic rate with the immobilized enzyme was quite different from that with the free enzyme in the biphasic system, in contrast to that in ethyl acetate. Finally, Z-AspPheOMe was continuously synthesized in a column reactor using 200 mM PheOMe and 120 mM Z-Asp as the substrate for over 300 h at 45 degrees C and a space velocity of 1 h(-1) without any loss of activity. (C) 1994 John Wiley & Sons, Inc.

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  • CROSS-FLOW FILTRATION OF YEAST BROTH CULTIVATED IN MOLASSES Reviewed

    T TANAKA, R KAMIMURA, R FUJIWARA, K NAKANISHI

    BIOTECHNOLOGY AND BIOENGINEERING   43 ( 11 )   1094 - 1101   1994.5

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    A broth of yeast cells cultivated in molasses was cross-filtered with a thin-channel module. The permeation flux gradually decreased at a constant cell concentration. The flux was much lower than that obtained for yeast broth cultivated in yeast extract, polypeptone, and dextrose (YPD) medium during the filtration. The flux did not depend on the membrane pore size (0.45 to 5 mu m). The steady-state flux was one-twentieth that calculated for a cake filtration model from the amount of cake per unit filtration area and the specific resistance of the cake measured in a dead-end filtration apparatus. The lower flux was due to small particles (most of which were less than 1 mu m in diameter) in the molasses. The mechanism of crossflow filtration of broths of yeast cells cultivated in molasses was clarified by analysis of the change in flux with time and observations with scanning electron microscopy. At the initial stage of crossflow filtration the yeast cells and particles from the molasses were deposited on the membrane to form a cake in a similar way to dead-end filtration. After the deposition of cells onto the membrane ceased, the fine particles from molasses formed a thin layer, which had higher resistance than the cake formed next to the membrane. The backwashing method was effective to increase the flux. The flux increased low when the pore size was 0.45 to 0.8 mu m, but using larger pores of 3 to 5 mu m it returned almost to the base line. (C) 1994 John Wiley & Sons, Inc.

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  • SYNTHESIS OF DIPEPTIDE PRECURSORS WITH AN IMMOBILIZED THERMOLYSIN IN ETHYL-ACETATE Reviewed

    T NAGAYASU, M MIYANAGA, T TANAKA, T SAKIYAMA, K NAKANISHI

    BIOTECHNOLOGY AND BIOENGINEERING   43 ( 11 )   1108 - 1117   1994.5

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    N-(Benzyloxycarbonyl)-L-aspartyl-L-phenylalamine methyl ester (Z-AspPheOMe), a precursor of the aspartame, and N-(benzyloxycarbonyl)-L-phenylalanyl-L-phenylalanine methyl ester (Z-PhePheOMe) were synthesized from the respective amino acid derivatives with an immobilized thermolysin (EC 3.4.24.4) in ethyl acetate. Various factors affecting the synthesis of these dipeptide precursors were clarified. The initial synthetic rate was the highest at the water content of 3.5% for both reactions. The substrate concentration dependencies of the initial synthetic rate of Z-AspPheOMe and Z-PhePheOMe with the immobilized enzyme in ethyl acetate were different from those in an aqueous buffer solution saturated with ethyl acetate but similar to those in the aqueous/organic biphasic system using the free enzyme. Particularly, the initial synthetic rate of Z-AspPheOMe increased in order higher than first order with respect to the concentration of L-phenylalanine methyl ester (PheOMe), whereas it decreased sharply with the concentration of N-(benzyloxycarbonyl)-L-aspartic acid (Z-Asp). Such kinetic behavior could be explained by regarding the inside of the immobilized enzyme as being a biphasic mode composed from the organic phase and aqueous phase where the enzymatic reaction takes place. The reaction in the aqueous/organic biphasic system using the free enzyme could be simulated by taking into consideration the partition of the substrate and the initial rate of synthesis in the aqueous buffer saturated with ethyl acetate. Based on this analysis, the rate of reaction with the immobilized enzyme in ethyl acetate could also be predicted. Z-AspPheOMe and Z-PhePheOMe were synthesized by the fed-batch method where the acid component of the substrate was intermittently added during the course of reaction and by the batch method. In the synthesis of Z-AspPheOMe, the synthetic rate and maximum yield of reaction as well as the stability of the immobilized enzyme were higher in the fed-batch reaction than those in the batch reaction. In the synthesis of Z-PhePheOMe, the results obtained by both methods were similar. (C) 1994 John Wiley & Sons, Inc.

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  • PRODUCTION OF NEUTRAL PROTEASE FROM ASPERGILLUS-ORYZAE BY A NOVEL CULTIVATION METHOD ON A MICROPOROUS MEMBRANE

    A YASUHARA, A OGAWA, T TANAKA, T SAKIYAMA, K NAKANISHI

    BIOTECHNOLOGY TECHNIQUES   8 ( 4 )   249 - 254   1994.4

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    A novel cultivation method using a microporous membrane (membrane-surface liquid culture) was developed, in which moulds are grown on the membrane surface with its opposite side being in contact with a liquid medium. The amount of neutral protease from Aspergillus oryzae produced was more than 10 times higher than that produced by the conventional liquid culture.

    DOI: 10.1007/BF00155416

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  • SHEAR-INDUCED ARRANGEMENT OF CELLS IN CAKE DURING CROSS-FLOW FILTRATION OF ESCHERICHIA-COLI-CELLS Reviewed

    T TANAKA, K ABE, K NAKANISHI

    BIOTECHNOLOGY TECHNIQUES   8 ( 1 )   57 - 60   1994.1

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    The specific resistance of the cake formed in the crossflow filtration of Escherichia coli was higher than that formed in the dead-end filtration. The scanning electronmicrographs revealed that the cells in the cake formed in the crossflow filtration were oriented in the direction of the circulation flow, while the cells deposited at random in the dead-end filtration. The shear-induced arrangement of cells might increase the specific resistance of the cake in crossflow filtration.

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  • FILTRATION CHARACTERISTICS AND STRUCTURE OF CAKE IN CROSS-FLOW FILTRATION OF BACTERIAL SUSPENSION

    T TANAKA, K ABE, H ASAKAWA, H YOSHIDA, K NAKANISHI

    JOURNAL OF FERMENTATION AND BIOENGINEERING   78 ( 6 )   455 - 461   1994

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    A suspension of various microorganisms was cross-filtered and the filtration characteristics were studied. In crossflow filtration of Corynebacterium glutamicum, an ellipsoidal-shaped bacterium, the experimental permeation flux was in agreement with the calculated value based on the filtration theory, using the specific resistance measured in dead-end filtration and the amount of cake per unit filtration area. The cells deposited on the membrane in the same manner as in dead;end filtration. On the other hand, in crossflow filtration of Bacillus species, all of which are rod-shaped cells, the cells in the cake formed on the membrane were oriented toward the direction of the circulation flow. This arrangement of the cells increased the specific resistance of the cake, which made the flux lower than the calculated value, using the specific resistance measured in dead-end filtration. Furthermore, we clarified that the degree of the cell arrangement was dependent on the operational conditions in the crossflow filtration of rod-shaped cells.

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  • Factors affecting the performance of crossflow filtration of yeast cell suspension Reviewed

    T Tanaka, K Nakanishi

    DEVELOPMENTS IN FOOD ENGINEERING, PTS 1 AND 2   653 - 655   1994

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  • IMPROVEMENT OF PERFORMANCE FOR CROSS-FLOW MEMBRANE FILTRATION OF PULLULAN BROTH Reviewed

    H YAMASAKI, MS LEE, T TANAKA, K NAKANISHI

    APPLIED MICROBIOLOGY AND BIOTECHNOLOGY   39 ( 1 )   21 - 25   1993.4

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    To improve the performance of cross-flow membrane filtration of pullulan broth from Aureobasidium pullulans, the effect of the cultivation conditions was examined. In particular, the sucrose concentration in the medium was changed over a wide range. By decreasing the sucrose concentration the distribution of morphology of the microbial cells in the broth changed; the yeast-like form became predominant and, as a result, the specific resistance of the microbial cake was lowered. When the broth was fermented with a sucrose concentration of 2.5% or lower, the filtration characteristics were greatly improved by periodic closure of permeation during cross-flow filtration.

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  • CHARACTERISTICS OF CROSS-FLOW FILTRATION OF PULLULAN BROTH Reviewed

    H YAMASAKI, MS LEE, T TANAKA, K NAKANISHI

    APPLIED MICROBIOLOGY AND BIOTECHNOLOGY   39 ( 1 )   26 - 30   1993.4

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    Cross-flow filtration of culture broth from Aureobasidium pullulans, which elaborates pullulan, was done with a thin channel-type module and microfiltration membranes made of different materials and with different pore sizes. Various factors affecting the results of the filtration were studied. The specific resistance of the microbial cake was found to be higher than that of bakers' yeast, the cells of which are about the same size as an A. pullulans cell, and resistance increased with cultivation time. The flux and transmission of pullulan through the membrane decreased with cultivation time as the specific resistance increased. The flux and transmission of pullulan depended on the structure and pore size of the membrane and also on the pH of the broth. With a polysulphone membrane with a nominal pore size of 2.0 mum, transmission was nearly 100% with negligible leakage of cells and the flux was high when the PH of the broth was adjusted to 2.0.

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  • FACTORS AFFECTING THE PERFORMANCE OF CROSS-FLOW FILTRATION OF YEAST-CELL SUSPENSION Reviewed

    T TANAKA, R KAMIMURA, K ITOH, K NAKANISHI, R MATSUNO

    BIOTECHNOLOGY AND BIOENGINEERING   41 ( 6 )   617 - 624   1993.3

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    Factors affecting the performance of crossflow filtration were investigated with a thin-channel module and yeast cells. In crossflow filtration of Saccharomyces cerevisiae cells cultivated with YPD medium (yeast extract, polypeptone, and dextrose) and suspended in saline, a steady state was attained within several minutes when the cell concentration was low and the circulation flow rate was high. The steady-state flux and the change in flux during the initial unsteady state were explained well by conventional filtration theory, with the amount of cake deposited and the mean specific resistance to the cake measured in a dead-end filtration apparatus used in calculation. When the circulation flow rate was lower than a critical value, a part of the channel of the cross-flow filtration module was plugged with cell cake, and thus the steady-state flux was low. In crossflow filtration of suspensions of commercially available baker's yeast, the flux gradually decreased, and the flux after 8 h of filtration was lower than the value calculated by filtration theory. Fine particles contaminating the baker's yeast was responsible for the decrease. A similar phenomenon was observed in crossflow filtration of a broth of S. cerevisiae cells cultivated in molasses medium, which also contains fine particles. The YPD medium, which does not contain such particles, had no effect of the permeation flux during crossflow filtration.

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  • SOME FACTORS AFFECTING THE FORMATION OF GAMMA-CYCLODEXTRIN USING CYCLODEXTRIN GLYCOSYLTRANSFERASE FROM BACILLUS SP AL-6 Reviewed

    K TOMITA, T TANAKA, Y FUJITA, K NAKANISHI

    JOURNAL OF FERMENTATION AND BIOENGINEERING   70 ( 3 )   190 - 192   1990

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    DOI: 10.1016/0922-338X(90)90185-Y

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  • Enzymatic synthesis of L-glutamic acid dimer precursor with immobilized papain in an organic solvent Reviewed

    Takaaki Tanaka, Kenji Tomita, Kazuhiro Nakanishi

    Chemistry Express   5 ( 7 )   525 - 528   1990

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  • CONTINUOUS PRODUCTION OF L-SERINE BY IMMOBILIZED GROWING CORYNEBACTERIUM-GLYCINOPHILUM CELLS Reviewed

    T TANAKA, K YAMAMOTO, S TOWPRAYOON, H NAKAJIMA, K SONOMOTO, K YOKOZEKI, K KUBOTA, A TANAKA

    APPLIED MICROBIOLOGY AND BIOTECHNOLOGY   30 ( 6 )   564 - 568   1989.6

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    DOI: 10.1007/BF00255360

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Books

  • Production, Forming/Molding, and High-Performance Technologies of Poly(lactic acid)

    Hiroshi Uyama, Yu-I Hsu, Takaaki Tanaka, And Others( Role: Joint author)

    CMC Publishing Co.,Ltd.  2024.1  ( ISBN:9784781317649

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  • 食品製造に役立つ食品工学事典

    日本食品工学会編, 田中孝明他( Role: Joint author)

    2020.12 

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  • カーボンナノチューブの表面処理・分散技術と複合化事例

    技術情報協会編, 田中孝明( Role: Joint author)

    技術情報協会  2019.12  ( ISBN:9784861047725

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  • 生分解性プラスチックの素材・技術開発

    望月政嗣, 監修, 田中孝明, 他, 著( Role: Joint author)

    エヌ・ティー・エス  2019.12  ( ISBN:9784860436230

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  • 多孔質フィルム/膜の製造技術

    田中孝明, 佐光貞樹, 富康博, 川島敏行, 石塚紀生, 瀧健太郎, 串﨑義幸, 平井悠司, 下村政嗣, 横山英明, 早川晃鏡, 武野明義, 高橋紳矢, 松田裕行, 小野寺恒信, 笠井均, 及川英俊, 山田一博, 河野公一, 西川聡( Role: Joint author)

    S&T出版  2016.6  ( ISBN:4907002564

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  • 濾過スケールアップの正しい進め方と成功事例集

    技術情報協会編, 田中孝明

    技術情報協会  2014.8  ( ISBN:9784861045370

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  • 化学便覧 応用化学編 第7版

    日本化学会編, 田中孝明

    丸善出版  2014.1  ( ISBN:9784621087596

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  • 濾過プロセスの最適選定と効率改善

    情報機構編, 田中孝明

    情報機構  2010.10  ( ISBN:9784904080610

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  • カーボンナノチューブの精製・前処理と分散・可溶化技術

    技術情報協会編, 田中孝明

    技術情報協会  2009.4  ( ISBN:9784861042829

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  • 産業酵素の応用技術と最新動向

    井上國世監修, 田中孝明

    シーエムシー出版  2009.3  ( ISBN:9784781301082

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  • バイオプロダクション

    化学工学会バイオ部会編, 田中孝明

    コロナ社  2006.5  ( ISBN:9784339067361

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  • 食品工学ハンドブック

    日本食品工学会編, 田中孝明( Role: Joint author)

    朝倉書店  2006.1  ( ISBN:9784254430912

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  • 化学工学の進歩39,粒子・流体系フロンティア分離技術

    化学工学会編, 田中孝明

    槇書店  2005.10  ( ISBN:9784837506881

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  • 生物工学ハンドブック

    日本生物工学会編, 田中孝明

    コロナ社  2005.6  ( ISBN:9784339067347

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  • Clarification of Interactions among Microorganisms and Development of Co-culture System for Production of Useful Substances

    Adv. Biochem. Engin./Biotechnol., Springer  2004 

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  • Treatment of phenolic endocrine-disrupting chemicals by lignin-degrading enzymes.

    Wastewater Treatment Using Enzymes, Research Signpost, Kerala, India  2003 

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  • Permation behaviors in crossflow microfiltration of microbial suspensions containing fine particles and polymers.

    Proceedings of The First Conference of Aseanian Membrane Society, Tokyo, Japan  2002 

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  • Crossflow filtration of lactic acid bacteria: effects of cell shapes and dextran on permeation flux.

    Proceedings of The First Conference of Aseanian Membrane Society, Tokyo, Japan  2002 

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  • Formation and Structural Change of Cake during Crossflow Microfiltration of Microbial Cell Suspension Containing Fine Particles(jointly worked)

    Bioseparation Engineering  2000 

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  • On the stability of immobilized thermolysin in an organic solvent(jointly worked)

    Chemical Engineering Symposium Series 57, The Society of Chemical Engineers, Japan  1997 

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  • Development of membrane-surface liquid culture of molds(jointly worked)

    1995 

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  • Optimization for synthesis of aspartame precursor using an immobilized thermolysin in an organic solvent(jointly worked)

    Chemical Engineering Symposium Series 44, The Society of Chemical Engineers, Japan  1995 

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  • Crossflow filtration of microbial suspensions(jointly worked)

    Continental Press Singapore, Better living through innovative biochemical engineering  1994 

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  • 生物工学実験書

    日本生物工学会編, 田中孝明

    培風館  1992.10  ( ISBN:4563077178

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  • Crossflow membrane filtration of highly viscous microbial broth(jointly worked)

    S. Furusaki, I. Endo, R. Matsuno, Springer Verlag Tokyo, Biochemical Engineering for 2001  1992 

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Awards

  • Outstanding Paper Award, Membrane

    2021.6   Preparation of polyhydroxyalkanoate microfiltration membranes via nonsolvent–induced phase separation methods

    Kazuyoshi Tabata, Toshiki Shibuya, Kousuke Karakida, Akihito Ochiai, Masayuki Taniguchi, Takaaki Tanaka

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    Award type:Honored in official journal of a scientific society, scientific journal 

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  • The 25th Paper Award of The Society for Biotechnology, Japan

    2017.9   The Society for Biotechnology, Japan   AmyI-1–18, a cationic α-helical antimicrobial octadecapeptide derived from α-amylase in rice, inhibits the translation and folding processes in a protein synthesis system

    TANAKA Takaaki

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  • Outstanding Paper Award of MEMBRANE

    2017.5   The Membrane Society of Japan   Preparation of a poly(L-lactic acid) membrane scaffold with open finger-like pores prepared by a nonsolvent-induced phase separation method with the aid of a surfactant

    H. Minbu, T. Kawase, A. Ochiai, M. Taniguchi, T. Tanaka

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  • Award for Excellence to Authors Publishing in Bioscience, Biotechnology, and Biochemisry in 2014

    2015.3   The Japan Society for Bioscience, Biotechnology, and Agrochemistry   Crystal structure of α-amylase from Oryza sativa: molecular insights into enzyme activity and thermostability

    Akihito Ochiai, Hiroshi Sugai, Kazuki Harada, Seiya Tanaka, Yohei Ishiyama, Kosuke Ito, Takaaki Tanaka, Toshio Uchiumi, Masayuki Taniguchi, Toshiaki Mitsui

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  • Paper Award

    2010.8   Japan Society for Food Engineering   Growth-inhibition of Lactobacillus hilgardii, a bacterium related to hiochi, in the mizu-koji process by bacteriocins from lactic acid bacteria

    Y. Ishiyama, T. Takata, T. Nakanishi, M. Kaneoke, K. Watanabe, F. Yanagida, Y. Chen, T. Kouya, T. Tanaka, M. Taniguchi

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  • The 17th Paper Award of The Society for Biotechnology, Japan

    2009.9   The Society for Biotechnology, Japan   Production of extracellular bifidogenic growth stimulator (BGS) from Propionibacterium shermanii using a bioreactor system with a microfiltration module and an on-line controller for lactic acid concentration

    T. Kouya, K. Tobita, M. Horiuchi, E. Nakayama, H. Deguchi, T. Tanaka, M. Taniguchi

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  • Encouragement Award of Japan Society for Food Engineering

    2007.8   Japan Society for Food Engineering   Studies on Membrane Filtration of Food Microbial Cell Suspensions

    Takaaki Tanaka

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Research Projects

  • アスリートの選手生命を救うPRP治療の確立に向けた基盤的研究

    Grant number:22K11496

    2022.4 - 2025.3

    System name:科学研究費助成事業

    Research category:基盤研究(C)

    Awarding organization:日本学術振興会

    牛木 隆志, 望月 友晴, 川瀬 知之, 田中 孝明

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    Grant amount:\4160000 ( Direct Cost: \3200000 、 Indirect Cost:\960000 )

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  • Development of mixed-mode monolith-type adsorption materials and their application to purification process of proteins and peptides

    Grant number:17K06921

    2017.4 - 2020.3

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for Scientific Research (C)

    Awarding organization:Japan Society for the Promotion of Science

    Tanaka Takaaki

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    Grant amount:\4810000 ( Direct Cost: \3700000 、 Indirect Cost:\1110000 )

    In this study mixed-mode monolith-type adsorption materials were developed to be used in separation and purification of proteins and peptides in biopharmaceuticals production. Porous monoliths were prepared from polymers (poly(methyl methacrylate) and poly(lactic acid)) via phase separation methods. Hydroxyapatite particles that have mixed-mode (multimodal) type adsorption property were mainly composited with the monoliths. The prepared monoliths have adsorption abilities toward model proteins. For example, bovine serum albumin and bovine gamma-globulin were separated by one of the developed monoliths.

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  • Developments of scaffolds and processing technology to maximize the osteogenic potential of cultured periosteal sheets

    Grant number:24390443

    2012.4 - 2016.3

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for Scientific Research (B)

    Awarding organization:Japan Society for the Promotion of Science

    Kawase Tomoyuki, OKUDA KAZUHIRO, NAGATA MASAKI, TANAKA TAKAAKI

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    Grant amount:\18460000 ( Direct Cost: \14200000 、 Indirect Cost:\4260000 )

    The purpose of this project is to demonstrate ideal structure and stiffness of scaffolds for osteogenic cells and to develop ideal scaffolds with biodegradable polymer materials. Under regular culture conditions, human periosteal cells (PC) expressed integrin α1β1 and CD44 as major adhesion molecules. Atomospheric plasma treatment modified the titanium surface and facilitated their osteoblastic differentiation. We also found that human platelet-rich fibrin (PRF) extract can be substituted for FBS. As for scaffolding materials, we developed a combinational porous membrane made of polycaprolactone and hydroxyapatite and found its stiffness and microstructure appropriate for PCs.

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  • Control of the internal and surface structures of biodegradable porous membranes with the aid of surfactants and their application to bioprocess

    Grant number:24560957

    2012.4 - 2016.3

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for Scientific Research (C)

    Awarding organization:Japan Society for the Promotion of Science

    TANAKA Takaaki, KAWASE Tomoyuki

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    Grant amount:\5330000 ( Direct Cost: \4100000 、 Indirect Cost:\1230000 )

    Biodegradable separation membrane and scaffold materials have been developed by controlling their internal and surface structures with the aid of surfactants. Asymmetric microporous separation membranes of poly(L-lactic acid) with pores in the order of 0.1 μm were prepared from the polymer solutions containing surfactants with the hydrophilic-lipophilic balance (HLB) values from 15.3 to 16.7. Osteoblast-like cells grew in the 500 μm-thick biodegradable microporous membranes when the cells were seeded on the rough side of the membranes.

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  • Control of composition and morphology of hollow hydroxyapatite microspheres and their application to dental implant materials releasing an antimicrobial agent

    Grant number:21560712

    2009 - 2011

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for Scientific Research (C)

    Awarding organization:Japan Society for the Promotion of Science

    KIMURA Isao, KANATANI Mitsugu, TANAKA Takaaki, AKAZAWA Toshiyuki

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    Grant amount:\4680000 ( Direct Cost: \3600000 、 Indirect Cost:\1080000 )

    Hollow microspheres of calcium phosphate compounds(such as hydroxyapatite, calcium hydrogen phosphate, etc.) were prepared by using multiple emulsions. The composition, morphology, and crystallinity were controlled by the addition of one of a magnesium salt, a carbonate, and low molecular weight organic compounds. For obtaining fundamental knowledge to apply the microspheres to dental implants which release an antimicrobial agent, the conditions for making them adhere onto titanium were investigated. A model implant was fabricated through the following procedures : the formation of microsphere layers on the surface of a titanium screw, the impregnation of a dye solution as the model material of an antimicrobial agent into the microsphere layer, and the coating with biodegradable polymer, poly(lactic acid). With the aim of applying the microspheres to a root canal treatment, composite microcapsules of the microspheres with biodegradable polymer were prepared.

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  • Development of environmental-benign bioseparation membranes by hybridizing biodegradable porous materials and nano inorganic microcapsules

    Grant number:21560807

    2009 - 2011

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for Scientific Research (C)

    Awarding organization:Japan Society for the Promotion of Science

    TANAKA Takaaki, KIMURA Isao

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    Grant amount:\4680000 ( Direct Cost: \3600000 、 Indirect Cost:\1080000 )

    Environmentally benign separation membranes have developed by hybridizing microporous membranes of biodegradable plastics and hydroxyapatite nanoparticles. The depth filter microfiltration membranes of poly(L-lactic acid) developed by Dr. Tanaka, the head of the research project, retained the microcapsules comprised of hydroxyapatite nanoparticles developed by Dr. Kimura, the co-researcher. The hybrid membrane of the biodegradable membrane and hydroxyapatite microcapsules adsorbed bovine serum albumin, a model protein, at a low ionic strength and eluted at an elevated ionic strength.

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  • Control of internal structure of porous membranes of biodegradable polymer blends and its application to bioprocesses

    Grant number:18560748

    2006 - 2008

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for Scientific Research (C)

    Awarding organization:Japan Society for the Promotion of Science

    TANAKA Takaaki, TANIGUCHI Masayuki

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    Grant amount:\3740000 ( Direct Cost: \3200000 、 Indirect Cost:\540000 )

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  • Acceleration of bone formation on mandibular distraction osteogenesis

    Grant number:18592169

    2006 - 2008

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for Scientific Research (C)

    Awarding organization:Japan Society for the Promotion of Science

    KOBAYASHI Tadaharu, TANAKA Takaaki, IZUMI Naoya, EJIRI Sadakazu

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    Grant amount:\4160000 ( Direct Cost: \3500000 、 Indirect Cost:\660000 )

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  • Development of porous membranes of poly (lactic acid) via thermally induced phase separation and its application to bioprocesses

    Grant number:15560673

    2003 - 2005

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for Scientific Research (C)

    Awarding organization:Japan Society for the Promotion of Science

    TANAKA Takaaki, TANIGUCHI Masayuki, AOKI Toshiki, MATSUYMA Hideto

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    Grant amount:\3600000 ( Direct Cost: \3600000 )

    Biodegradable polyesters, e.g., poly(_L-lactic acid), which are degraded by microorganisms in natural environment and degradable in human bodies, have recently received increased attention in bioengineering. In this study we have developed microporous membranes of biodegradable polyesters via thermally induced phase separation. At first effect of characteristics of polymers and conditions of phase separation (diluent, polymer concentration, and thermal history) on the growth of the droplet in phase separation and the structures and pore sizes of the formed porous materials. Then, we made microporous membranes of poly(_L-lactic acid), poly(ε-caprolactone), and their blends. The poly(_L-lactic acid) membrane showed the characteristics of a screen filter in the filtration of yeast cell suspensions. The membrane of a polymer blend of poly(_L-lactic acid) and poly(ε-caprolactone) showed that of a depth filter. The filtration flux with the latter membrane was much higher than that with the former. We also applied poly(_L-lactic acid) membrane to the production of an oxidase, laccase, by membrane-surface liquid culture of white rot fungus, Trametes versicolor. The membrane was stable at 28℃ and the fungus grown on the membrane secreted much more enzyme in the repeated use of membrane-surface liquid culture than in liquid surface culture. The porous biodegradable membranes would contribute to the development of sustainable bioindustry and biomedical engineering.

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  • 生分解性高分子の応用

    2002

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    Grant type:Competitive

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  • Formation and application of microporous biodegradable polyester membranes

    2002

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    Grant type:Competitive

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  • Mechanism and Optimization of Diffusion controlled enantioselective permeable membrane

    Grant number:13555213

    2001 - 2003

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for Scientific Research (B)

    Awarding organization:Japan Society for the Promotion of Science

    AOKI Toshiki, TACHIBANA Kozo, KANEKO Takashi, TANAKA Takaaki

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    Grant amount:\7200000 ( Direct Cost: \7200000 )

    We have synthesized new well-defined polymers and developed optical resolution membranes which have high enantioselevtivity from these polymers. We proposed the separations occurred in diffusion process. However, the reason and detail were not clear and their permeation rates were low. In this study, several optimizations were carried out in these membrane process. For example, molecular structures, conditions in membrane preparation and permeation were examined.
    First, several new optically active polymers were synthesized. Especially, we have discovered new polymerization method to obtain chiral helical polymers. In addition, new chiral helical polymers were prepared by condensation polymerization.
    Secondly, we confirmed the separation of these membrane process occurred in diffusion process. In addition, chiral helicity was found to be effective for enantioselective permeation. In particular, we found that polymeric membranes whose chiral groups had been removed by reaction in membrane state showed enantioselective permeability.
    Lastly, thin membranes such as composite membranes and asymmetric membranes were prepared. High permeation was realized by using these thin membranes. Pervaporation was also tried to make the rate higher.
    In conclusion, we attained higher permeability and obtained many findings for practical use.

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  • Conversion of Agricultural Wastes to Lactic Acid by a Combination of Pentose-fermentative Lactic Acid Bacteria and Reversibly Soluble-insoluble Enzymes

    Grant number:13836003

    2001 - 2002

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for Scientific Research (C)

    Awarding organization:Japan Society for the Promotion of Science

    TANIGUCHI Masayuki, TANAKA Takaaki

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    Grant amount:\3600000 ( Direct Cost: \3600000 )

    1) Glucose and xylose were mainly obtained from delignified rice straw by hydrolysis using cellulase preparation. Glucose and arabinose were mainly obtained from defatted rice bran by hydrolysis using amylase and cellulase preparations.
    2) Lactobacillus vaccinostericus, Lactobacillus pentosus, Pediococcus pentosaceus, and Enterococcus casseliflavus were selected as a xylose-fermented lactic acid bacterium. More lactic acid was produced from MRS medium containing xylose by E. casseliflavus as compared with other lactic acid bacteria.
    3) The amount of lactic acid produced did not increase in a fermentation using MRS medium containing xylose and glucose as a carbon source and E. casseliflavus.
    4) The maintenance of pH at a low value in a simultaneous saccharification and fermentation of non-sterilized rice bran allowed the selective growth of Lactobacillus rhamnosus, a lactic acid bacterium added. About 30 g/L of L-lactic acid was produced from defatted rice bran in a simultaneous saccharification and fermentation in which the pH was kept at 4.5.

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  • Development of Nanobiore actor for Remediation of Foods and Environments

    Grant number:13558072

    2001 - 2002

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for Scientific Research (B)

    Awarding organization:Japan Society for the Promotion of Science

    FUJII Tomoyuki, MIYAWAKI Osato, MIYAKE Noriko, HATTORI Yoshio, TANAKA Takaaki

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    Grant amount:\6900000 ( Direct Cost: \6900000 )

    A thin nanoporous silica membrane was prepared by the two-step sol-gel method for the purpose that the generation of an aggregate having the higher fractal dimension was suppressed in the intermediate process. It succeeded in the preparation of the thin silica film with nanospace of about 3.3nm. The model reaction in which sea sand was made to adsorb the alkylphenol was carried out, and it was found that adsorbing nonylphenol and octylphenol and bisphenol A are quickly decomposed by laccase. The decomposition of an artificial estrogen also confirmed being possible. Log P, which was an useful parameter when we evaluate the solvent property of supercritical carbon dioxide, was estimated. As the pressure of carbon dioxide was increase on the pressure of 3.0 11.8 MPa, log P tended to increase from 0.9 to 2.0. It was shown that the polarity had appeared in carbon dioxide under the supercritical state by the pressure, and that the polarity became small with the increase in the pressure. On the esterification reaction between stearic acid and ethanol using lipase, the reaction rate rose with the increase in the pressure, and it agreed with the increase tendency of log P approximately. It was shown from these results that the solvent property as a bulk had to be considered in addition to the interaction between solute and carbon dioxide, enzyme surface, wall surface, respectively, when it examined the enzyme reaction in nanospace by using supercritical carbon dioxide as a solvent.

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  • 生体触媒を用いた内分泌攪乱化学物質分解システムの開発

    Grant number:12750698

    2000 - 2001

    System name:科学研究費助成事業

    Research category:奨励研究(A)

    Awarding organization:日本学術振興会

    田中 孝明

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    Grant amount:\1800000 ( Direct Cost: \1800000 )

    平成12年度に引き続き,生体触媒による内分泌撹乱化学物質の分解処理システム(バイオレメディエーション・システム)の研究を進めた。平成12年度に開発した回転型バイオリアクターを用いて,砂質に付着したノニルフェノールの分解を行い,反応条件の検討とリスク削減の検証を行った。生体触媒としては白色腐朽菌Trametes sp.由来のラッカーゼを用い,モデル汚染底質としてノニルフェノールを付着させた海砂を用いた。反応装置としては5x5x6cmの正方柱型の反応容器内でモデル汚染底質を回転させる,回転型リアクターを用いた。リアクター中にて,主としてpH5, 30℃にて,15gのモデル汚染底質を酵素溶液45mlを用いて処理した。まず,リアクターの回転軸の角度が反応速度に与える影響について検討した。40-90度の範囲では角度を小さくすると処理速度が高くなったため,40度が最適であった。回転速度については0-45rpmの範囲では10rpmが最適であり,10rpm以上では,回転に要する動力は高くなるが,処理速度は高くならなかった。この傾向は酵素液量を変化させたときも同じであった。次に酵素濃度を変化させて処理反応を行ったところ,反応速度は酵素濃度の約1/2乗に比例してしか増加しなかった。砂粒子の表面上のノニルフェノールに対しては,酵素濃度を高めても酵素濃度当りの作用効率が低下すると考えられた。反応温度を高めると40℃までは反応速度が高められた。ただし,10℃においても反応時間を長くすると90%以上の処理が可能なことが示された。最後に,オスメダカを用いた暴露実験にて,処理後の砂質からの抽出物の毒性評価を,血中ビテロゲニン濃度を指標として行った。その結果,ラッカーゼ処理によりノニルフェノールの内分泌撹乱作用が大きく低下することが示された。

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  • 白色腐朽菌のリグニン分解酵素を用いた酸化重合反応による新規性分解性ポリマーの合成

    Grant number:11875175

    1999

    System name:科学研究費助成事業

    Research category:萌芽的研究

    Awarding organization:日本学術振興会

    谷口 正之, 田中 孝明

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    Grant amount:\2100000 ( Direct Cost: \2100000 )

    本研究では,ラッカーゼを用いてフェノール類から生分解性ポリマーを合成することについて検討した。この研究において、以下の結果を得た。
    1)バイオリアクターを用いてCoriolus versicolorによるラッカーゼの効率的生産について検討した結果、固体培地の初期水分含量を70%とした培養14日目に最大酵素活性が得られた。
    2)固体培地から回収したラッカーゼを,硫安沈殿で濃縮した後、イオン交換クロマトグラフィーと疎水性クロマトグラフィーによって精製した結果、比活性が45倍となった高純度の酵素を得ることができた。
    3)精製したラッカーゼを用いて、フェノールおよびフェニルフェノールを基質モノマーとして重合反応を行った結果、いずれもpHが4.5のときにポリマーの収率が最大となった。
    4)フェニルフェノールを基質モノマーとして用いた場合には、反応溶媒にアセトンを添加することによってポリマー収率が向上した。アセトンを30%添加したときのポリマー収率は98%であった。
    5)フェノールを基質モノマーとして重合反応を行った場合に生成したポリマーは、ジメチルホルムアミドに溶解しない高分子量であった。一方、フェニルフェノールを重合した場合に生成したポリマーは、ジメチルホルムアミドに溶解し、その分子量は6,000-10,000であった。
    6)フェノールおよびフェニルフェノールの酵素酸化重合物の熱重量分析を行った結果、これらの重合物は340℃まで安定であった。

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  • Production of Bifidogenic Growth Stimulator and Its Application to Development of Probiotics

    Grant number:10555286

    1998 - 2000

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for Scientific Research (B).

    Awarding organization:Japan Society for the Promotion of Science

    TANIGUCHI Masayuki, TANAKA Takaaki, IIDA Takamitsu, HOSHINO Kazuhiro, HOJO Kenichi, KANEKO Tsutomu

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    Grant amount:\12700000 ( Direct Cost: \12700000 )

    1. The concentration and the productivity per time of bifidogenic growth stimulator were increased by using a co-culture system for a homolactic acid bacterium and a propionic acid bacterium, a culture system with microfiltration, a fed-batch culture system with feed back control of carbon source concentration, and a culture system with microfiltration and feed back control of carbon source concentration.
    2. A novel bioreactor system with two fermentors and microfiltration modules was developed for efficient production bifidobacterial cells by a co-culture of Propionibacterium freudenreichii and Bifidobacterium adolescentis as well as clarification of interactions between the two bacteria.
    3. The proposed co-culture system allowed the successful exchange of culture medium between two fermentors. When P.freudenreichii and B.adolescentis, B.longum, or B.breve were cultivated individually in the new co-culture system, the amount of bifidobacterial cells produced was 1.2-1.6 fold as high as that with batch culture.

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  • 残留農薬迅速分解のための固定化リグニン分解酵素バイオリアクターシステムの開発

    Grant number:09750869

    1997 - 1998

    System name:科学研究費助成事業

    Research category:奨励研究(A)

    Awarding organization:日本学術振興会

    田中 孝明

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    Grant amount:\2100000 ( Direct Cost: \2100000 )

    リグニン分解酵素を用いた残留農薬の迅速分解を行うためのバイオリアクターシステムの開発を目的として、その基礎となる研究を行った。まず、酵素の生産条件を検討した。リグニン分解酵素生産菌としては4種類の担子菌(Corilus versicolor ,Pleurotus osteatus,Phanerochaete chrysosporium,Bjerkandera adusta)を用いた。固体培養と液体培養とでリグニン分解酵素(リグニンペルオキシダーゼ、マンガンペルオキシダーゼ、ラッカーゼ)の培養体積当たりの生産量を比較したところ、固体培養の酵素生産量が液体培養の場合と比較して10から100倍程度高かった。次に、固体培養の菌体外酵素液を用いて農薬の一種ペンタクロロフェノールの分解実験を行った。酵素としては主としてC.versicolorの菌体外酵素を用いた。0.1-1.0mMのペンタクロロフェノール溶液(10%メタノール含有)に酵素液を添加したところ、7日間で80%分解された。各種農薬の水への溶解度を検討したところ、極めて低かった。そこで河川底質に残留する農薬の酵素分解を検討した。汚染底質のモデル系として海砂にアルキルフェノールを吸着させたものを用いた。アルキルフェノールは多くの農薬と同じく疎水性であり、近年、一部の農薬と同様に内分泌撹乱化学物質として環境中での毒性が懸念されている。C.versicolorの菌体外酵素液でアルキルフェノールは分解された。また、酵素分解の至適pHは5であった。また、C.versicolorの菌体外酵素液から精製したラッカーゼでも海砂に吸着したアルキルフェノールが分解できた。ラッカーゼは反応に酸素を要求することから、汚染底質を撹拌しながら分解反応を行うバイオリアクターについて考察した。

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  • リグニン分解酵素の応用

    1996

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    Grant type:Competitive

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  • Application of lignin degrading enzymes

    1996

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    Grant type:Competitive

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  • DEVELOPMENT OF BIOREACTOR SYSTEM FOR OLIGOPEPTIDE SYNTHESIS COMBINED WITH A SIMULATED MOVING BED ADSORBER

    Grant number:07555563

    1995 - 1996

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for Scientific Research (A)

    Awarding organization:Japan Society for the Promotion of Science

    NAKANISHI Kazuhiro, SHIOTA Katashi, UTAGAWA Takashi, SIRAI Yoshihito, TANAKA Takaaki, SAKIYAMA Takaharu

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    Grant amount:\1700000 ( Direct Cost: \1700000 )

    In this study, we aimed to construct an effective bioreactor for synthesis of aspartame precursor, as a model oligopeptide in an organic solvent, using thermolysin immobilized onto Amberlite XAD-7. First, we investigated the factors affecting the synthesis of the asparatme precursor using an immobilized enzyme. We studied a synthetic rate, particularly in a mixed organic solvent with ethyl acetate and tert-amyl alcohol. Using a mixed solvent of 33% tert-amyl alcohol and 67% ethyl acetate, the activity and stability of the immobilized thermolysin were high enough to conduct a continuous reaction. With increasing the content of tert-amyl alcohol, the stability of the immobilized enzyme was enhanced. The reason for the high activity and stability of the immobilized thermolysin in the presence of tert-amyl alcohol was studied from the viepoint of stability of calcium bound on the enzyme molecule, a stabilizing factor for thermolysin. An optimum water content for the synthesis of asparatame precursor was found to be 4-6 % in the mixed organic solvent. The concentration of L-phenyllalanine methyl ester (PheOMe) and N- (benzy loxy carbonyl) -L-as partic acid (Z-Asp) was optimized. The concentration of PheOMe was set to 200 mM and that of Z-Asp was 40 mM.We could also conduct a continuous reaction in a column reactor at 40゚C.The conversion at the outlet of the reactor with respect of Z-Asp was around 99% with a space velocity of 4.9 (1/h). Based on this result, we aimed to develop a bioreactor system combined with a simulated moving bed absorber. Namely, at the outlet of the bioreactor, the Z-Asp concentration was negligible low with asparatame precursor Z-Asp PheOMe of around 40 mM and PheOMe of 160 mM.Thus, we could recycle PheOMe to the bioreactor with continuous separation of PheOMe and Z-AspPheOMe using a simulated mooving bed adsorber. We found Dowex MWA-1 as a suitable adsorbent and analyzed adsorption/desorption behaviors of PheOMe and Z-AspPheOMe. We also calculated a separation performance for a simulated moving bed adosorber. We also showed a possible bioreactor system which could be used for an industrial production. Finally, we briefly compared an enzymatic synthesis and chemical synthesis.

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  • 食品加工プロセスにおける汚れの洗浄過程の解析とその効率化

    Grant number:06650859

    1994

    System name:科学研究費助成事業

    Research category:一般研究(C)

    Awarding organization:日本学術振興会

    中西 一弘, 田中 孝明, 崎山 高明

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    Grant amount:\2000000 ( Direct Cost: \2000000 )

    食品製造工程やバイオプロセスにおいては、必ず装置表面に付着残存する汚れ成分を除去するための洗浄操作が実施されている。本研究では、洗浄操作の最適化を最終目的として、その基礎となる溶質のステンレス表面への付着・脱離現象の解明を行った。モデル汚れ成分として、β-ラクトグロブリンを取り上げた。溶質の付着特性と洗浄特性を精度よく調べるために、比表面積の大きなステンレス微粒子を用いた。タンパク質の付着量に及ぼす、付着時間、pH、温度、蛋白濃度の影響を調べた。温度が50℃以下では、その最大付着量は単分子層が形成される量にほぼ一致したが、β-ラクトグロブリンの変性温度である65℃前後から急激に増加し、単分子層の数倍程度となった。付着量はpHに強く依存した。β-ラクトグロブリンの等電点である5.1以下では付着量は急激に増加したが、これは負に帯電しているステンレス表面と正に帯電しているタンパク質との間の静電的相互作用によるものであることが判明した。また、未変性タンパク、予め変性したタンパク、及びSH基を修飾したタンパク質を用いて吸着実験を行った結果、β-ラクトグロブリンは、静電気的相互作用、及び疎水結合によりステンレス表面に付着し、その後タンパク質間でS-S結合の形成や交換反応により付着が進行することが明らかになった。
    洗浄特性に関しては、申請者らが開発した汚れの付着したステンレス粒子充填カラムを用いるステップ応答法を用いた。アルカリ洗浄と、プロテアーゼを用いる酵素洗浄に着目して比較検討した。酵素法では、アルカリ洗浄に比較してより低いpHで、洗浄が可能であり、しかも残存付着量も少ないことが判明した。また、アルカリ洗浄ではタンパク分子、あるいはその凝集体の形で、一方、酵素洗浄では分解ペプチドの形で、表面から脱離することが示唆された。

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  • 培地由来の微粒子を含む培養液のクロスフロー膜濾過に関する基礎的研究

    Grant number:05750719

    1993

    System name:科学研究費助成事業

    Research category:奨励研究(A)

    Awarding organization:日本学術振興会

    田中 孝明

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    Grant amount:\800000 ( Direct Cost: \800000 )

    本研究は微粒子を含む培地で培養した菌体培養液のクロスフロー濾過特性を明らかにすることを目的として行った。平成5年度に得られた研究成果を以下に示す。
    (1)微粒子が混入した糖蜜を培地成分としてパン酵素を培養した培養液をクロスフロー濾過したときの濾過速度の変化を濾過期間を菌体層形成過程と微粒子層形成過程に分けることにより予測できることを示した。走査電子顕微鏡観察により微粒子の堆積を実証できた。クロスフロー速度、圧力などを変化させたときに濾過特性がどのように変化するかについてもまとめることができた。特にクロスフロー速度を高めた場合、通常の粒子懸濁液のクロスフロー濾過においては濾過速度が高くなるのに対して、微粒子混入系においては濾過速度が低下することを発見し、上述の2段階付着層形成過程モデルによりも導けることを示した。さらに逆洗を組み合わせたときの操作条件の最適化についてもまとめた。これらの結果は、国際食品工学会議(平成5年6月、千葉)、化学工学会秋季大会(平成5年10月、京都)にて発表した。
    (2)微粒子を含む培養液のモデル系として粒径の異なる高分子ラテックスを合成し、その濾過特性を調べた。デッドエンド型濾過特性およびクロスフロー濾過特性を調べた。これと菌体の濾過特性を組み合わせてクロスフロー濾過における濾過速度の粒径依存性についてまとめた。さらに菌体とラテックスの混合懸濁液を調製しクロスフロー濾過特性を検討した。

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  • 微生物菌体分離のためのクロスフロー瀘過法の高効率化

    Grant number:04203223

    1992

    System name:科学研究費助成事業

    Research category:重点領域研究

    Awarding organization:日本学術振興会

    中西 一弘, 田中 孝明, 富田 憲史

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    Grant amount:\2500000 ( Direct Cost: \2500000 )

    バイオプロセスにおける除菌・集菌工程の効率化を目的として、クロスフロー膜瀘(Cross Flow Filtration;CFF)に着目して、その膜瀘過流束に及ぼす諸因子の解明を行うと共に、膜透過流束を高くするための方策を検討した。具体例として、(1)パン酵母培養液と、(2)プルラン培養液のCFFを取り上げた。
    YPD培地で純粋培養したパン酵母培養液のCFFにおいては、定常流束だけではなく、定常状態に達するまでの流束変化も瀘過理論から推測される結果と一致した。すなわち、クロスフロー膜瀘過時におけるケーク層の充填状態はデッドエンド型瀘過の場合と同様であることが判明した。膜透過流束に及ぼす膜間圧力差、循環流速、菌体濃度、膜の種類などの影響を調べた。循環流速と菌体濃度に関しては、モジュール内部が菌体により閉塞を起こす限界の値が存在することが判明した。
    天然培地であるモラッセスを用いて培養を行った場合の膜透過流束は、YPD培地で培養したパン酵母懸濁液の結果と比較して著しく低下した。この理由として、モラッセス中に存在する微粒子が菌体ケーク層表面上に薄い層をなして積もり、しかもこの薄層の比抵抗が菌体ケーク層よりも著しく高いことが明らかにされた。膜透過流束を高めるための方策を検討したところ、逆洗が最も効果的であった。逆洗による膜透過流束の回復は膜孔径に依存し、膜孔径が0.8μm以下の膜では殆どみられず、孔径が3μmあるいは5μmの膜では膜透過流束の回復は100%であった。逆洗を行うことにより、膜透過流束は5倍以上増加した。
    プルラン培養液のクロスフロー瀘過においては、菌体の形状が膜透過流束に大きく影響することが判明した。培養条件を工夫することにより膜透過流束は数十倍に増加した。

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  • Membrane filtration of microbial suspension

    1990

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    Grant type:Competitive

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  • 菌体懸濁液のクロスフロー濾過

    1990

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    Grant type:Competitive

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  • 微生物菌体分離のためのクロスフロ-濾過法の高効率化

    Grant number:02203238

    1990

    System name:科学研究費助成事業

    Research category:重点領域研究

    Awarding organization:日本学術振興会

    中西 一弘, 田中 孝明, 富田 憲史

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    Grant amount:\3000000 ( Direct Cost: \3000000 )

    本年度は以下の三点の項目に関して検討を行った。
    (1)パン酵母懸濁液のクロスフロ-濾過機構の解明
    薄層流路型クロスフロ-濾過装置を用いて、パン酵母懸濁液のクロスフロ-濾過を行い、その膜透過流束に及ぼす諸因子を調べた。これらの実験結果から、膜透過機構に関して検討を行ったところ以下の知見が得られた。濾過開始直後においては、きわめて短期間であるが、菌体はDeadーend型の濾過の場合と同様な充填構造を取りながらも膜面上に堆積する。その後、菌体付着層表面近傍において、内部よりも密な構造の付着層が形成されることが明らかにされた。この理由として循環中に一部の菌体が変形しやすい構造に変質するためであることが示唆された。
    (2)膜透過流束を高くするための新規な操作方法の開発
    高い膜透過流束を得る目的で、種々の操作方法を検討した結果、濾口を周期的に開閉し、しかも濾口を閉じた時に気泡を供給すると、膜透過流束が数倍から10倍増加することが判明した。最適な条件では、2%のパン酵母懸濁液を10%に濃縮した場合の平均膜透過流束は3501/h・m^2に達した。
    (3)高粘性培養液のクロスフロ-濾過
    微生物菌体の中には、多糖など高分子物質を分泌するものが少なくない。これらの高分子物質は食品添加剤や生理活性物質としての理用が注目されている。本研究では、ケ-ススタディ-として、<Aureobasidium>___ー <pullulanns>___ーによる多糖プルランの発酵生産を行い、その高粘性培養液からの菌体の回収をクロスフロ-濾過法により行った。膜透過流束は低に値であったが、濾液は完全に清澄であり、プルランの膜透過率も90%と高く、クロスフロ-濾過法の有用性が示された。

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  • Syntheses of Umami Peptides with Immobilized Papain in Water-Immiscible Organic Solvent Systems

    Grant number:01560151

    1989 - 1990

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for General Scientific Research (C)

    Awarding organization:Japan Society for the Promotion of Science

    NAKANISHI Kazuhiro, TANAKA Takaaki

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    Grant amount:\2200000 ( Direct Cost: \2200000 )

    Dipeptides such as L-glutamic acid dimer and L-glutamic acid dimer are known as a substance posessing umami (brothy) taste and a bitter taste masking effect. In this study, we attempted to synthesize precursors of four peptides, i. e., GluGlu, GluAsp, AspGlu, and AspAsp with an immobilized papain in organic solvent systems. As an acid component of a substrate, N-(benzyloxycarbonyl)-L-glutamic acid and N-(benzyloxycarbonyl)-L-aspartic acid, and as an amine component L-glutanic acid diethyl ester and L-aspartic acid diethyl ester were used. In most cases, we focussed on the synthesis of N-(benzyloxycarbonyl)-L-glutamyl-L-glutamic acid diethyl ester (Z-GluGlu(OEt)_2) at 30 ^゚C.
    The results obtained are Summarized as follows :
    1) The synthetic reaction did not take place in buffer solution.
    2) In the aqueous-organic biphasic reaction systems with ethyl acetate as an organic solvent, the maximum yield of Z-GluGlu (OEt) _2 reached about 70%.
    3) The enzyme was immobilized onto various porous supports. The enzyme was first adsorbed onto the support and then cross-linked with glutaraldehyde. The highest activity was obtained when the enzyme was adsorbed onto porous ceramic support (SM10-C1, Nippon Gaishi K. K.).
    4) The activity of immobilized enzyme depended strongly on the water content in an outer organic solvent. The highest activity was obtained at a water content of 2.5% with the enzyme immobilized onto SM10-C1.
    5) L-Glutamic acid diethyl ester, an amine component of the substrate was non-enzymatically coverted to L-pyroglutamic acid diethyl ester. To reduce the effect of non-enzymatic reaction, the increase of the concentration of the immobilized enzyme was found most effective. With 100 mM Z-Glu and 200 mM Glu (OEt) _2, and immobilized enzyme concentration of 8%, the yield became higher than 90%.
    6) The other three dipeptides precursors were also obtained with quite a high yield under the same reaction conditions as shown in (5).
    7) The products were identified by analyzing with nuclear magnetic resonance, mass spectrum, and infrared adsorption spectrum.

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  • 生物分離工学

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    Grant type:Competitive

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  • 酵素利用技術

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    Grant type:Competitive

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  • 膜分離技術(特に精密濾過)

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    Grant type:Competitive

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Teaching Experience

  • 国際工学概論

    2024
    Institution name:新潟大学

  • 統合化学入門

    2022
    Institution name:新潟大学

  • インターンシップ

    2022
    Institution name:新潟大学

  • 機能材料工学実験II

    2021
    Institution name:新潟大学

  • リメディアル演習

    2021
    Institution name:新潟大学

  • 生物機能材料科学

    2021
    Institution name:新潟大学

  • 論文輪講II

    2020
    Institution name:新潟大学

  • 技術英語I

    2020
    Institution name:新潟大学

  • 技術英語II

    2020
    Institution name:新潟大学

  • 論文輪講I

    2020
    Institution name:新潟大学

  • 先端科学技術総論

    2020
    Institution name:新潟大学

  • 材料科学実験II

    2019
    Institution name:新潟大学

  • 総合工学概論

    2017
    Institution name:新潟大学

  • 工学リテラシー入門(化学材料分野)

    2017
    Institution name:新潟大学

  • 材料科学概論

    2017
    Institution name:新潟大学

  • 基礎有機化学

    2016
    Institution name:新潟大学

  • 生物機能材料科学

    2015
    -
    2021
    Institution name:新潟大学

  • 材料生産システム博士特定研究Ⅱ

    2015
    Institution name:新潟大学

  • 中間発表

    2015
    Institution name:新潟大学

  • 材料生産システム博士セミナーⅡ

    2015
    Institution name:新潟大学

  • 情報機器操作入門

    2015
    Institution name:新潟大学

  • 外国語論文解説・討論Ⅱ

    2015
    Institution name:新潟大学

  • 機能材料化学概論

    2014
    -
    2016
    Institution name:新潟大学

  • 外国語論文解説・討論Ⅰ

    2014
    Institution name:新潟大学

  • 機能材料科学コース演習

    2014
    Institution name:新潟大学

  • 技術英語入門

    2014
    Institution name:新潟大学

  • 技術者倫理

    2014
    Institution name:新潟大学

  • 材料生産システム博士特定研究Ⅰ

    2014
    Institution name:新潟大学

  • 材料生産システム博士セミナーⅠ

    2014
    Institution name:新潟大学

  • 自然科学総論Ⅱ

    2013
    -
    2020
    Institution name:新潟大学

  • 機能材料科学文献詳読Ⅱ

    2012
    -
    2015
    Institution name:新潟大学

  • 機能材料科学セミナーⅡ

    2012
    -
    2015
    Institution name:新潟大学

  • 材料生産システム特定研究Ⅱ

    2012
    -
    2015
    Institution name:新潟大学

  • 機能材料科学演習

    2012
    -
    2015
    Institution name:新潟大学

  • 機能材料科学文献詳読Ⅰ

    2012
    -
    2015
    Institution name:新潟大学

  • 機能材料科学セミナーⅠ

    2012
    -
    2015
    Institution name:新潟大学

  • 材料生産システム特定研究Ⅰ

    2012
    -
    2015
    Institution name:新潟大学

  • 研究発表演習・発表

    2012
    -
    2015
    Institution name:新潟大学

  • 卒業研修

    2009
    Institution name:新潟大学

  • 機能材料科学総論Ⅱ

    2009
    Institution name:新潟大学

  • 工学リテラシー入門(機能材料工学科)

    2009
    -
    2016
    Institution name:新潟大学

  • 最先端技術を支える化学 II

    2008
    -
    2013
    Institution name:新潟大学

  • 機能材料工学実験IV

    2008
    -
    2010
    Institution name:新潟大学

  • スタディスキルズ(機能材料工学)

    2008
    Institution name:新潟大学

  • 卒業研究

    2007
    Institution name:新潟大学

  • 生物材料工学

    2007
    Institution name:新潟大学

  • 生物反応プロセス工学

    2007
    Institution name:新潟大学

  • 論文輪講

    2007
    -
    2022
    Institution name:新潟大学

  • 技術英語

    2007
    -
    2022
    Institution name:新潟大学

  • 機能材料工学実験III

    2007
    -
    2018
    Institution name:新潟大学

  • 基礎物理化学

    2007
    -
    2015
    Institution name:新潟大学

  • 化学実験

    2007
    -
    2014
    Institution name:新潟大学

  • 生物素材工学特論

    2007
    -
    2013
    Institution name:新潟大学

  • 卒業基礎研究

    2007
    -
    2008
    Institution name:新潟大学

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