2024/06/14 更新

写真a

タキハラ ハヤト
瀧原 速仁
TAKIHARA Hayato
所属
教育研究院 医歯学系 特任助教
医歯学総合研究科 特任助教
職名
特任助教
外部リンク

学位

  • 博士 (生物資源科学) ( 2015年3月   日本大学 )

研究キーワード

  • 微生物生態学

  • バイオインフォマティクス

  • メタゲノム

  • 環境微生物

  • 糖鎖関連遺伝子

研究分野

  • ライフサイエンス / システムゲノム科学  / バイオインフォマティクス

  • ライフサイエンス / 応用微生物学  / メタゲノム

経歴(researchmap)

  • 新潟大学大学院 医歯学総合研究科   バイオインフォーマティクス分野   特任助教

    2020年11月 - 現在

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    国名:日本国

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  • 新潟大学   大学院医歯学総合研究科   博士研究員

    2015年7月 - 2020年10月

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経歴

  • 新潟大学   医歯学総合研究科   特任助教

    2020年11月 - 現在

学歴

  • 日本大学   大学院 生物資源科学研究科   応用生命科学専攻

    2009年4月 - 2011年3月

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  • 日本大学   生物資源科学部   応用生物科学科

    2005年4月 - 2009年3月

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論文

  • Alkane-Translocated Cells of <i>Rhodococcus</i> Strains Utilize Dissolved Oxygen in the Alkane Phase of an Aqueous-Alkane Two-phase Culture

    Noriyuki Iwabuchi, Hayato Takihara

    Bioscience, Biotechnology, and Biochemistry   2024年5月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Oxford University Press (OUP)  

    Abstract

    To clarify the growth mechanisms of Rhodococcus in the alkane phase, we measured oxygen utilization in the alkane phase. The results showed that dissolved oxygen decreased significantly when viable cells were present in the alkane phase. The findings suggested that Rhodococcus strains can grow in alkanes and utilize the resident dissolved oxygen.

    DOI: 10.1093/bbb/zbae074

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  • Genetic engineering employing MPB70 and its promoter enables efficient secretion and expression of foreign antigen in bacillus Calmette Guérin (BCG) Tokyo. 国際誌

    Atsuki Takeishi, Amina K Shaban, Taichi Kakihana, Hayato Takihara, Shujiro Okuda, Hidekazu Osada, Desak Nyoman Surya Suameitria Dewi, Yuriko Ozeki, Yutaka Yoshida, Akihito Nishiyama, Yoshitaka Tateishi, Yuki Aizu, Yasushi Chuma, Kazuyo Onishi, Daisuke Hayashi, Saburo Yamamoto, Tetsu Mukai, Manabu Ato, Duong Huu Thai, Huynh Thi Thao Nhi, Tsuyoshi Shirai, Satoshi Shibata, Fumiko Obata, Jun Fujii, Seiya Yamayoshi, Maki Kiso, Sohkichi Matsumoto

    Microbiology and immunology   2024年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Vaccination is an important factor in public health. The recombinant bacillus Calmette Guérin (rBCG) vaccine, which expresses foreign antigens, is expected to be a superior vaccine against infectious diseases. Here, we report a new recombination platform in which the BCG Tokyo strain is transformed with nucleotide sequences encoding foreign protein fused with the MPB70 immunogenic protein precursor. By RNA-sequencing, mpb70 was found to be the most transcribed among all known genes of BCG Tokyo. Small oligopeptide, namely, polyhistidine tag, was able to be expressed in and secreted from rBCG through a process in which polyhistidine tag fused with intact MPB70 were transcribed by an mpb70 promoter. This methodology was applied to develop an rBCG expressing the receptor binding domain (RBD) of severe acute respiratory syndrome coronavirus 2. Immunoblotting images and mass spectrometry data showed that RBD was also secreted from rBCG. Sera from mice vaccinated with the rBCG showed a tendency of weak neutralizing capacity. The secretion was retained even after a freeze-drying process. The freeze-dried rBCG was administered to and recovered from mice. Recovered rBCG kept secreting RBD. Collectively, our recombination platform offers stable secretion of foreign antigens and can be applied to the development of practical rBCGs.

    DOI: 10.1111/1348-0421.13116

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  • Mycobacterial DNA-binding protein 1 is critical for BCG survival in stressful environments and simultaneously regulates gene expression. 国際誌

    Amina K Shaban, Gebremichal Gebretsadik, Mariko Hakamata, Hayato Takihara, Erina Inouchi, Akihito Nishiyama, Yuriko Ozeki, Yoshitaka Tateishi, Yukiko Nishiuchi, Takehiro Yamaguchi, Naoya Ohara, Shujiro Okuda, Sohkichi Matsumoto

    Scientific reports   13 ( 1 )   14157 - 14157   2023年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Survival of the live attenuated Bacillus Calmette-Guérin (BCG) vaccine amidst harsh host environments is key for BCG effectiveness as it allows continuous immune response induction and protection against tuberculosis. Mycobacterial DNA binding protein 1 (MDP1), a nucleoid associated protein, is essential in BCG. However, there is limited knowledge on the extent of MDP1 gene regulation and how this influences BCG survival. Here, we demonstrate that MDP1 conditional knockdown (cKD) BCG grows slower than vector control in vitro, and dies faster upon exposure to antibiotics (bedaquiline) and oxidative stress (H2O2 and menadione). MDP1-cKD BCG also exhibited low infectivity and survival in THP-1 macrophages and mice indicating possible susceptibility to host mediated stress. Consequently, low in vivo survival resulted in reduced cytokine (IFN-gamma and TNF-alpha) production by splenocytes. Temporal transcriptome profiling showed more upregulated (81-240) than downregulated (5-175) genes in response to MDP1 suppression. Pathway analysis showed suppression of biosynthetic pathways that coincide with low in vitro growth. Notable was the deferential expression of genes involved in stress response (sigI), maintenance of DNA integrity (mutT1), REDOX balance (WhiB3), and host interactions (PE/PE_PGRS). Thus, this study shows MDP1's importance in BCG survival and highlights MDP1-dependent gene regulation suggesting its role in growth and stress adaptation.

    DOI: 10.1038/s41598-023-40941-9

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  • Glycan-related genes in human gut microbiota exhibit differential distribution and diversity in carbohydrate degradation and glycan synthesis. 国際誌

    Hayato Takihara, Shujiro Okuda

    Frontiers in molecular biosciences   10   1137303 - 1137303   2023年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Interactions between humans and the gut microbiome occur by supplying nutrients to gut epithelial cells via short-chain fatty acids obtained from dietary carbohydrates or mucins and activating immunity via mucins' degradation. The degradation of carbohydrates derived from food is an important function for organisms to obtain energy. However, since humans possess only 17 genes encoding carbohydrate-degrading enzymes, the gut microbiome is responsible for degrading plant-derived polysaccharides. Using the method for extracting glycan-related genes from the metagenomes constructed thus far, we calculated the distribution and abundance of different glycan-related genes in the healthy human gut metagenome. Glycan-related genes showed an abundance of 0.64-11.00, indicating large individual differences. However, the distribution of the classes of glycan-related genes was similar between the samples. In addition, the function of carbohydrate degradation was divided into three clusters, showing high diversity; however, the synthesis function was not divided, indicating low diversity. The substrates of enzymes for carbohydrate degradation between clusters were either plant-derived polysaccharides or biased toward degrading polysaccharides derived from other sources. These functional biases differ depending on the type of microorganism used. Based on these findings, we predicted that 1) diversity will be constant because the influence on the host by the transferase of gut bacteria is a function derived from the genome, and 2) diversity will be high because the influence on the host by the hydrolase of gut bacteria is affected by incoming dietary carbohydrates.

    DOI: 10.3389/fmolb.2023.1137303

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  • Differential analysis of microbiomes in mucus and tissues obtained from colorectal cancer patients. 国際誌

    Yosuke Tajima, Shujiro Okuda, Tsunekazu Hanai, Junichiro Hiro, Koji Masumori, Yoshikazu Koide, Tadahiro Kamiya, Yeongcheol Cheong, Gaku Inaguma, Yoshifumi Shimada, Toshifumi Wakai, Hayato Takihara, Shingo Akimoto, Hiroshi Matsuoka, Ichiro Uyama, Koichi Suda

    Scientific reports   12 ( 1 )   18193 - 18193   2022年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The outer mucus layer of the colorectal epithelium is easily removable and colonized by commensal microbiota, while the inner mucus layer is firmly attached to the epithelium and devoid of bacteria. Although the specific bacteria penetrating the inner mucus layer can contact epithelial cells and trigger cancer development, most studies ignore the degree of mucus adhesion at sampling. Therefore, we evaluated whether bacteria adhering to tissues could be identified by removing the outer mucus layer. Our 16S rRNA gene sequencing analysis of 18 surgical specimens of human colorectal cancer revealed that Sutterella (P = 0.045) and Enterobacteriaceae (P = 0.045) were significantly enriched in the mucus covering the mucosa relative to the mucosa. Rikenellaceae (P = 0.026) was significantly enriched in the mucus covering cancer tissues compared with those same cancer tissues. Ruminococcaceae (P = 0.015), Enterobacteriaceae (P = 0.030), and Erysipelotrichaceae (P = 0.028) were significantly enriched in the mucus covering the mucosa compared with the mucus covering cancers. Fusobacterium (P = 0.038) was significantly enriched in the mucus covering cancers compared with the mucus covering the mucosa. Comparing the microbiomes of mucus and tissues with mucus removed may facilitate identifying bacteria that genuinely invade tissues and affect tumorigenesis.

    DOI: 10.1038/s41598-022-21928-4

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  • Unveiling microbiome profiles in human inner body fluids and tumor tissues with pancreatic or biliary tract cancer. 国際誌

    Shujiro Okuda, Yuki Hirose, Hayato Takihara, Akiko Okuda, Yiwei Ling, Yosuke Tajima, Yoshifumi Shimada, Hiroshi Ichikawa, Kazuyasu Takizawa, Jun Sakata, Toshifumi Wakai

    Scientific reports   12 ( 1 )   8766 - 8766   2022年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    With the discovery of bacterial symbiosis in the tissues of various cancers, the study of the tumor microbiome is attracting a great deal of attention. Anatomically, since the gastrointestinal tract, liver, and pancreas form a continuous ductal structure, the microbiomes in the digestive juices of these organs may influence each other. Here, we report a series of microbiome data in tumor-associated tissues such as tumor, non-tumor, and lymph nodes, and body fluids such as saliva, gastric juice, pancreatic juice, bile, and feces of patients with pancreatic or biliary tract cancers. The results show that the microbiome of tumor-associated tissues has a very similar bacterial composition, but that in body fluids has different bacterial composition which varies by location, where some bacteria localize to specific body fluids. Surprisingly, Akkermansia was only detected in the bile of patients with biliary tract cancer and its presence was significantly associated with the performance of external biliary drainage (P = 0.041). Furthermore, we found that tumor-associated tissues and body fluids in deep inner body are mostly inhabited by unidentified and uncharacterized bacteria, suggesting that such bacteria may be potential targets for precision therapy in the future.

    DOI: 10.1038/s41598-022-12658-8

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  • Monitoring IgG against Mycobacterium tuberculosis proteins in an Asian elephant cured of tuberculosis that developed from long-term latency. 国際誌

    Satoshi Ishikawa, Yuriko Ozeki, Satomi Suga, Yasuhiko Mukai, Haruka Kobayashi, Erina Inouchi, Shaban A Kaboso, Gebremichal Gebretsadik, Desak Nyoman Surya Suameitria Dewi, Akihito Nishiyama, Yoshitaka Tateishi, Hayato Takihara, Shujiro Okuda, Shiomi Yoshida, Naoaki Misawa, Sohkichi Matsumoto

    Scientific reports   12 ( 1 )   4310 - 4310   2022年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Tuberculosis (TB) is fatal in elephants, hence protecting elephants from TB is key not only in the conservation of this endangered animal, but also to prevent TB transmission from elephants to humans. Most human TB cases arise from long-term asymptomatic infections. Significant diagnostic challenges remain in the detection of both infection and disease development from latency in elephants due to their huge bodies. In this study, we assessed cryopreserved sera collected for over 16 years, from the first Japanese treatment case of elephant TB. Semi-quantification of IgG levels to 11 proteins showed high detection levels of 3 proteins, namely ESAT6/CFP10, MPB83 and Ag85B. The level of IgG specific to these 3 antigens was measured longitudinally, revealing high and stable ESAT6/CFP10 IgG levels regardless of onset or treatment. Ag85B-specifc IgG levels were largely responsive to onset or treatment, while those of MPB83 showed intermediate responses. These results suggest that ESAT6/CFP10 is immunodominant in both asymptomatic and symptomatic phases, making it useful in the detection of infection. On the other hand, Ag85B has the potential to be a marker for the prediction of disease onset and in the evaluation of treatment effectiveness in elephants.

    DOI: 10.1038/s41598-022-08228-7

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  • Functional glyco-metagenomics elucidates the role of glycan-related genes in environments. 国際誌

    Hayato Takihara, Nobuaki Miura, Kiyoko F Aoki-Kinoshita, Shujiro Okuda

    BMC bioinformatics   22 ( 1 )   505 - 505   2021年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    BACKGROUND: Glycan-related genes play a fundamental role in various processes for energy acquisition and homeostasis maintenance while adapting to the environment in which the organism exists; however, their role in the microbiome in the environment is unclear. METHODS: Sequence alignment was performed between known glycan-related genes and complete genomes of microorganisms, and optimal parameters for identifying glycan-related genes were determined based on the alignments. Using the constructed scheme (> 90% of identity and > 25 aa of alignment length), glycan-related genes in various environments were identified from 198 different metagenome data. RESULTS: As a result, we identified 86.73 million glycan-related genes from the metagenome data. Among the 12 environments classified in this study, the percentage of glycan-related genes was high in the human-associated environment, suggesting that these environments utilize glycan metabolism better than other environments. On the other hand, the relative abundances of both glycoside hydrolases and glycosyltransferases surprisingly had a coverage of over 80% in all the environments. These glycoside hydrolases and glycosyltransferases were classified into two groups of (1) general enzyme families identified in various environments and (2) specific enzymes found only in certain environments. The general enzyme families were mostly from genes involved in monosaccharide metabolism, and most of the specific enzymes were polysaccharide degrading enzymes. CONCLUSION: These findings suggest that environmental microorganisms could change the composition of their glycan-related genes to adapt the processes involved in acquiring energy from glycans in their environments. Our functional glyco-metagenomics approach has made it possible to clarify the relationship between the environment and genes from the perspective of carbohydrates, and the existence of glycan-related genes that exist specifically in the environment.

    DOI: 10.1186/s12859-021-04425-9

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  • Development of Novel PCR Assays for Improved Detection of Enterovirus D68. 国際誌

    Tatsuki Ikuse, Yuta Aizawa, Hayato Takihara, Shujiro Okuda, Kanako Watanabe, Akihiko Saitoh

    Journal of clinical microbiology   59 ( 11 )   JCM0115121   2021年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Enterovirus D68 (EV-D68) causes a range of clinical manifestations, including asthma-like illness, severe respiratory disease, and acute flaccid myelitis. EV-D68 has caused worldwide outbreaks since 2014 and is now recognized as a re-emerging infection in many countries. EV-D68-specific PCR assays are widely used for the diagnosis of EV-D68 infection; however, assay sensitivity is a concern because of genetic changes in recently circulated EV-D68. To address this, we summarized EV-D68 sequences from previously reported world outbreaks from 2014 through 2020 on GenBank, and found several mutations at the primer and probe binding sites of the existing EV-D68-specific PCR assays. Subsequently, we designed two novel assays corresponding to the recently reported EV-D68 sequences: an EV-D68-specific real-time and semi-nested PCR. In an analysis of 22 EV-D68-confirmed cases during a recent EV-D68 outbreak in Japan, the new real-time PCR had higher sensitivity than the existing assay (100% vs. 45%, P < 0.01) and a lower median Ct value (27.8 vs. 32.8, P = 0.005). Sensitivity was higher for the new non-nested PCR (91%) than for the existing semi-nested PCR assay (50%, P < 0.01). The specificity of the new real-time PCR was 100% using samples from non-EV-D68-infected cases (n = 135). In conclusion, our novel assays had higher sensitivity than the existing assay and might lead to more accurate diagnosis of recently circulating EV-D68. To prepare for future EV-D68 outbreaks, EV-D68-specific assays must be continuously monitored and updated.

    DOI: 10.1128/JCM.01151-21

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  • Extracellular DNA of slow growers of mycobacteria and its contribution to biofilm formation and drug tolerance. 国際誌

    Aleksandr Ilinov, Akihito Nishiyama, Hiroki Namba, Yukari Fukushima, Hayato Takihara, Chie Nakajima, Anna Savitskaya, Gebremichal Gebretsadik, Mariko Hakamata, Yuriko Ozeki, Yoshitaka Tateishi, Shujiro Okuda, Yasuhiko Suzuki, Yuri S Vinnik, Sohkichi Matsumoto

    Scientific reports   11 ( 1 )   10953 - 10953   2021年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DNA is basically an intracellular molecule that stores genetic information and carries instructions for growth and reproduction in all cellular organisms. However, in some bacteria, DNA has additional roles outside the cells as extracellular DNA (eDNA), which is an essential component of biofilm formation and hence antibiotic tolerance. Mycobacteria include life-threating human pathogens, most of which are slow growers. However, little is known about the nature of pathogenic mycobacteria's eDNA. Here we found that eDNA is present in slow-growing mycobacterial pathogens, such as Mycobacterium tuberculosis, M. intracellulare, and M. avium at exponential growth phase. In contrast, eDNA is little in all tested rapid-growing mycobacteria. The physiological impact of disrupted eDNA on slow-growing mycobacteria include reduced pellicle formation, floating biofilm, and enhanced susceptibility to isoniazid and amikacin. Isolation and sequencing of eDNA revealed that it is identical to the genomic DNA in M. tuberculosis and M. intracellulare. In contrast, accumulation of phage DNA in eDNA of M. avium, suggests that the DNA released differs among mycobacterial species. Our data show important functions of eDNA necessary for biofilm formation and drug tolerance in slow-growing mycobacteria.

    DOI: 10.1038/s41598-021-90156-z

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  • Higher genome mutation rates of Beijing lineage of Mycobacterium tuberculosis during human infection. 国際誌

    Mariko Hakamata, Hayato Takihara, Tomotada Iwamoto, Aki Tamaru, Atsushi Hashimoto, Takahiro Tanaka, Shaban A Kaboso, Gebremichal Gebretsadik, Aleksandr Ilinov, Akira Yokoyama, Yuriko Ozeki, Akihito Nishiyama, Yoshitaka Tateishi, Hiroshi Moro, Toshiaki Kikuchi, Shujiro Okuda, Sohkichi Matsumoto

    Scientific reports   10 ( 1 )   17997 - 17997   2020年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Mycobacterium tuberculosis (Mtb) strains of Beijing lineage have caused great concern because of their rapid emergence of drug resistance and worldwide spread. DNA mutation rates that reflect evolutional adaptation to host responses and the appearance of drug resistance have not been elucidated in human-infected Beijing strains. We tracked and obtained an original Mtb isolate of Beijing lineage from the 1999 tuberculosis outbreak in Japan, as well as five other isolates that spread in humans, and two isolates from the patient caused recurrence. Three isolates were from patients who developed TB within one year after infection (rapid-progressor, RP), and the other three isolates were from those who developed TB more than one year after infection (slow-progressor, SP). We sequenced genomes of these isolates and analyzed the propensity and rate of genomic mutations. Generation time versus mutation rate curves were significantly higher for RP. The ratio of oxidative versus non-oxidation damages induced mutations was higher in SP than RP, suggesting that persistent Mtb are exposed to oxidative stress in the latent state. Our data thus demonstrates that higher mutation rates of Mtb Beijing strains during human infection is likely to account for the higher adaptability and an emergence ratio of drug resistance.

    DOI: 10.1038/s41598-020-75028-2

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  • Innate Immune Responses in Serum and Cerebrospinal Fluid From Neonates and Infants Infected With Parechovirus-A3 or Enteroviruses. 国際誌

    Rie Habuka, Yuta Aizawa, Ryohei Izumita, Hisanori Domon, Yutaka Terao, Hayato Takihara, Shujiro Okuda, Akihiko Saitoh

    The Journal of infectious diseases   222 ( 4 )   681 - 689   2020年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    BACKGROUND: Parechovirus (PeV)-A3 and enteroviruses (EV) are the most common viruses causing sepsis and meningoencephalitis in neonates and young infants. Clinical manifestations of PeV-A3 infection are more severe than those of EV infection, and no pleocytosis with a positive polymerase chain reaction (PCR) result for PeV-A3 in cerebrospinal fluid (CSF) are characteristic findings. We hypothesized that innate immune responses to PeV-A3 and EV are distinct in serum and CSF. METHODS: We evaluated 22 cytokines/chemokines in serum and CSF from PeV-A3- or EV-infected patients younger than 4 months in Niigata, Japan, from 2015 through 2018. Infection was diagnosed with real-time PCR followed by sequencing. Febrile neonates and infants with sepsis-like syndrome who had negative bacterial culture and viral PCR for both PeV-A and EV were also included (non-PeV-A/EV patients). RESULTS: Among 192 febrile patients, we evaluated 16 PeV-A3-infected, 15 EV-infected, and 8 non-PeV-A/EV patients. Serum pro-/anti-inflammatory cytokine/chemokine levels were higher in PeV-A3-infected patients than in EV-infected patients (P < .02). Although most cytokine/chemokine were elevated in CSF from EV-infected patients, levels were low or undetectable in PeV-A3-infected and non-PeV-A/EV patients (P < .001). CONCLUSIONS: Distinct cytokine/chemokine patterns in serum and CSF may explain the different clinical manifestations of PeV-A3-infected and EV-infected neonates and young infants.

    DOI: 10.1093/infdis/jiaa131

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  • Rice Endosperm Protein Administration to Juvenile Mice Regulates Gut Microbiota and Suppresses the Development of High-Fat Diet-Induced Obesity and Related Disorders in Adulthood. 国際誌

    Yuki Higuchi, Michihiro Hosojima, Hideyuki Kabasawa, Shoji Kuwahara, Sawako Goto, Koji Toba, Ryohei Kaseda, Takahiro Tanaka, Nobutaka Kitamura, Hayato Takihara, Shujiro Okuda, Masayuki Taniguchi, Hitoshi Arao, Ichiei Narita, Akihiko Saito

    Nutrients   11 ( 12 )   2019年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Obesity and related disorders, which are increasing in adults worldwide, are closely linked to childhood diet and are associated with chronic inflammation. Rice endosperm protein (REP) intake during adulthood has been reported to improve lipid metabolism and suppress the progression of diabetic kidney disease in animal models. However, the effects of REP intake during childhood on adulthood health are unclear. Therefore, we used a mouse model to experimentally investigate the preconditioning effects of REP intake during childhood on the development of obesity and related disorders in adulthood. Male C57BL/6J mice were pair-fed a normal-fat diet containing casein or REP during the juvenile period and then a high-fat diet (HFD) containing casein or REP during adulthood. Mice fed REP during the juvenile period showed better body weight, blood pressure, serum lipid profiles, lipopolysaccharide (LPS)-binding protein levels, and glucose tolerance in adulthood than those fed casein during the juvenile period. HFD-induced renal tubulo-glomerular alterations and hepatic microvesicular steatosis were less evident in REP-fed mice than in casein-fed ones. REP intake during the juvenile period improved HFD-induced dysbiosis (i.e., Escherichia genus proliferation and reduced gut microbiota diversity), thereby suppressing endotoxin-related chronic inflammation. Indeed, REP-derived peptides showed antibacterial activity against Escherichia coli, a major producer of LPS. In conclusion, REP supplementation during the juvenile period may regulate the gut microbiota and thus suppress the development of obesity and related disorders in adulthood in mice.

    DOI: 10.3390/nu11122919

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  • Development of a Simple Nonbiological Method for Converting Lignin-Derived Aromatics into Nonaromatic Polymeric Substances with Fluorescent Activity

    Noriyuki Iwabuchi, Yuki Sakano, Hajime Takiguchi, Hayato Takihara, Michio Sunairi, Hiroshi Matsufuji

    ACS SUSTAINABLE CHEMISTRY & ENGINEERING   4 ( 8 )   4411 - 4416   2016年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    We recently reported that Pseudomonas sp. ITH-SA-1 can transform the lignin-derived aromatic compound syringaldehyde (SYAL) into nonaromatic polymeric substances with fluorescent activity (NAPSFA). NAPSFA are particularly rare organic substances that fluoresce despite the absence of aromatic rings. In this study, we developed a simple method for producing NAPSFA using a nonbiological process. Incubation of the SYAL metabolite 3-O-methyl gallate (3-MGA) produced by Pseudomonas sp. ITH-SA-1 with Marine Broth (MB) or Luria-Bertani broth (LB) produced fluorescence even in the absence of bacteria, suggesting that fluorescent substances were produced from 3-MGA by a nonbiological process. 3-MGA reacted with tryptone and peptone, which are the primary nitrogen and phosphate sources in MB and LB, to produce the fluorescent substances. The fluorescent substances produced by the reaction of 3-MGA with tryptone exhibited excitation and emission peaks at 370 and 505 nm, respectively, which are nearly identical to those of NAPSFA (365/498 nm). The average molecular weight of the fluorescent substances (4.2 kDa) was lower than that of NAPSFA (7.2 kDa). ATR-FTIR and NMR analyses revealed that the molecules contained no aromatic rings, similar to NAPSFA. Our results demonstrate that nonaromatic fluorescent substances can be synthesized via a simple chemical reaction.

    DOI: 10.1021/acssuschemeng.6b01009

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  • Mg(2+)-Dependent Control of the Spatial Arrangement of Rhodococcus erythropolis PR4 Cells in Aqueous-Alkane Two Phase Culture Containing n-Dodecane.

    Hayato Takihara, Yumiko Akase, Michio Sunairi, Noriyuki Iwabuchi

    Microbes and environments   31 ( 2 )   178 - 81   2016年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We recently reported that a close relationship exists between alkane carbon-chain length, cell growth, and translocation frequency in Rhodococcus. In the present study, we examined the regulation of the spatial arrangement of cells in aqueous-alkane two phase cultures. An analysis of the effects of minerals on cell localization revealed that changes in the concentration of MgSO4 in two phase cultures containing n-dodecane (C12) altered cell localization from translocation to adhesion and vice versa. Our results indicate that the spatial arrangement of cells in two phase culture systems is controlled through the regulation of MgSO4 concentrations.

    DOI: 10.1264/jsme2.ME15196

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  • Transformation of Lignin-Derived Aromatics into Nonaromatic Polymeric Substances with Fluorescent Activities (NAPSFA) by Pseudomonas sp ITH-SA-1

    Noriyuki Iwabuchi, Hajime Takiguchi, Takashi Hamaguchi, Hayato Takihara, Michio Sunairi, Hiroshi Matsufuji

    ACS Sustainable Chemistry & Engineering   3 ( 11 )   2678 - 2685   2015年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    We examined bacteria capable of transforming lignin-derived compounds into industrially or economically valuable substances from the seawater after the Great East Japan Earthquake-caused tsunami. Pseudomonas sp. ITH-SA-1 produced water-soluble fluorescent substances from the lignin-derived aromatic, syringaldehyde (SYAL). They: are polymeric substances derived from 3-O-methyl gallate produced through transformation of SYAL via syringate; are not known compounds reported previously; have excitation/emission peaks at 365/498 nm, respectively; and have an average molecular weight of 7.2 kDa. Despite the fact that aromatic structure generally plays an important role in the planarity and rigidity of organic fluorescent substance, the spectroscopic analyses revealed that aromatic rings were not detected in the molecules. Their activity is particularly rare and the biotransforming capabilities will contribute to the development of new basic techniques for the effective utilization of lignin.

    DOI: 10.1021/acssuschemeng.5b00503

    Web of Science

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  • Rhodococcus rhodochrous ATCC12674 becomes alkane-tolerant upon GroEL2 overexpression and survives in the n-octane phase in two phase culture.

    Hayato Takihara, Chiaki Matsuura, Jun Ogihara, Noriyuki Iwabuchi, Michio Sunairi

    Microbes and environments   29 ( 4 )   431 - 3   2014年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We recently reported that the overexpression of GroEL2 played an important role in increasing the alkane tolerance of Rhodococcus erythropolis PR4. In the present study, we examined the effects of the introduction of groEL2 on the alkane tolerance of other Rhodococcus strains. The introduction of groEL2 into Rhodococcus strains led to increased alkane tolerance. The translocation of R. rhodochrous ATCC12674 cells to and survival in the n-octane (C8) phase in two phase culture were significantly enhanced by the introduction of groEL2 derived from strain PR4, suggesting that engineering cells to overexpress GroEL2 represents an effective strategy for enhancing organic solvent tolerance in Rhodococcus.

    DOI: 10.1264/jsme2.ME14114

    PubMed

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  • Enhanced translocation and growth of Rhodococcus erythropolis PR4 in the alkane phase of aqueous-alkane two phase cultures were mediated by GroEL2 overexpression.

    Hayato Takihara, Jun Ogihara, Takao Yoshida, Shujiro Okuda, Mutsuyasu Nakajima, Noriyuki Iwabuchi, Michio Sunairi

    Microbes and environments   29 ( 4 )   346 - 52   2014年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We previously reported that R. erythropolis PR4 translocated from the aqueous to the alkane phase, and then grew in two phase cultures to which long-chain alkanes had been added. This was considered to be beneficial for bioremediation. In the present study, we investigated the proteins involved in the translocation of R. erythropolis PR4. The results of our proteogenomic analysis suggested that GroEL2 was upregulated more in cells that translocated inside of the pristane (C19) phase than in those located at the aqueous-alkane interface attached to the n-dodecane (C12) surface. PR4 (pK4-EL2-1) and PR4 (pK4-ΔEL2-1) strains were constructed to confirm the effects of the upregulation of GroEL2 in translocated cells. The expression of GroEL2 in PR4 (pK4-EL2-1) was 15.5-fold higher than that in PR4 (pK4-ΔEL2-1) in two phase cultures containing C12. The growth and cell surface lipophilicity of PR4 were enhanced by the introduction of pK4-EL2-1. These results suggested that the plasmid overexpression of groEL2 in PR4 (pK4-EL2-1) led to changes in cell localization, enhanced growth, and increased cell surface lipophilicity. Thus, we concluded that the overexpression of GroEL2 may play an important role in increasing the organic solvent tolerance of R. erythropolis PR4 in aqueous-alkane two phase cultures.

    DOI: 10.1264/jsme2.ME13158

    PubMed

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MISC

  • バイオインフォマティクスの世界(第15回) やってみようバイオインフォマティクス メタ16S解析編

    瀧原 速仁, 奥田 修二郎

    医学のあゆみ   282 ( 4 )   296 - 303   2022年7月

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    記述言語:日本語   出版者・発行元:医歯薬出版(株)  

    環境中の徴生物がどのような種で構成されているかを調べる解析法として、メタ16S解析が最もよく利用されている。この手法の強みとしてPCRプライマーによる16S rRNA遺伝子の増幅が利用てきること、リファレンス配列が十分にデータベース化されていること、微量のサンプルでも解析可能なことがあげられる。現在はハイスループットシーケンスを用いることによって簡便かつ高速に網羅解析することが可能となった。メタ16S解析に用いられるシーケンサーはIllumina社のHiSeqやMiSeqが利用され、1サンプルあたりのコストは2-3万円で可能てある。Illumina社のシーケンサーから得られた塩基配列のでータはFASTQファイルという形式で出力され、リード名、塩基配列、クオリティ値で構成される。本文章では、このFASTQファイルから始めて各微生物種の割リ当て、相対量の算出までのプロセスを説明する。(著者抄録)

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    その他リンク: https://search-tp.jamas.or.jp/index.php?module=Default&action=Link&pub_year=2022&ichushi_jid=J00060&link_issn=&doc_id=20220727010011&doc_link_id=%2Faa7ayuma%2F2022%2F028204%2F012%2F0296b0303%26dl%3D3&url=https%3A%2F%2Fwww.medicalonline.jp%2Fjamas.php%3FGoodsID%3D%2Faa7ayuma%2F2022%2F028204%2F012%2F0296b0303%26dl%3D3&type=MedicalOnline&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F00004_4.gif

  • 早期発症者と長期潜伏後発症者より分離した結核菌北京株のゲノム変異解析

    袴田 真理子, 瀧原 速仁, 岩本 朋忠, 田丸 亜貴, 尾関 百合子, 西山 晃史, 立石 善隆, 菊地 利明, 奥田 修二郎, 松本 壮吉

    結核   96 ( 3 )   83 - 86   2021年5月

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    記述言語:日本語   出版者・発行元:(一社)日本結核・非結核性抗酸菌症学会  

    [目的]全ゲノム解析を用いて,早期発症者と長期潜伏後発症者から分離した結核菌北京株のゲノム変異を解析した。[方法]1999年に中学校で発生した結核集団感染の接触者と二次感染者のうち2009年までに結核を発症した患者より分離した結核菌北京株6株と,別の事例で初発時と再発時の患者より分離した結核菌北京株2株の計8株の全ゲノム解析を行った。結核菌の突然変異率は,初発から1年以内に発症した早期発症者と1年以上の潜伏期を経てから発症または再燃した長期潜伏後発症者の2群間で比較した。[結果]結核菌北京株の突然変異率は,Lineage 4に属する結核菌よりも高く,結核菌北京株の高い病原性や薬剤耐性の要因である可能性が示唆された。遺伝子多型解析では,酸化的損傷に起因すると推定されている突然変異が長期潜伏後発症群のほうに多く,潜伏期間中にも薬剤耐性変異が起こる可能性が示唆された。[結論]結核菌北京株の高頻度の薬剤耐性化を防ぐためには,感染した結核菌の系統により治療法を検討する必要性も考えられた。今後,結核菌系統の特性を理解し,治療法を工夫することで結核の薬剤耐性化や重篤化を防ぐ有効な対策の構築につながることが期待される。(著者抄録)

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  • 世界結核デーにちなんで、世界の結核・抗酸菌症研究のこれまでと今 結核菌北京株のゲノム解析と組織透明化/3次元イメージング「CUBIC」による抗酸菌感染の生体内モニタリング

    袴田 真理子, 瀧原 速仁, 尾関 百合子, 西山 晃史, 立石 善隆, 大橋 璃子, 奥田 修二郎, 田井中 一貴, 菊地 利明, 松本 壮吉

    日本細菌学雑誌   76 ( 1 )   65 - 65   2021年2月

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    記述言語:日本語   出版者・発行元:日本細菌学会  

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  • 結核菌北京株のゲノム解析と組織透明化/3次元イメージング「CUBIC」による抗酸菌感染の生体内モニタリング

    袴田真理子, 袴田真理子, 瀧原速仁, 尾関百合子, 西山晃史, 立石善隆, 大橋璃子, 奥田修二郎, 田井中一貴, 菊地利明, 松本壮吉

    日本細菌学雑誌(Web)   76 ( 1 )   2021年

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  • 早期発症者と長期潜伏後発症者より分離した結核菌北京株のゲノム変異解析

    袴田真理子, 袴田真理子, 瀧原速仁, 岩本朋忠, 岩本朋忠, 田丸亜貴, 尾関百合子, 西山晃史, 立石善隆, 菊地利明, 奥田修二郎, 松本壮吉

    結核(Web)   96 ( 3 )   2021年

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  • 早期発症者と長期潜伏後発症者より分離した結核菌北京株のゲノム変異解析

    袴田 真理子, 瀧原 速仁, 岩本 朋忠, 田丸 亜貴, 尾関 百合子, 西山 晃史, 菊地 利明, 奥田 修二郎, 松本 壮吉

    結核   95 ( 5 )   115 - 115   2020年9月

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    記述言語:日本語   出版者・発行元:(一社)日本結核・非結核性抗酸菌症学会  

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  • MDP1はM.tuberculosis var BCG株の生存を確固とするための代謝と複製を調節する(MDP1 regulates metabolism and replication ensuring the survival of M. tuberculosis var BCG)

    シャバン・アミナ, 西山 晃史, 立石 善隆, 山口 雄大, 西内 由紀子, 瀧原 速仁, 奥田 修二郎, 松本 壮吉

    日本細菌学雑誌   75 ( 1 )   88 - 88   2020年1月

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    記述言語:英語   出版者・発行元:日本細菌学会  

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  • 早期発症者と長期潜伏後発症者より分離した結核菌北京株のゲノム変異についての解析

    袴田 真理子, 瀧原 速仁, 岩本 朋忠, 田丸 亜貴, 尾関 百合子, 西山 晃史, 立石 善隆, 菊地 利明, 奥田 修二郎, 松本 壮吉

    日本細菌学雑誌   75 ( 1 )   129 - 129   2020年1月

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    記述言語:日本語   出版者・発行元:日本細菌学会  

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  • 早期発症者と長期潜伏後発症者より分離した結核菌北京株のゲノム変異についての解析

    袴田真理子, 袴田真理子, 瀧原速仁, 岩本朋忠, 田丸亜貴, 尾関百合子, 西山晃史, 立石善隆, 菊地利明, 奥田修二郎, 松本壮吉

    日本細菌学雑誌(Web)   75 ( 1 )   2020年

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  • 早期発症者と長期潜伏後発症者より分離した結核菌北京株のゲノム変異解析

    袴田真理子, 袴田真理子, 瀧原速仁, 岩本朋忠, 田丸亜貴, 尾関百合子, 西山晃史, 菊地利明, 奥田修二郎, 松本壮吉

    結核(Web)   95 ( 5 )   2020年

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  • Rhodococcus属細菌の有機溶媒中での生育-アルカン相内での溶存酸素の利用について-

    岩淵範之, 田淵大樹, 瀧原速仁, 砂入道夫

    日本放線菌学会大会講演要旨集   34th   2019年

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  • カブトムシ幼虫の腸内環境と微生物群集構造について

    和田典子, 岩淵範之, 瀧原速仁, 奥田修二郎, 砂入道夫, 岩田隆太郎, 安齋寛

    日本微生物生態学会大会(Web)   33rd   2019年

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  • Rhodococcus属細菌の有機溶媒中での生育-アルカン相での溶存酸素の利用について-

    田淵大樹, 瀧原速仁, 岩淵範之, 砂入道夫

    日本ゲノム微生物学会年会要旨集   13th   2019年

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  • カブトムシ幼虫の腸内環境と微生物叢

    和田典子, 瀧原速仁, 奥田修二郎, 砂入道夫, 中嶋睦安, 岩淵範之, 安齋寛

    日本土壌肥料学会講演要旨集   64   2018年

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  • アルカン/培地二相培養系におけるRhodococcus erythropolis PR4の細胞局在性と酸素消費について

    田淵大樹, 瀧原速仁, 岩淵範之, 砂入道夫

    日本農芸化学会大会講演要旨集(Web)   2018   2018年

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  • Rhodococcus属細菌の有機溶媒耐性におけるgroEL2の影響

    岩淵範之, 瀧原速仁, 砂入道夫

    日本農芸化学会大会講演要旨集(Web)   2015   2015年

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  • アルカン相内で生育する有機溶媒耐性菌Phodococcus erythropolis PR4のタンパク質発現プロファイルの解析

    岩淵範之, 瀧原速仁, 奥田修二郎, 荻原淳, 砂入道夫

    日本土壌微生物学会講演要旨集   2014   2014年

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  • Rhodococcus erythropolis PR4株における細胞のアルカン相への転移に対するMgトランスポーターの影響

    瀧原速仁, 荻原淳, 岩淵範之, 砂入道夫

    日本農芸化学会大会講演要旨集(Web)   2013   2013年

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  • Rhodococcus erythropolis PR_4株のアルカン培地二相培養系における生育に対するMgSO_4の影響(PMバイオフィルム・界面,ポスター発表)

    瀧原速仁, 岩淵範之, 砂入道夫

    日本微生物生態学会講演要旨集   ( 28 )   2012年

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  • Rhodococcus rhodochrousの培地/アルカン二相培養系における細胞の局在性へのgroEL2遺伝子の影響

    松浦千明, 瀧原速仁, 岩淵範之, 砂入道夫

    日本生物工学会大会講演要旨集   64th   2012年

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  • 石油分解菌のユニークな性質(S12シンポジウムセッション)

    瀧原速仁, 岩淵範之, 砂入道夫

    日本微生物生態学会講演要旨集   ( 28 )   2012年

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  • Rhodococcus erythropolis PR4株の培地/アルカン二相培養系における細胞の局在性に対するGroEL2の影響

    瀧原速仁, 荻原淳, 吉田尊夫, 島村繁, 岩淵範之, 砂入道夫

    日本農芸化学会大会講演要旨集   2011   2011年

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  • Rhodococcus erythropolis PR4株の培地/アルカン二相培養系での細胞の局在性,溶媒耐性に対するGroEL2の影響

    瀧原速仁, 岩淵範之, 吉田尊夫, 島村繁, 砂入道夫, 荻原淳

    日本微生物生態学会講演要旨集   26th   2010年

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  • Rhodococcus erythropolis PR4株の培地/アルカン二相培養系における細胞の局在性に対するGroEL2の影響

    岩淵範之, 明瀬由美子, 瀧原速仁, 砂入道夫

    日本放線菌学会大会講演要旨集   25th   2010年

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  • Rhodococcus erythropolis PR4株の培地/アルカン二相培養系におけるプロテオーム解析

    瀧原速仁, 岩淵範之, 吉田尊夫, 島村繁, 砂入道夫, 荻原淳

    日本生物工学会大会講演要旨集   62nd   2010年

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  • Rhodococcus erythropolis PR4株の培地/アルカン二相培養系における各種プロテオーム解析

    瀧原速仁, 岩淵範之, 荻原淳, 吉田尊夫, 島村繁, 砂入道夫, 中嶋睦安, 中嶋睦安

    日本農芸化学会大会講演要旨集   2010   2010年

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講演・口頭発表等

  • Investigate carbohydrate metabolism in various environments using the distribution of glycan-related genes obtained from metagenomes 招待

    Hayato Takihara

    International Symposium on Microbial Glycoconjugates and the GlySpace Alliance: from micro to macro glycoscience  2022年3月 

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    開催年月日: 2022年3月

    記述言語:英語   会議種別:シンポジウム・ワークショップ パネル(指名)  

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  • IIBMP2022 WS4 医療から環境まで広がる糖鎖の世界へ 招待

    瀧原速仁

    千里ライフサイエンスセンター  2022年9月 

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    会議種別:シンポジウム・ワークショップ パネル(指名)  

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産業財産権

  • 蛍光物質及びその製造方法

    岩淵 範之, 松藤 寛, 砂入 道夫, 瀧原 速仁, 佐々木 太平, 白井 智也

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    出願人:学校法人日本大学

    出願番号:特願2014-167282  出願日:2014年8月

    公開番号:特開2016-044194  公開日:2016年4月

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  • 炭化水素の処理方法

    岩淵 範之, 中嶋 睦安, 砂入 道夫, 瀧原 速仁

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    出願人:学校法人日本大学

    出願番号:特願2009-206309  出願日:2009年9月

    公開番号:特開2011-055730  公開日:2011年3月

    特許番号/登録番号:特許第5532474号  登録日:2014年5月 

    J-GLOBAL

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  • 形質転換されたロドコッカス(Rhodococcus)属細菌、及び、これを用いた炭化水素の処理方法

    岩淵 範之, 中嶋 睦安, 砂入 道夫, 瀧原 速仁

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    出願人:学校法人日本大学

    出願番号:特願2009-206309  出願日:2009年9月

    公開番号:特開2011-055730  公開日:2011年3月

    J-GLOBAL

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