Updated on 2024/03/28

写真a

 
SUGIMOTO Hayuki
 
Organization
Academic Assembly Institute of Science and Technology NOUGAKU KEIRETSU Associate Professor
Faculty of Agriculture Associate Professor
Graduate School of Science and Technology Life and Food Sciences Applied Life and Food Sciences Associate Professor
Title
Associate Professor
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Degree

  • 博士(学術) ( 2008.7   三重大学 )

Research Interests

  • 微生物

  • タンパク質・酵素

  • フォールディング

  • 構造安定性

  • calorimetry

Research Areas

  • Life Science / Applied biochemistry

  • Life Science / Applied microbiology

  • Life Science / Functional biochemistry

Research History (researchmap)

  • Niigata University   Faculty of Agriculture

    2011.4

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  • Kuwansei Univ.   Graduate School of Science and Technology

    2008.8 - 2011.3

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Research History

  • Niigata University   Graduate School of Science and Technology Life and Food Sciences Applied Life and Food Sciences   Associate Professor

    2016.4

  • Niigata University   Graduate School of Science and Technology Life and Food Sciences   Assistant Professor

    2011.4 - 2016.3

  • Niigata University   Graduate School of Science and Technology Life and Food Sciences   Assistant Professor

    2011.4 - 2016.3

  • Niigata University   Abolition organization Applied Biological Chemistry   Assistant Professor

    2011.4 - 2016.3

Education

  • Mie Univ.   Graduate school of Bioresources   博士後期課程

    2005.4 - 2008.7

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Professional Memberships

  • THE BIOPHYSICAL SOCIETY OF JAPAN

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  • THE JAPANESE BIOCHEMICAL SOCIETY

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  • The Protein Society (USA)

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  • PROTEIN SCIENCE SOCIETY OF JAPAN

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  • THE JAPAN SOCIETY OF CALORIMETRY AND THERMAL ANALYSIS

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  • JAPAN SOCIETY FOR BIOSCIENCE, BIOTECHNOLOGY, AND AGROCHEMISTRY

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Papers

  • Chitinase system of Aeromonas salmonicida, and characterization of enzymes involved in chitin degradation Reviewed International journal

    Iuliia Pentekhina, Tatsuyuki Hattori, Dinh Minh Tran, Mizuki Shima, Takeshi Watanabe, Hayuki Sugimoto, Kazushi Suzuki

    Bioscience, Biotechnology, and Biochemistry   84 ( 9 )   1936 - 1947   2020.9

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Informa UK Limited  

    The genes encoding chitin-degrading enzymes in Aeromonas salmonicida SWSY-1.411 were identified and cloned in Escherichia coli. The strain contained two glycoside hydrolase (GH) families 18 chitinases: AsChiA and AsChiB, two GH19 chitinases: AsChiC and AsChiD, and an auxiliary activities family 10 protein, lytic polysaccharide monooxygenase: AsLPMO10A. These enzymes were successfully expressed in E. coli and purified. AsChiB had the highest hydrolytic activity against insoluble chitin. AsChiD had the highest activity against water-soluble chitin. The peroxygenase activity of AsLPMO10A was lower compared to SmLPMO10A from Serratia marcescens. Synergism on powdered chitin degradation was observed when AsChiA and AsLPMO10A were combined with other chitinases of this strain. More than twice the increase of the synergistic effect was observed when powdered chitin was treated by a combination of AsLPMO10A with all chitinases. GH19 chitinases suppressed the hyphal growth of Trichoderma reesei.

    DOI: 10.1080/09168451.2020.1771539

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  • Unfolding of CBP21, a lytic polysaccharide monooxygenase, without dissociation of its copper ion cofactor Reviewed

    Hayuki Sugimoto, Yuichi Nakajima, Ayaka Motoyama, Erina Katagiri, Takeshi Watanabe, Kazushi Suzuki

    Biopolymers   111 ( 1 )   e23339   2020.1

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    Publishing type:Research paper (scientific journal)   Publisher:Wiley  

    DOI: 10.1002/bip.23339

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    Other Link: https://onlinelibrary.wiley.com/doi/full-xml/10.1002/bip.23339

  • A novel chitin-binding mode of the chitin-binding domain of chitinase A1 from Bacillus circulans WL-12 revealed by solid-state NMR Reviewed

    Tanaka, Hiroki, Akutsu, Hideo, Yabuta, Izumi, Hara, Masashi, Sugimoto, Hayuki, Ikegami, Takahisa, Watanabe, Takeshi, Fujiwara, Toshimichi

    FEBS LETTERS   592 ( 18 )   3173 - 3182   2018.9

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    Chitin-binding domain of chitinase A1 (ChBD(chiA1)) is characteristic because it binds only to insoluble crystalline chitin. While binding sites of major carbohydrate-binding modules carry multiple aromatic rings aligned on a surface, lethal mutations for ChBD(chiA1) were reported only at W687, a location completely different from the site mentioned above, in spite of their similar main-chain folds. Here, the structural mechanism underlying its crystalline chitin binding was uncovered by solid-state NMR. Based on C-13-and N-15-signal assignment of microcrystalline ChBD(chiA1), the chemical shift perturbation on chitin binding was carefully examined. The perturbation was greatest at W687 and nonaromatic residues surrounding it, revealing their direct involvement in chitin binding. These residues and Q679 should provide a novel chitin-binding platform parallel to the W687 ring.

    DOI: 10.1002/1873-3468.13226

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  • Identification and characterization of chitinolytic bacteria isolated from a freshwater lake Reviewed

    Dinh Minh Tran, Hayuki Sugimoto, Dzung Anh Nguyen, Takeshi Watanabe, Kazushi Suzuki

    Bioscience, Biotechnology and Biochemistry   82 ( 2 )   343 - 355   2018

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Japan Society for Bioscience Biotechnology and Agrochemistry  

    To develop a novel type of biocontrol agent, we focus on bacteria that are characterized by both chitinase activity and biofilm development. Chitinolytic bacteria were isolated from sediments and chitin flakes immersed in the water of a sand dune lake, Sakata, in Niigata, Japan. Thirty-one isolates from more than 5100 isolated strains were examined chitinase activity and biofilm formation. Phylogenetic analysis of these isolates based on the 16S rRNA gene sequences revealed that most isolates belonged to the family Aeromonadaceae, followed by Paenibacillaceae, Enterobacteriaceae, and Neisseriaceae. The specific activity of chitinase of four selected strains was higher than that of a reference strain. The molecular size of one chitinase produced by Andreprevotia was greater than that of typical bacterial chitinases. The dialyzed culture supernatant containing chitinases of the four strains suppressed hyphal growth of Trichoderma reesei. These results indicate that these four strains are good candidates for biocontrol agents.

    DOI: 10.1080/09168451.2017.1422969

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  • Calorimetric study on the binding of beta-cyclodextrin to the starch-binding domain of Aspergillus niger glucoamylase Reviewed

    Masuda Y, Miyake H, Tanaka A, Sugimoto H

    Netsu Sokutei   44 ( 4 )   135 - 138   2017

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  • Differences in the roles of the two surface-exposed tyrosine residues, Y240 and Y481, of Serratia marcescens chitinase B during processive degradation of crystalline chitin Reviewed

    Hayuki Sugimoto, Keita Nakamura, Yuji Nishino, Yuta Idezawa, Akiko Fujinuma, Kazushi Suzuki, Takeshi Watanabe

    JOURNAL OF GENERAL AND APPLIED MICROBIOLOGY   61 ( 6 )   255 - 261   2016

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:MICROBIOL RES FOUNDATION  

    Chitinase B from Serratia marcescens 2170 is one of the processive chitinases, and it has a linear path of aromatic amino acid residues on the surface and in the catalytic cleft. There are four surface-exposed residues lined-up towards the cleft, Y481, W479, W252, and Y240. The substitution of these residues with alanine causes a decrease in both the extent of the substrate binding and the hydrolytic activity (Katouno et al., 2004). Here, we examine the three mutants without losing the substrate-binding ability, Y240W, Y481W, and Y240W/Y481W. These mutants were prepared for a detailed analysis of the functions of Y240 and Y481, which showed a lower contribution to substrate binding than W479 and W252. The parameters for the binding of the three mutants to crystalline beta-chitin were similar to those for the wild type. The hydrolytic activity of Y240W and Y240W/Y481W against crystalline beta-chitin was significantly decreased. However, the hydrolytic activity of Y481W was similar to that of the wild type, indicating some differences in the roles of Y240 and Y481 during the processive degradation of crystalline beta-chitin. Taken together with the previous results, it was suggested that while Y240 and Y481 were required for the substrate binding, Y240 had additional roles in the processive degradation of crystalline beta-chitin, possibly in guiding a chitin chain into the catalytic cleft.

    DOI: 10.2323/jgam.61.255

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  • Analysis of subfamily C chitinases in bacterial glycoside hydrolase family 18 Reviewed

    21 ( 3 )   209 - 218   2015.11

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    Language:Japanese   Publishing type:Research paper (scientific journal)  

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  • Two-way traffic of glycoside hydrolase family 18 processive chitinases on crystalline chitin Reviewed

    Kiyohiko Igarashi, Takayuki Uchihashi, Taku Uchiyama, Hayuki Sugimoto, Masahisa Wada, Kazushi Suzuki, Shohei Sakuda, Toshio Ando, Takeshi Watanabe, Masahiro Samejima

    NATURE COMMUNICATIONS   5   3975   2014.6

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    Processivity refers to the ability of synthesizing, modifying and degrading enzymes to catalyse multiple successive cycles of reaction with polymeric substrates without disengaging from the substrates. Since biomass polysaccharides, such as chitin and cellulose, often form recalcitrant crystalline regions, their degradation is highly dependent on the processivity of degrading enzymes. Here we employ high-speed atomic force microscopy to directly visualize the movement of two processive glycoside hydrolase family 18 chitinases (ChiA and ChiB) from the chitinolytic bacterium Serratia marcescens on crystalline beta-chitin. The half-life of processive movement and the velocity of ChiA are larger than those of ChiB, suggesting that asymmetric subsite architecture determines both the direction and the magnitude of processive degradation of crystalline polysaccharides. The directions of processive movements of ChiA and ChiB are observed to be opposite. The molecular mechanism of the two-way traffic is discussed, including a comparison with the processive cellobiohydrolases of the cellulolytic system.

    DOI: 10.1038/ncomms4975

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  • NMR Analysis of a Kinetically Trapped Intermediate of a Disulfide-Deficient Mutant of the Starch-Binding Domain of Glucoamylase Reviewed

    Hayuki Sugimoto, Yasuo Noda, Shin-ichi Segawa

    JOURNAL OF MOLECULAR BIOLOGY   412 ( 2 )   304 - 315   2011.9

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD  

    A thermally unfolded disulfide-deficient mutant of the starch-binding domain of glucoamylase refolds into a kinetically trapped metastable intermediate when subjected to a rapid lowering of temperature. We attempted to characterise this intermediate using multidimensional NMR spectroscopy. The (1)H-(15)N heteronuclear single quantum coherence spectrum after a rapid temperature decrease (the spectrum of the intermediate) showed good chemical shift dispersion but was significantly different from that of the native state, suggesting that the intermediate adopts a nonnative but well-structured conformation. Large chemical shift changes for the backbone amide protons between the native and the intermediate states were observed for residues in the beta-sheet consisting of strands 2, 3, 5, 6, and 7 as well as in the C-terminal region. These residues were found to be in close proximity to aromatic residues, suggesting that the chemical shift changes are mainly due to ring current shifts caused by the aromatic residues. The two-dimensional nuclear Overhauser enhancement (NOE) spectroscopy experiments showed that the intermediate contained substantial, native-like NOE connectivities, although there were fewer cross peaks in the spectrum of the intermediate compared with that of the native state. It was also shown that there were native-like interresidue NOEs for residues buried in the protein, whereas many of the NOE cross peaks were lost for the residues involved in a surface-exposed aromatic cluster. These results suggest that, in the intermediate, the aromatic cluster at the surface is structurally less organised, whereas the interior of the protein has relatively rigid, native-like side-chain packing. (C) 2011 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.jmb.2011.07.025

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  • Kinetically trapped metastable intermediate of a disulfide-deficient mutant of the starch-binding domain of glucoamylase Reviewed

    Hayuki Sugimoto, Miho Nakaura, Shigenori Nishimura, Shuichi Karita, Hideo Miyake, Akiyoshi Tanaka

    PROTEIN SCIENCE   18 ( 8 )   1715 - 1723   2009.8

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JOHN WILEY & SONS INC  

    Refolding of a thermally unfolded disulfide-deficient mutant of the starch-binding domain of glucoamylase was investigated using differential scanning calorimetry, isothermal titration calorimetry, CD, and (1)H NMR. When the protein solution was rapidly cooled from a higher temperature, a kinetic intermediate was formed during refolding. The intermediate was unexpectedly stable compared with typical folding intermediates that have short half-lives. It was shown that this intermediate contained substantial secondary structure and tertiary packing and had the same binding ability with beta-cyclodextrin as the native state, suggesting that the intermediate is highly-ordered and native-like on the whole. These characteristics differ from those of partially folded intermediates such as molten globule states. Far-UV CD spectra showed that the secondary structure was once disrupted during the transition from the intermediate to the native state. These results suggest that the intermediate could be an off-pathway type, possibly a misfolded state, that has to undergo unfolding on its way to the native state.

    DOI: 10.1002/pro.188

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  • Stabilization mechanism of chloride ion on thermal denaturation of Arthrobacter sarcosine oxidase Reviewed

    Sugimoto H, Nishiya Y, Miyake H, Tanaka A

    Netsu Sokutei   35   76 - 80   2008

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  • Thermodynamic effects of disulfide bond on thermal unfolding of the starch-binding domain of Aspergillus niger glucoamylase Reviewed

    Hayuki Sugimoto, Miho Nakaura, Yoshie Kosuge, Kunio Imai, Hideo Miyake, Shuichi Karita, Akiyoshi Tanaka

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   71 ( 6 )   1535 - 1541   2007.6

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:TAYLOR & FRANCIS LTD  

    The thermodynamic effects of the disulfide bond of the fragment protein of the starch-binding domain of Aspergillus niger glucoamylase was investigated by measuring the thermal unfolding of the wild-type protein and its two mutant forms, Cys3Gly/Cys98Gly and Cys3Ser/Cys98Ser. The circular dichroism spectra and the thermodynamic parameters of binding with beta-cyclodextrin at 25 degrees C suggested that the native structures of the three proteins are essentially the same. Differential scanning calorimetry of the thermal unfolding of the proteins showed that the unfolding temperature t(1/2) of the two mutant proteins decreased by about 10 degrees C as compared to the wild-type protein at pH 7.0. At t(1/2) of the wild-type protein (52.7 degrees C), the mutant proteins destabilized by about 10 kJ mol(-1) in terms of the Gibbs energy change. It was found that the mutant proteins were quite stabilized in terms of enthalpy, but that a higher entropy change overwhelmed the enthalpic effect, resulting in destabilization.

    DOI: 10.1271/bbb.70098

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  • Differential Scanning Calorimetry of the Effects of Ca^<2+> on the Thermal Unfolding of Pseudomonas cepacia Lipase Reviewed

    TANAKA Akiyoshi, SUGIMOTO Hayuki, MUTA Yuko, MIZUNO Takafumi, SENOO Keishi, OBATA Hitoshi, INOUYE Kuniyo

    Bioscience, Biotechnology, and Biochemistry   67 ( 1 )   207 - 210   2003.1

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    &nbsp;&nbsp;Thermal unfolding of <i>P. cepacia</i> lipase was observed by adiabatic differential scanning microcalorimetry in the absence and presence of calcium ions at pH 8, and thermodynamic parameters of unfolding were evaluated to analyze the unfolding mechanism of the enzyme. The temperature of unfolding was higher at higher concentrations of Ca<sup>2+</sup>. From the Ca<sup>2+</sup> concentration-dependence of the unfolding temperature, the number of calcium ions that dissociated from the enzyme molecule upon unfolding was estimated to be one. These results confirmed the validity of the unfolding mechanism proposed previously: NCa<sup>2+</sup>↔D+Ca<sup>2+</sup>, where N and D represent the native and denatured states, respectively, of the enzyme.<br>

    DOI: 10.1271/bbb.67.207

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  • Regulation of the chitin degradation and utilization system by the ChiX small RNA in Serratia marcescens 2170 Reviewed

    Kazushi Suzuki, Mari Shimizu, Naomi Sasaki, Chisana Ogawa, Haruka Minami, Hayuki Sugimoto, Takeshi Watanabe

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   80 ( 2 )   376 - 385   2016.2

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    Serratia marcescens 2170 produces three different types of chitinases and chitin-binding protein CBP21. We found that transposon insertion into the 5 untranslated region (5 UTR) of chiPQ-ctb led to defective chitinase and CBP21 production. ChiX small RNA possessed the complementary sequence of the 5 UTRs of the chiPQ-ctb and chiR and repressed the expression of chiP and chiR. ChiX was detected in a medium containing glucose, glycerol, GlcNAc, and (GlcNAc)(2), but the expression of both chiP and chiR was only observed in a medium containing (GlcNAc)(2). chiX mutant produced chitinases, CBP21, and chitobiase without induction. chiP transcripts were more abundant than those of chiR or chiX in a medium containing (GlcNAc)(2). These results suggest that the constitutively expressed ChiX binds to the highly abundant chiP 5 UTR, thereby leading to the induction of chiR mRNA translation and the subsequent expression of chitinases and CBP21.

    DOI: 10.1080/09168451.2015.1083399

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  • Construction of a CBP2l-defective mutant of Serratia marcescens 2170 Reviewed

    SUGIMOTO Hayuki

    21   46 - 51   2015

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    SUGIMOTO Hayuki

    応用糖質科学   4   107 - 112   2014

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  • Construction and basic characterization of deletion mutants of the genes involved in chitin utilization by Serratia marcescens 2170 Reviewed

    Shinya Takanao, Syouta Honma, Takuma Miura, Chisana Ogawa, Hayuki Sugimoto, Kazushi Suzuki, Takeshi Watanabe

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   78 ( 3 )   524 - 532   2014

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    In order to elucidate the roles of ChiP, ChiQ, and ChiX in chitin utilization by Serratia marcescens 2170, the construction of single-gene deletion mutants of the chiP, chiQ, and chiX genes was attempted by allelic exchange mutagenesis. Delta chiP formed smaller clearing zones and Delta chiX formed larger ones than wild-type 2170 on an agar plate containing colloidal chitin. Delta chiP grew slowly on the lower concentration of (GlcNAc)(2), and there was essentially no growth on chitin oligosaccharides larger than (GlcNAc)(3). The gene product of chiP was detected in the outer membrane fraction, consistently with the hypothesis that chiP encodes outer membrane chitoporin. Deletion of chiQ decreased and that of chiX increased the growth rates on chitin oligosaccharides. These observations strongly suggest that all three genes are involved in chitin utilization and that the deletion mutants obtained in this study might prove useful tools to clarify the details of the chitin utilization system of this bacterium.

    DOI: 10.1080/09168451.2014.882755

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  • Identification of a Csr system in Serratia marcescens 2170 Reviewed

    Manabu Ito, Kazuki Nomura, Hayuki Sugimoto, Takeshi Watanabe, Kazushi Suzuki

    JOURNAL OF GENERAL AND APPLIED MICROBIOLOGY   60 ( 2 )   79 - 88   2014

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    The carbon storage regulator (Csr) global regulatory system is conserved in many eubacteria and coordinates the expression of various genes that facilitate adaptation during the major physiological growth phase. The Csr system in Escherichia coli comprises an RNA-binding protein, CsrA; small non-coding RNAs, CsrB and CsrC; and a decay factor for small RNAs, CsrD. In this study, we identified the Csr system in Serratia marcescens 2170. S. marcescens CsrA was 97% identical to E. coli CsrA. CsrB and CsrC RNAs had typical stem-loop structures, including a GGA motif that is the CsrA binding site. CsrD was composed of N-terminal two times transmembrane region and HAMP-like, GGDEF, and EAL domains. Overexpression of S. marcescens csr genes complemented the phenotype of E. coli csr mutants. S. marcescens CsrD affected the decay of CsrB and CsrC RNAs in E. coli. These results suggest that the Csr system in S. marcescens is composed of an RNA-binding protein, two Csr small RNAs, and a decay factor for Csr small RNAs.

    DOI: 10.2323/jgam.60.79

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  • Involvement of Gln679, in addition to Trp687, in chitin-binding activity of the chitin-binding domain of chitinase A1 from Bacillus circulans WL-12 Reviewed

    Masashi Hara, Hayuki Sugimoto, Michio Uemura, Ken-ichi Akagi, Kazushi Suzuki, Takahisa Ikegami, Takeshi Watanabe

    JOURNAL OF BIOCHEMISTRY   154 ( 2 )   185 - 193   2013.8

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    Chitinase A1 (ChiA1) from Bacillus circulans WL-12 comprises an N-terminal catalytic domain, two fibronectin type III domains, and a C-terminal chitin-binding domain (ChBD). The ChBD of ChiA1 (ChBD(ChiA1)) belongs to carbohydrate-binding module (CBM) family 12 and specifically binds to insoluble or crystalline chitin. It has been suggested that tryptophan-687 (Trp687) is involved in the chitin-binding activity of this ChBD. Site-directed mutagenesis was used to identify additional amino acid residues required for chitin-binding activity of this domain. Furthermore, a total of 14 amino acid residues in ChBD(ChiA1) were carefully selected, and it was found that mutation of Gln679, which is not well-conserved in CBM family 12, significantly decreased the binding activity to colloidal chitin. A nuclear magnetic resonance study demonstrated that neither the Q679A nor the W687A mutation altered the overall structure of ChBD(ChiA1). Therefore, Gln679 was identified as a new residue that is involved in the chitin-binding activity of ChBD(ChiA1) in addition to Trp687. However, the mechanism of chitin binding by ChBD is still unknown.

    DOI: 10.1093/jb/mvt043

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  • Regulation of Chitinase Production by the 5 '-Untranslated Region of the ybfM in Serratia marcescens 2170 Reviewed

    Tadayuki Toratani, Kazushi Suzuki, Mari Shimizu, Hayuki Sugimoto, Takeshi Watanabe

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   76 ( 10 )   1920 - 1924   2012.10

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    Serratia marcescens 2170 produces three chitinases and the chitin-binding protein CBP21, and efficiently degrades insoluble and crystalline chitin. The three chitinases and CBP21 are induced by N,N'-diacetylchitobiose [(GlcNAc)(2)], the major product of chitin hydrolysis by S. marcescens chitinases. We have found that uptake of both (GlcNAc)(2) and N-acetylglucosamine (GlcNAc) is important for the efficient utilization of (GlcNAc)(2) because (GlcNAc)(2) is less efficiently fermented by S. marcescens 2170 in the absence of chitobiase. In order to determine the mechanism of utilization of the degradation products of chitin by S. marcescens, chitobiase deficient transposon mutants were screened. A transposon present in chitobiase-deficient mutants was inserted into the ybfMN-ctb cluster. The mutants produced chitinases, except for TT327, in which a transposon was inserted into the 5'-untranslated region (5'-UTR) of ybfM. Ectopic expression of this region in TT327 restored chitinase production. These results indicate that the 5'-UTR of ybfM is important for chitinase induction in S. marcescens.

    DOI: 10.1271/bbb.120403

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  • Phosphocholine-Containing Glycosyl Inositol-Phosphoceramides from Trichoderma viride Induce Defense Responses in Cultured Rice Cells Reviewed

    Ryosuke Uchiyama, Kazuhiro Aoki, Hayuki Sugimoto, Nobuko Taka, Takane Katayama, Saki Itonori, Mutsumi Sugita, Fang-Sik Che, Hidehiko Kumagai, Kenji Yamamoto

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   73 ( 1 )   74 - 78   2009.1

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    We isolated two major zwitterionic glycosphingolipids (ZGLs) from the phytopathogenic filamentous fungus Trichoderma viride. Structural analyses showed that the ZGLs (designated Tv-ZGL2 and Tv-ZGL3) were the same as the glycosphingolipids ZGL2 and ZGL4 from Acremonium sp., which are described in our previous paper. ZGLs have the following structure: Man(alpha 1-6)GlcN(alpha 1-2)Ins-P-Cer (Tv-ZGL2) and phosphocholine (PC) -&gt; 6Man(alpha 1-6)GlcN(alpha 1-2)Ins-P-Cer (Tv-ZGL3). To determine whether these ZGLs have functional roles in plant-fungus interaction, we tested to determine whether they would induce defense responses in cultured rice cells. We found that T. viride's ZGLs elicited expression of the PAL and PBZ1 genes, both of which are associated with pathogen resistance. Tv-ZGL2 induced cell death at a moderate rate. Tv-ZGL3, which contains a PC moiety, induced a high level of cell death in rice cells.

    DOI: 10.1271/bbb.80480

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  • Structural and thermodynamic analyses of solute-binding protein from Bifidobacterium longum specific for core 1 disaccharide and lacto-N-biose I Reviewed

    Ryuichiro Suzuki, Jun Wada, Takane Katayama, Shinya Fushinobu, Takayoshi Wakagi, Hirofumi Shoun, Hayuki Sugimoto, Akiyoshi Tanaka, Hidehiko Kumagai, Hisashi Ashida, Motomitsu Kitaoka, Kenji Yamamoto

    JOURNAL OF BIOLOGICAL CHEMISTRY   283 ( 19 )   13165 - 13173   2008.5

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Recently, a gene cluster involving a phosphorylase specific for lacto-N-biose I (LNB; Gal beta 1-3GlcNAc) and galacto-N-biose (GNB; Gal beta 1-3GalNAc) has been found in Bifidobacterium longum. We showed that the solute-binding protein of a putative ATP-binding cassette-type transporter encoded in the cluster crystallizes only in the presence of LNB or GNB, and therefore we named it GNB/LNB-binding protein (GL-BP). Isothermal titration calorimetry measurements revealed that GL-BP specifically binds LNB and GNB with K-d values of 0.087 and 0.010 mu M, respectively, and the binding process is enthalpy-driven. The crystal structures of GL-BP complexed with LNB, GNB, and lacto-N-tetraose ( Gal beta 1-3GlcNAc beta 1-3Gal beta 1-4Glc) were determined. The interactions between GL-BP and the disaccharide ligands mainly occurred through water-mediated hydrogen bonds. In comparison with the LNB complex, one additional hydrogen bond was found in the GNB complex. These structural characteristics of ligand binding are in agreement with the thermodynamic properties. The overall structure of GL-BP was similar to that of maltose-binding protein; however, the mode of ligand binding and the thermodynamic properties of these proteins were significantly different.

    DOI: 10.1074/jbc.M709777200

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  • 2P-079 Influences of the disulfide bond of the starch-binding domain of glucoamylase on the protein stability and refolding(The 46th Annual Meeting of the Biophysical Society of Japan)

    Sugimoto Hayuki, Nakaura Miho, Nishimura Shigenori, Karita Shuichi, Miyake Hideo, Tanaka Akiyoshi

    Seibutsu Butsuri   48   S87   2008

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    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.48.S87_3

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Books

  • 化学便覧 基礎編 改訂6版

    日本化学会( Role: Contributor ,  10章「熱的性質」 6.4 高分子)

    丸善出版  2021.1  ( ISBN:9784621305218

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    Total pages:xviii, 1509p   Language:Japanese

    CiNii Books

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  • 熱量測定・熱分析ハンドブック

    日本熱測定学会( Role: Contributor ,  5章 4.31 熱量測定とNMR)

    丸善  2020.8  ( ISBN:9784621305072

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    Total pages:xxiii, 363p   Language:Japanese

    CiNii Books

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MISC

  • Regulation of chitinase gene expression by incorporation of chitin degradation products and small RNA in Serratia marcescens

    SUZUKI Kazushi, SASAKI Naomi, OGAWA Chisana, TAKANO Shinya, SUGIMOTO Hayuki, WATANABE Takeshi

    18 ( 2 )   132 - 133   2012.7

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Research Projects

  • Hyperactivation of redox enzymes in confined nano-environments

    Grant number:18H01719

    2018.4 - 2021.3

    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

    Tsujimura Seiya

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    Grant amount:\17550000 ( Direct Cost: \13500000 、 Indirect Cost:\4050000 )

    We demonstrated the hyperactivation of redox enzymes and the improvement of their durability by utilizing a meso-reaction field (=electrode) with a scale of several tens of nanometers. We elucidated the complex interaction between microspace, enzymes, water, and ions, and we clarified the effects of mesostructure and environmental factors in the space on the enzymes. In this way, we have provided a novel design for controlling nanospace and solution environment around the enzyme beyond the conventional morphology control. Furthermore, we developed porous electrode materials suitable for new enzymatic electrode reactions, leading a dramatic improvement in the performance of enzyme-based bioelectronics devices.

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  • Elucidation of the mechanism underlying processive hydrolysis of crystalline chitin by bacterial chitinases.

    Grant number:15K07355

    2015.4 - 2018.3

    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

    Watanabe Takeshi, IGARASHI Kiyohiko, SUZUKI Kazushi, SUGIMOTO Hayuki

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    Grant amount:\4940000 ( Direct Cost: \3800000 、 Indirect Cost:\1140000 )

    The ultimate goal of this research is proposition of the universal and evolved model of the mechanism for crystalline chitin degradation and utilization by chitinolytic bacteria. Therefore, "the role of the protein (CBP21) that promote degradation of crystalline chitin", "the importance of CBP21 for the biological and ecological function of the chitinolytic bacteria", "the mechanism for biodegradation of α-chitin that exists in nature more abundantly" were studied. As a results, it became clear that CBP21 is especially important for degradation and utilization of chitin close to natural condition, an amino acid residue with aromatic ring of big resonating structure at the entrance of the catalytic cleft of chitinase is very important for hydrolysis of highly crystalline chitin substrate, and that one chitinase molecule jump to another chitin chain in different orientation and begins hydrolysis in opposite direction.

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  • Elucidation of the role of aromatic amino acid residues in crystalline chitin hydrolysis and development to the mechanism of a-chitin hydrolysis

    Grant number:24580104

    2012.4 - 2015.3

    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

    WATANABE TAKESHI, IGARASHI Kiyohiko, SUZUKI Kazushi, SUGIMOTO Hayuki

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    Grant amount:\5460000 ( Direct Cost: \4200000 、 Indirect Cost:\1260000 )

    Chitin is a crystalline and rigid structural polysaccharide present in a variety of organisms. Elucidation of the mechanism underlying in the enzymatic degradation of crystalline chitin is very important not only from enzymological and biological view points, but also for utilization of biomass resources. In this study, critical roles of the aromatic amino acid residues, which are present on the surface of chitinase molecule and inside of its catalytic cleft, in processive hydrolysis of crystalline chitin was demonstrated.

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  • Elucidation of the denatured structure of protein in equilibrium with the native one under a physiological condition.

    Grant number:21570173

    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

    SEGAWA Shin-ichi, NODA Yasuo, YUTANI Katsuhide, SUGIMOTO Hayuki

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

    We have studied partially ordered structures remaining in unstructured disulfide-deficient variants of lysozyme by means of NMR and H/D exchange methods. In this residual structure, two residues of I55 and L56 involved in the bottom of the α-sheet were located in a hydrophobic pocket formed with A, B and C-helices in the β-domain. Lysozyme folding appears to be initiated around this site. On the other hand, H/D exchange reactions were studied in the native state of pyrrolidone carboxyl peptidase(PCP). The native structure was found to be divided into two domains by structural changes near at the residue 70.

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  • 基質をスライドさせて連続反応を行う結晶性多糖分解酵素・キチナーゼの作動機構の 解明

    杉本華幸

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    Authorship:Principal investigator  Grant type:Competitive

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Teaching Experience (researchmap)

  • 応用微生物学

    Institution name:新潟大学農学部

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  • 遺伝子工学

    Institution name:新潟大学農学部

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Teaching Experience

  • スタディ・スキルズAIb

    2023
    Institution name:新潟大学

  • 統合化学入門

    2023
    Institution name:新潟大学

  • スタディ・スキルズAIIb

    2023
    Institution name:新潟大学

  • Topics in Biotechnology and Biochemistry

    2022
    Institution name:新潟大学

  • 応用生命科学セミナー

    2022
    Institution name:新潟大学

  • 微生物の物理化学

    2022
    Institution name:新潟大学

  • 醸造学

    2021
    Institution name:新潟大学

  • スタディ・スキルズAIc

    2021
    Institution name:新潟大学

  • スタディ・スキルズAIIc

    2021
    Institution name:新潟大学

  • 応用生命科学実験

    2020
    Institution name:新潟大学

  • 卒業論文Ⅰ

    2020
    Institution name:新潟大学

  • 応用生命科学演習Ⅰ

    2020
    Institution name:新潟大学

  • 科学英語演習

    2020
    Institution name:新潟大学

  • 応用生命科学演習Ⅱ

    2020
    Institution name:新潟大学

  • 卒業論文Ⅱ

    2020
    Institution name:新潟大学

  • Topics in Biotechnology and Biochemistry

    2020
    -
    2022
    Institution name:新潟大学

  • 農学入門Ⅱ

    2020
    -
    2021
    Institution name:新潟大学

  • 農学入門Ⅰ

    2020
    -
    2021
    Institution name:新潟大学

  • 食品科学概論

    2020
    -
    2021
    Institution name:新潟大学

  • 応用生命・食品科学特論

    2020
    Institution name:新潟大学

  • 分子微生物学特論

    2020
    Institution name:新潟大学

  • 応用生命・食品科学セミナーⅠ

    2020
    Institution name:新潟大学

  • 日本酒学A-2

    2019
    -
    2020
    Institution name:新潟大学

  • 日本酒学A-1

    2019
    -
    2020
    Institution name:新潟大学

  • 生命を知る

    2018
    Institution name:新潟大学

  • 応用微生物学

    2017
    Institution name:新潟大学

  • 微生物機能学

    2017
    Institution name:新潟大学

  • 基礎化学

    2017
    Institution name:新潟大学

  • 応用生命・食品科学概論

    2017
    -
    2020
    Institution name:新潟大学

  • 遺伝子工学

    2016
    Institution name:新潟大学

  • 醸造学

    2016
    Institution name:新潟大学

  • 微生物学実験

    2014
    Institution name:新潟大学

  • 分子生命科学演習Ⅰ

    2014
    -
    2019
    Institution name:新潟大学

  • 分子生命科学演習Ⅱ

    2014
    -
    2018
    Institution name:新潟大学

  • 分子生命科学実験

    2014
    -
    2018
    Institution name:新潟大学

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