研究者詳細

顔写真

ニシヤマ　ケイタ
西山　啓太
Keita Nishiyama
所属
大学院農学研究科　生物生産科学専攻　動物生命科学講座（動物食品機能学分野）
職名
准教授
学位

  • 博士（農学）（北里大学）

  • 修士（農学）（北里大学）

経歴 4

  • 2022年10月 ～ 継続中
    東北大学大学院農学研究科　動物食品機能学分野　准教授

  • 2022年4月 ～ 2022年9月
    森永乳業株式会社　基礎研究所　研究員

  • 2019年4月 ～ 2022年3月
    慶應義塾大学　医学部 微生物学・免疫学教室　講師

  • 2015年4月 ～ 2019年3月
    北里大学　薬学部 微生物学教室　助教

学歴 1

  • 北里大学大学院　獣医学系研究科 博士課程修了

    2015年3月 ～

委員歴 2

  • Bioscience of Microbiota, Food and Health　編集副委員長

    2023年 ～ 継続中

  • 日本乳酸菌学会　理事、副編集長

    2020年 ～ 継続中

所属学協会 5

  • 日本農芸化学会

    2012年 ～ 継続中

  • 日本乳酸菌学会

    2010年 ～ 継続中

  • 日本細菌学会

    2015年 ～

  • 日本酪農科学会

    2010年 ～

  • 日本畜産学会

    2010年 ～

研究キーワード 5

  • シアル酸

  • ムチン

  • 消化管定着

  • 乳酸菌

  • 腸内細菌

研究分野 2

  • ライフサイエンス / 動物生産科学 /

  • ライフサイエンス / 応用微生物学 /

受賞 2

  1. 第 36 回 二宮善基記念賞

    2021年6月　学校法人北里研究所

  2. 日本酪農科学会 奨励賞

    2017年9月　日本酪農科学会　乳酸菌およびビフィズス菌の消化管定着性に関する研究

論文 48

  1. Milk sialyl-oligosaccharides mediate the early colonization of gut commensal microbes in piglets. 国際誌 査読有り

    Ryoga Hashimoto, Keita Nishiyama, Fu Namai, Kasumi Suzuki, Taiga Sakuma, Itsuko Fukuda, Yuta Sugiyama, Kenji Okano, Takafumi Shanoh, Eita Toyoshi, Ryusuke Ohgi, Sudeb Saha, Sae Tsuchida, Eri Nishiyama, Takao Mukai, Mutsumi Furukawa, Tomonori Nochi, Julio Villena, Wakako Ikeda-Ohtsubo, Gou Yoshioka, Eri Nakazaki, Yoshihito Suda, Haruki Kitazawa

    Microbiome　13　(1)　135-135　2025年5月24日

    DOI： 10.1186/s40168-025-02129-3 　

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    BACKGROUND: The suckling period in pigs is a key phase in development for shaping the gut microbiota, which is essential for maintaining biological homeostasis in neonates. In piglets fed sow milk, the gut microbiota comprises predominantly lactobacilli, indicating a host-gut microbiota symbiosis that is influenced by sow milk components. In this study, we sought to elucidate the mechanisms underlying the establishment and maintenance of the gut microbiome in suckling piglets, with a specific focus on the metabolism of sialyl-oligosaccharides by lactobacilli. RESULTS: Based on liquid chromatography-mass spectrometry analysis, we identified 3'-sialyl-lactose (3'SL) as the major oligosaccharide in porcine milk, and microbiome profiling revealed the predominance of Ligilactobacillus salivarius during the suckling period, with a subsequent transition to Limosilactobacillus reuteri dominance post-weaning. Notably, sialic acid metabolism was established to be exclusively attributable to L. salivarius, thereby highlighting the pivotal role of 3'SL in determining species-specific bacterial segregation. L. salivarius was found to metabolize 3'SL when co-cultured with Bacteroides thetaiotaomicron, resulting in a shift in the predominant short-chain fatty acid produced, from lactate to acetate. This metabolic shift, in turn, inhibits the growth of enterotoxigenic Escherichia coli. Furthermore, the comparison of the gut microbiota between suckling piglets and those fed a low-3'SL formula revealed distinct diversity profiles. We accordingly speculate that an absence of sialyl-oligosaccharides in the formula-fed piglets may have restricted the growth of sialic acid-utilizing bacteria such as L. salivarius, thereby leading to a higher abundance of Enterobacteriaceae. CONCLUSIONS: Our findings reveal the influence of sialyl-oligosaccharides in promoting microbial diversity and gut homeostasis, thereby highlighting the importance of sialic acid as a key factor in shaping milk-driven microbial colonization during the early stages of piglet development. Video Abstract.

  2. Modulation of Macrophages TLR4-Mediated Transcriptional Response by Lacticaseibacillus rhamnosus CRL1505 and Lactiplantibacillus plantarum CRL1506 国際誌 国際共著 査読有り

    Masahiko Suzuki†, Ayelen Baillo†, Leonardo Albarracin, Mariano Elean, Rodrigo Serda, Yoshihito Suda, Fu Namai, Keita Nishiyama, Haruki Kitazawa*, Julio Villena* (†: Equal contribution)

    Int. J. Mol. Sci.　26:2688　(6)　1-26　2025年3月17日

    出版者・発行元： MDPI

    DOI： 10.3390/ijms26062688 　

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    IF = 4.9

  3. Orally Administered Lactobacilli Strains Modulate Alveolar Macrophages and Improve Protection Against Respiratory Superinfection 国際誌 国際共著 査読有り

    Leonardo Albarracin, Stefania Dentice Maidana, Kohtaro Fukuyama, Mariano Elean, Julio Nicolás, Argañaraz Aybar, Yoshihito Suda, Keita Nishiyama, Haruki Kitazawa*, Julio Villena*

    Biomolecules　14:1600　(12)　1-17　2024年12月14日

    出版者・発行元： MDPI

    DOI： 10.3390/biom14121600 　

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    IF = 4.8

  4. Modulation of the Gut–Lung Axis by Water Kefir and Kefiran and Their Impact on Toll-like Receptor 3-Mediated Respiratory Immunity 国際誌 国際共著 査読有り

    Stefania Dentice Maidana†, Julio Nicolás Argañaraz Aybar†, Leonardo Albarracin, Yoshiya Imamura, Luciano Arellano-Arriagada, Fu Namai, Yoshihito Suda, Keita Nishiyama, Julio Villena*, Haruki Kitazawa* (†: Equal contribution)

    Biomolecules　14　(11)　2024年11月17日

    出版者・発行元： MDPI

    DOI： 10.3390/biom14111457 　

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    IF = 4.8

  5. Role of immunomodulatory probiotics in alleviating bacterial diarrhea in piglets: a systematic review 国際誌 国際共著 査読有り

    Sudeb Saha, Fu Namai, Keita Nishiyama, Julio Villena*, Haruki Kitazawa*

    J Anim Sci Biotechnol　15:112　(1)　1-25　2024年8月12日

    出版者・発行元： Springer Nature

    DOI： 10.1186/s40104-024-01070-z 　

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    IF = 6.3

  6. Exploring strain-level diversity in the gut microbiome through mucin particle adhesion. 国際誌

    Keita Nishiyama, Ryuta Murakami, Masaki Nakahata, Binghui Zhou, Nanami Hashikura, Hiroki Kaneko, Fu Namai, Wakako Ikeda-Ohtsubo, Jin-Zhong Xiao, Haruki Kitazawa, Toshitaka Odamaki

    Applied and environmental microbiology　90　(9)　e0123524　2024年8月12日

    DOI： 10.1128/aem.01235-24 　

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    UNLABELLED: Mucin glycoproteins are a significant source of carbon for the gut bacteria. Various gut microbial species possess diverse hydrolytic enzymes and catabolic pathways for breaking down mucin glycans, resulting in competition for the limited nutrients within the gut environment. Adherence to mucin glycans represents a crucial strategy used by gut microbes to access nutrient reservoirs. Understanding these properties is pivotal for comprehending the survival mechanisms of bacteria in the gastrointestinal tract. However, characterization of individual strains within the vast array of coexisting bacteria in the microbiome is challenging. To investigate this, we developed mucin-immobilized particles by immobilizing porcine gastric mucin (PGM) onto glass beads chemically modified with boronic acid. These PGM-immobilized particles were then anaerobically cultured with human fecal microbiota, and the bacteria adhering to PGM were isolated. Interestingly, the microbiome composition remained largely unchanged irrespective of PGM immobilization. Nonetheless, bacteria isolated from PGM-immobilized glass particles exhibited notably higher N-acetylgalactosaminidase activity compared to the control beads. Furthermore, Bacteroides strains isolated from PGM-immobilized glass particles displayed enhanced adhesive and metabolic properties to PGM. These findings underscore the utility of PGM particles in enriching and isolating specific microbes. Moreover, they highlight substantial differences in microbial properties at the strain level. We anticipate that PGM-immobilized particles will advance culture-based microbiome research, emphasizing the significance of strain-level characterization. IMPORTANCE: Metabolism of mucin glycans by gut bacteria represents a crucial strategy for accessing nutrient reservoirs. The efficacy of mucin glycan utilization among gut bacteria hinges on the metabolic capabilities of individual strains, necessitating meticulous strain-level characterization. In this investigation, we used glass beads chemically immobilized with mucins to selectively enrich bacteria from fecal fermentation cultures, based on their superior adhesion to and metabolism of mucin glycoproteins. These findings lend support to the hypothesis that the physical interactions between bacteria and mucin glycoprotein components directly correlate with their capacity to utilize mucins as nutrient sources. Furthermore, our study implies that physical proximity may significantly influence bacterial nutrient acquisition within the ecosystem, facilitating gut bacteria's access to carbohydrate components.

  7. Modulation of the Toll-like Receptor 3-Mediated Intestinal Immune Response by Water Kefir 国際誌 国際共著 査読有り

    Stefania Dentice Maidana†, Ramiro Ortiz Moyano†, Mariano Elean, Yoshiya Imamura, Leonardo Albarracín, Fu Namai, Yoshihito Suda, Keita Nishiyama, Julio Villena*, Haruki Kitazawa*, (†: Equal contribution)contribution

    Microbiol. Res.　15　1239-1250　2024年7月20日

    出版者・発行元： MDPI

    DOI： 10.3390/microbiolres15030083 　

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    IF=2.1

  8. Antagonistic effects of Corynebacterium pseudodiphtheriticum 090104 on Respiratory Pathogens 国際誌 国際共著 査読有り

    Ramiro Ortiz Moyano†, Stefania Dentice Maidana†, Yoshiya Imamura, Mariano Elean, Fu Namai, Yoshihito Suda, Keita Nishiyama, Vyacheslav Melnikov, Haruki Kitazawa*, Julio Villena* (†: Equal contribution)

    Microorganisms　12:1295　(7)　1-17　2024年6月26日

    出版者・発行元： MDPI

    DOI： 10.3390/microorganisms12071295 　

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    IF = 4.1

  9. The Mucus-Binding Factor Mediates Lacticaseibacillus rhamnosus CRL1505 Adhesion but not Immunomodulation in the Respiratory Tract 国際誌 国際共著

    Binghui Zhou†, Mariano Elean†, Lorena Arce, Kohtaro Fukuyama, Kae Tomotsune, Stefania Dentice Maidana, Sudeb Saha, Fu Namai, Keita Nishiyama, María Guadalupe Vizoso-Pinto, Julio Villena*, Haruki Kitazawa* (†: Equal contribution)

    Microorganisms　12:1209　(6)　1-12　2024年6月16日

    出版者・発行元： MDPI

    DOI： 10.3390/microorganisms12061209 　

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    IF = 4.5

  10. Insights into the Anti-Adipogenic and Anti-Inflammatory Potentialities of Probiotics against Obesity 国際誌 国際共著 査読有り

    A. K. M. Humayun Kober, Sudeb Saha, Mutamed M Ayyash, Fu Namai, Keita Nishiyama, Kazutoyo Yoda, Julio Villena, Haruki Kitazawa

    Nutrients　16:1373　(9)　1-24　2024年4月30日

    出版者・発行元： MDPI

    DOI： 10.3390/nu16091373 　

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    IF = 5.9

  11. Effect of Lactiplantibacillus plantarum cell-free culture on bacterial pathogens isolated from cystic fibrosis patients: in vitro and in vivo studies. 国際誌

    Carla Luciana Abán, Silvia Orosco, Julio Nicolás Argañaraz Aybar, Leonardo Albarracín, Analía Venecia, Liliana Perret, Sonia Ortiz Mayor, Keita Nishiyama, Juan Carlos Valdéz, Haruki Kitazawa, Julio Villena, Nadia Gobbato

    Frontiers in microbiology　15　1440090-1440090　2024年

    DOI： 10.3389/fmicb.2024.1440090 　

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    This study aimed to investigate the effects of the cell-free supernatant of Lactiplantibacillus plantarum ATCC® 10241TM on the biofilm-forming capacity of Pseudomonas aeruginosa strains isolated from cystic fibrosis (CF) patients. In addition, the study evaluated the in vivo potential of the cell-free supernatant to modulate inflammation and reduce lung damage in mice infected with P. aeruginosa strains or co-challenged with P. aeruginosa and the Streptococcus milleri group (SMG). The results showed that CF-derived P. aeruginosa strains can infect the respiratory tract of adult mice, inducing local inflammation and lung damage. The severity of these infections was exacerbated when P. aeruginosa was co-administered with SMG. Notably, nebulization with the cell-free supernatant of L. plantarum produced beneficial effects, reducing respiratory infection severity and inflammatory responses induced by P. aeruginosa, both alone or in combination with SMG. Reduced bacterial loads and lung damage were observed in supernatant-treated mice compared to controls. Although further mechanistic studies are necessary, the results show that the cell-free supernatant of L. plantarum ATCC® 10241TM is an interesting adjuvant alternative to treat P. aeruginosa respiratory infections and superinfections in CF patients.

  12. Adhesion Inhibition Assay for Helicobacter pylori to Mucin by Lactobacillus. 国際誌

    Keita Nishiyama, Takao Mukai

    Methods in molecular biology (Clifton, N.J.)　2763　353-358　2024年

    DOI： 10.1007/978-1-0716-3670-1_30 　

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    The ability of Lactobacillus to adhere to mucin is a parameter for evaluating the effectiveness of probiotics. In particular, a competitive inhibition assay of pathogenic bacteria using mucin-adherent lactobacilli is useful for identifying Lactobacillus strains capable of preventing mucus from being colonized by pathogens. Here, we describe an adhesion inhibition assay method for Helicobacter pylori to porcine gastric mucin by Limosilactobacillus reuteri.

  13. Development of an intestinal epithelial cell line and organoids derived from the same swine and characterization of their antiviral responses 査読有り

    Kaho MATSUMOTO, Fu NAMAI, Ayako MIYAZAKI, Yoshiya IMAMURA, Kohtaro FUKUYAMA, Wakako IKEDA-OHTSUBO, Keita NISHIYAMA, Julio VILLENA, Kohtaro MIYAZAWA, Haruki KITAZAWA

    Bioscience of Microbiota, Food and Health　43　(4)　342-351　2024年

    出版者・発行元： BMFH Press

    DOI： 10.12938/bmfh.2024-0046 　

    eISSN：2186-3342

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    Intestinal homeostasis and integrity are important factors for maintaining host health. This study established intestinal epithelial cell lines and organoids from the same swine jejunal crypts to develop seamless swine intestinal in vitro evaluation systems. The study evaluated the proliferative capacity and tight junction formation of the epithelial cell line and characterized the cell differentiation potential of the intestinal organoids. The evaluation systems were subsequently exposed to the Toll-like receptor 3 (TLR3) agonist poly(I:C) to simulate viral infections and assess the antiviral responses. The results demonstrated no differences in the response to type I interferons. There were, however, significant differences in the expression of interferon-stimulated genes. This study collectively introduced a flexible evaluation system using cell lines and organoids and revealed notable differences in the expression of interferon-stimulated genes, highlighting the complexity of the immune responses in these in vitro systems and the importance of intestinal heterogeneity in assessing viral responses.

  14. In vitro evaluation of the immunomodulatory and wakame assimilation properties of Lactiplantibacillus plantarum strains from swine milk. 国際誌 査読有り

    Ryusuke Ohgi, Sudeb Saha, Binghui Zhou, Taiga Sakuma, Mitsuki Sakurai, Yuhka Nakano, Fu Namai, Wakako Ikeda-Ohtsubo, Yoshihito Suda, Keita Nishiyama, Julio Villena, Haruki Kitazawa

    Frontiers in microbiology　15　1324999-1324999　2024年

    DOI： 10.3389/fmicb.2024.1324999 　

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    The emergence and spread of antibiotic resistance threat forced to explore alternative strategies for improving the resistance to pathogens in livestock production. Probiotic lactic acid bacteria represent an alternative for this objective. In this study, seven Lactiplantibacillus plantarum strains from porcine colostrum and milk were isolated, identified and characterized in terms of their abilities to modulate immunity in porcine intestinal epithelial (PIE) cells. Then, two potential immunoregulatory strains were studied in terms of their ability to utilize and grow in wakame (Undaria pinnafida). Isolates were identified by 16S rRNA gene and evaluated by studying their interaction with PIE cells. The expressions of peptidoglycan recognition proteins (PGRPs), nucleotide-binding oligomerization domain (NODs), host defense peptides (pBD), and type I interferons (IFNs) were evaluated by RT-qPCR. The strain 4M4417 showed a remarkable capacity to differentially regulate the expression of PGRP1, PGRP3, NOD1, NOD2, and pBD1 in PIE cells. On the other hand, the strain 4M4326 was the most efficient to improve the expression of IFN-α and IFN-β in PIE cells challenged with poly (I:C). Both L. plantarum 4M4326 and 4M4417 were characterized in terms of their ability to utilize wakame. Results demonstrated that both strains efficiently grew in wakame-based broth. Our results suggest that L. planatrum 4M4326 and 4M4417 are interesting candidates to develop immunomodulatory feeds based on wakame utilization. These new immunosynbiotic feeds could help to reduce severity of intestinal infections and improve immune health status in pigs.

  15. Immunobiotic Ligilactobacillus salivarius FFIG58 Confers Long-Term Protection against Streptococcus pneumoniae 国際誌 国際共著 査読有り

    Mariano Elean, Fernanda Raya-Tonetti, Kohtaro Fukuyama, Luciano Arellano-Arriagada, Fu Namai, Yoshihito Suda, Nadia Gobbato, Keita Nishiyama, Julio Villena, Haruki Kitazawa

    International Journal of Molecular Sciences　24(21):15773　(21)　1-16　2023年10月30日

    出版者・発行元： MDPI

    DOI： 10.3390/ijms242115773 　

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    IF = 5.6

  16. D-amino Acids Ameliorate Experimental Colitis and Cholangitis by Inhibiting Growth of Proteobacteria: Potential Therapeutic Role in Inflammatory Bowel Disease. 国際誌 査読有り

    Satoko Umeda, Tomohisa Sujino, Kentaro Miyamoto, Yusuke Yoshimatsu, Yosuke Harada, Keita Nishiyama, Yoshimasa Aoto, Keika Adachi, Naoki Hayashi, Kimiko Amafuji, Nobuko Moritoki, Shinsuke Shibata, Nobuo Sasaki, Masashi Mita, Shun Tanemoto, Keiko Ono, Yohei Mikami, Jumpei Sasabe, Kaoru Takabayashi, Naoki Hosoe, Toshihiko Suzuki, Toshiro Sato, Koji Atarashi, Toshiaki Teratani, Haruhiko Ogata, Nobuhiro Nakamoto, Daisuke Shiomi, Hiroshi Ashida, Takanori Kanai

    Cellular and molecular gastroenterology and hepatology　16　(6)　1011-1031　2023年8月9日

    DOI： 10.1016/j.jcmgh.2023.08.002 　

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    BACKGROUND & AIMS: D-amino acids, the chiral counterparts of protein L-amino acids, were primarily produced and utilized by microbes, including those in the human gut. However, little was known about how orally administered or microbe-derived D-amino acids affected the gut microbial community or gut disease progression. METHODS: The ratio of D- to L-amino acids was analyzed in feces and blood from patients with ulcerative colitis (UC) and healthy controls. Also, composition of microbe was analyzed from patients with UC. Mice were treated with D-amino acid in dextran sulfate sodium colitis model and liver cholangitis model. RESULTS: The ratio of D- to L-amino acids was lower in the feces of patients with UC than that of healthy controls. Supplementation of D-amino acids ameliorated UC-related experimental colitis and liver cholangitis by inhibiting growth of Proteobacteria. Addition of D-alanine, a major building block for bacterial cell wall formation, to culture medium inhibited expression of the ftsZ gene required for cell fission in the Proteobacteria Escherichia coli and Klebsiella pneumoniae, thereby inhibiting growth. Overexpression of ftsZ restored growth of E. coli even when D-alanine was present. We found that D-alanine not only inhibited invasion of pathological K. pneumoniae into the host via pore formation in intestinal epithelial cells but also inhibited growth of E. coli and generation of antibiotic-resistant strains. CONCLUSIONS: D-amino acids might have potential for use in novel therapeutic approaches targeting Proteobacteria-associated dysbiosis and antibiotic-resistant bacterial diseases by means of their effects on the intestinal microbiota community.

  17. Recent Advances in the Use of Probiotics to Improve Meat Quality of Small Ruminants: A Review 国際誌

    Sudeb Saha, Kohtaro Fukuyama, Marina Debnath, Fu Namai, Keita Nishiyama, Haruki Kitazawa

    Microorganisms　11　(7)　1652-1652　2023年6月25日

    出版者・発行元： MDPI AG

    DOI： 10.3390/microorganisms11071652 　

    eISSN：2076-2607

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    Meat from small ruminants is considered a high quality and delicacy product in many countries. Several benefits have been perceived from probiotics as dietary supplements, such as improved carcass weight, color, tenderness, flavor, muscle fiber structure, water-holding capacity, and healthy fatty acid profile of the meat. Thus, the present review focuses on the effect of probiotics on improving the quality of meat from small ruminants. Though many benefits have been associated with the use of probiotics, the findings of all the considered articles are not always consistent, and the mechanisms behind improving meat quality are not appropriately defined. This variability of findings could be due to the use of different probiotic strains, dosage rates, number of days of experiment, nutrition, breed, age, and health status of the animals. Therefore, future research should emphasize specific strains, optimal dose and days of administration, route, and mechanisms for the specific probiotic strains to host. This review provides a comprehensive overview of the use of probiotics for small ruminants and their impact on meat quality.

  18. Partial Characterization and Immunomodulatory Effects of Exopolysaccharides from Streptococcus thermophilus SBC8781 during Soy Milk and Cow Milk Fermentation. 国際誌 査読有り

    Hajime Nakata, Yoshiya Imamura, Sudeb Saha, René Emanuel Lobo, Shugo Kitahara, Shota Araki, Mikado Tomokiyo, Fu Namai, Masanori Hiramitsu, Takashi Inoue, Keita Nishiyama, Julio Villena, Haruki Kitazawa

    Foods (Basel, Switzerland)　12　(12)　2023年6月15日

    DOI： 10.3390/foods12122374 　

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    The immunomodulatory properties of exopolysaccharides (EPSs) produced by Streptococcus thermophilus have not been explored in depth. In addition, there are no comparative studies of the functional properties of EPSs produced by streptococci in different food matrices. In this work, EPSs from S. thermophilus SBC8781 were isolated after soy milk (EPS-s) or cow milk (EPS-m) fermentation, identified, and characterized in their abilities to modulate immunity in porcine intestinal epithelial cells. Fresh soy milk and cow milk were inoculated with S. thermophilus SBC8781 (7 log CFU/mL) and incubated at 37 °C for 24 h. The extraction of EPSs was performed by the ethanol precipitation method. Analytical techniques, including NMR, UV-vis spectroscopy, and chromatography, identified and characterized both biopolymer samples as polysaccharides with high purity levels and similar Mw. EPS-s and EPS-m had heteropolysaccharide structures formed by galactose, glucose, rhamnose, ribose, and mannose, although with different monomer proportions. On the other hand, EPS-s had higher quantities of acidic polymer than EPS-m. The biopolymer production of the SBC8781 strain from the vegetable culture broth was 200-240 mg/L, which was higher than that produced in milk, which reached concentrations of 50-70 mg/L. For immunomodulatory assays, intestinal epithelial cells were stimulated with 100 µg/mL of EPS-s or EPS-m for 48 h and then stimulated with the Toll-like receptor 3 agonist poly(I:C). EPS-s significantly reduced the expression of IL-6, IFN-β, IL-8, and MCP-1 and increased the negative regulator A20 in intestinal epithelial cells. Similarly, EPS-m induced a significant reduction of IL-6 and IL-8 expressions, but its effect was less remarkable than that caused by EPS-s. Results indicate that the structure and the immunomodulatory activity of EPSs produced by the SBC8781 strain vary according to the fermentation substrate. Soy milk fermented with S. thermophilus SBC8781 could be a new immunomodulatory functional food, which should be further evaluated in preclinical trials.

  19. Oral Administration of Lacticaseibacillus rhamnosus CRL1505 Modulates Lung Innate Immune Response against Klebsiella pneumoniae ST25 国際誌 国際共著 査読有り

    Stefania Dentice Maidana†, Yoshiya Imamura†, Mariano Elean, Leonardo Albarracín, Keita Nishiyama, Yoshihito Suda, Shoichiro Kurata, María Ángela Jure, Haruki Kitazawa, Julio Villena, Equal contribution

    Microorganisms　11:1148　(5)　1-13　2023年4月28日

    出版者・発行元： MDPI

    DOI： 10.3390/microorganisms11051148 　

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    IF = 4.926

  20. A bacterial sulfoglycosidase highlights mucin O-glycan breakdown in the gut ecosystem. 国際誌 査読有り

    Toshihiko Katoh, Chihaya Yamada, Michael D Wallace, Ayako Yoshida, Aina Gotoh, Moe Arai, Takako Maeshibu, Toma Kashima, Arno Hagenbeek, Miriam N Ojima, Hiromi Takada, Mikiyasu Sakanaka, Hidenori Shimizu, Keita Nishiyama, Hisashi Ashida, Junko Hirose, Maria Suarez-Diez, Makoto Nishiyama, Ikuo Kimura, Keith A Stubbs, Shinya Fushinobu, Takane Katayama

    Nature chemical biology　19　(6)　778-789　2023年3月2日

    DOI： 10.1038/s41589-023-01272-y 　

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    Mucinolytic bacteria modulate host-microbiota symbiosis and dysbiosis through their ability to degrade mucin O-glycans. However, how and to what extent bacterial enzymes are involved in the breakdown process remains poorly understood. Here we focus on a glycoside hydrolase family 20 sulfoglycosidase (BbhII) from Bifidobacterium bifidum, which releases N-acetylglucosamine-6-sulfate from sulfated mucins. Glycomic analysis showed that, in addition to sulfatases, sulfoglycosidases are involved in mucin O-glycan breakdown in vivo and that the released N-acetylglucosamine-6-sulfate potentially affects gut microbial metabolism, both of which were also supported by a metagenomic data mining analysis. Enzymatic and structural analysis of BbhII reveals the architecture underlying its specificity and the presence of a GlcNAc-6S-specific carbohydrate-binding module (CBM) 32 with a distinct sugar recognition mode that B. bifidum takes advantage of to degrade mucin O-glycans. Comparative analysis of the genomes of prominent mucinolytic bacteria also highlights a CBM-dependent O-glycan breakdown strategy used by B. bifidum.

  21. Sharing of Moonlighting Proteins Mediates the Symbiotic Relationship among Intestinal Commensals. 国際誌 査読有り

    Keita Nishiyama, Cheng-Chung Yong, Nobuko Moritoki, Haruki Kitazawa, Toshitaka Odamaki, Jin-Zhong Xiao, Takao Mukai

    Applied and environmental microbiology　89　(3)　e0219022　2023年2月27日

    DOI： 10.1128/aem.02190-22 　

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    The human gastrointestinal tract is inhabited by trillions of symbiotic bacteria that form a complex ecological community and influence human physiology. Symbiotic nutrient sharing and nutrient competition are the most studied relationships in gut commensals, whereas the interactions underlying homeostasis and community maintenance are not fully understood. Here, we provide insights into a new symbiotic relationship wherein the sharing of secreted cytoplasmic proteins, called "moonlighting proteins," between two heterologous bacterial strains (Bifidobacterium longum and Bacteroides thetaiotaomicron) was observed to affect the adhesion of bacteria to mucins. B. longum and B. thetaiotaomicron were cocultured using a membrane-filter system, and in this system the cocultured B. thetaiotaomicron cells showed greater adhesion to mucins compared to that shown by monoculture cells. Proteomic analysis showed the presence of 13 B. longum-derived cytoplasmic proteins on the surface of B. thetaiotaomicron. Moreover, incubation of B. thetaiotaomicron with the recombinant proteins GroEL and elongation factor Tu (EF-Tu)-two well-known mucin-adhesive moonlighting proteins of B. longum-led to an increase in the adhesion of B. thetaiotaomicron to mucins, a result attributed to the localization of these proteins on the B. thetaiotaomicron cell surface. Furthermore, the recombinant EF-Tu and GroEL proteins were observed to bind to the cell surface of several other bacterial species; however, the binding was species dependent. The present findings indicate a symbiotic relationship mediated by the sharing of moonlighting proteins among specific strains of B. longum and B. thetaiotaomicron. IMPORTANCE The adhesion of intestinal bacteria to the mucus layer is an important colonization strategy in the gut environment. Generally, the bacterial adhesion process is a characteristic feature of the individual cell surface-associated adhesion factors secreted by a particular bacterium. In this study, coculture experiments between Bifidobacterium and Bacteroides show that the secreted moonlighting proteins adhere to the cell surface of coexisting bacteria and alter the adhesiveness of the bacteria to mucins. This finding indicates that the moonlighting proteins act as adhesion factors for not only homologous strains but also for coexisting heterologous strains. The presence of a coexisting bacterium in the environment can significantly alter the mucin-adhesive properties of another bacterium. The findings from this study contribute to a better understanding of the colonization properties of gut bacteria through the discovery of a new symbiotic relationship between them.

  22. Establishment of porcine fecal-derived ex vivo microbial communities to evaluate the impact of livestock feed on gut microbiome 査読有り

    Yuji TSUJIKAWA, Keita NISHIYAMA, Fu NAMAI, Yoshiya IMAMURA, Taiga SAKUMA, Sudeb SAHA, Masahiko SUZUKI, Mitsuki SAKURAI, Ryo IWATA, Kengo MATSUO, Hironori TAKAMORI, Yoshihito SUDA, Binghui ZHOU, Itsuko FUKUDA, Julio VILLENA, Iwao SAKANE, Ro OSAWA, Haruki KITAZAWA

    Bioscience of Microbiota, Food and Health　43　(2)　100-109　2023年

    出版者・発行元： BMFH Press

    DOI： 10.12938/bmfh.2023-085 　

    eISSN：2186-3342

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    Sustainable livestock production requires reducing competition for food and feed resources and increasing the utilization of food by-products in livestock feed. This study describes the establishment of an anaerobic batch culture model to simulate pig microbiota and evaluate the effects of a food by-product, wakame seaweed stalks, on ex vivo microbial communities. We selected one of the nine media to support the growth of a bacterial community most similar in composition and diversity to that observed in pig donor feces. Supplementation with wakame altered the microbial profile and short-chain fatty acid composition in the ex vivo model, and a similar trajectory was observed in the in vivo pig experimental validation. Notably, the presence of wakame increased the abundance of Lactobacillus species, which may have been due to cross-feeding with Bacteroides. These results suggest the potential of wakame as a livestock feed capable of modulating the pig microbiome. Collectively, this study highlights the ability to estimate the microbiome changes that occur when pigs are fed a specific feed using an ex vivo culture model.

  23. Establishment of a porcine bronchial epithelial cell line and its application to study innate immunity in the respiratory epithelium. 国際誌 査読有り

    Kohtaro Fukuyama, Tao Zhuang, Eita Toyoshi, Fernanda Raya Tonetti, Sudeb Saha, Binghui Zhou, Wakako Ikeda-Ohtsubo, Keita Nishiyama, Hisashi Aso, Julio Villena, Haruki Kitazawa

    Frontiers in immunology　14　1117102-1117102　2023年

    DOI： 10.3389/fimmu.2023.1117102 　

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    In vitro culture models that precisely mirror the porcine respiratory epithelium are needed to gain insight into how pathogens and host interact. In this study, a new porcine bronchial epithelial cell line, designated as PBE cells, was established from the respiratory tract of a neonatal pig. PBE cells assumed a cobblestone-epithelial like morphology with close contacts between the cells when they reached confluence. The PBE cell line was characterized in terms of its expression of pattern recognition receptors (PRRs) and its ability to respond to the activation of the Toll-like receptor 3 (TLR3) and TLR4 signaling pathways, which are key PRRs involved in the defense of the respiratory epithelium against pathogens. PBE cells stimulated with poly(I:C) were able to up-regulate the expression of IFN-β, IFN-λ1 (IL-29), IFN-λ3 (IL-28B), the antiviral factors Mx1, OAS1, and PKR, as well as the viral PRRs RIG-1 and MDA5. The expression kinetics studies of immune factors in PBE cells allow us to speculate that this cell line can be a useful in vitro tool to investigate treatments that help to potentiate antiviral immunity in the respiratory epithelium of the porcine host. In addition, poly(I:C) and LPS treatments increased the expression of the inflammatory cytokines TNF-α, IL-6, IL-8, and MCP-1/CCL2 and differentially modulated the expression of negative regulators of the TLR signaling pathways. Then, PBE cells may also allow the evaluation of treatments that can regulate TLR3- and TLR4-mediated inflammatory injury in the porcine airway, thereby protecting the host against harmful overresponses.

  24. The Mucus Binding Factor Is Not Necessary for Lacticaseibacillus rhamnosus CRL1505 to Exert Its Immunomodulatory Activities in Local and Distal Mucosal Sites. 国際誌 査読有り

    Kae Tomotsune, Fernanda Raya Tonetti, Hiroya Mizuno, Mariano Elean, Kohtaro Fukuyama, Binghui Zhou, Wakako Ikeda-Ohtsubo, Keita Nishiyama, Akihiro Yamamura, Hideaki Karasawa, Shinobu Ohnuma, Akira Horii, Tadao Saito, Haruki Kitazawa, Julio Villena

    International journal of molecular sciences　23　(22)　2022年11月18日

    DOI： 10.3390/ijms232214357 　

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    Both viable and non-viable orally administered Lacticaseibacillus rhamnosus CRL1505 modulate immunity in local (intestine) and distal (respiratory) mucosal sites. So, intestinal adhesion and colonization are not necessary for this probiotic strain to exert its immunomodulatory effects. In this work, a mucus-binding factor knockout CRL1505 strain (ΔmbfCRL1505) was obtained and the lack of binding ability to both intestinal epithelial cells and mucin was demonstrated in vitro. In addition, two sets of in vivo experiments in 6-week-old Balb/c mice were performed to evaluate ΔmbfCRL1505 immunomodulatory activities. (A) Orally administered ΔmbfCRL1505 prior to intraperitoneal injection of the Toll-like receptor 3 (TLR3) agonist poly(I:C) significantly reduced intraepithelial lymphocytes (CD3+NK1.1+CD8αα+) and pro-inflammatory mediators (TNF-α, IL-6 and IL-15) in the intestinal mucosa. (B) Orally administered ΔmbfCRL1505 prior to nasal stimulation with poly(I:C) significantly decreased the levels of the biochemical markers of lung tissue damage. In addition, reduced recruitment of neutrophils and levels of pro-inflammatory mediators (TNF-α, IL-6 and IL-8) as well as increased IFN-β and IFN-γ in the respiratory mucosa were observed in ΔmbfCRL1505-treated mice when compared to untreated control mice. The immunological changes induced by the ΔmbfCRL1505 strain were not different from those observed for the wild-type CRL1505 strain. Although it is generally accepted that the expression of adhesion factors is necessary for immunobiotics to induce their beneficial effects, it was demonstrated here that the mbf protein is not required for L. rhamnosus CRL1505 to exert its immunomodulatory activities in local and distal mucosal sites. These results are a step forward towards understanding the mechanisms involved in the immunomodulatory capabilities of L. rhamnosus CRL1505.

  25. Identification of trypsin-degrading commensals in the large intestine. 国際誌 査読有り

    Youxian Li, Eiichiro Watanabe, Yusuke Kawashima, Damian R Plichta, Zhujun Wang, Makoto Ujike, Qi Yan Ang, Runrun Wu, Munehiro Furuichi, Kozue Takeshita, Koji Yoshida, Keita Nishiyama, Sean M Kearney, Wataru Suda, Masahira Hattori, Satoshi Sasajima, Takahiro Matsunaga, Xiaoxi Zhang, Kazuto Watanabe, Jun Fujishiro, Jason M Norman, Bernat Olle, Shutoku Matsuyama, Ho Namkoong, Yoshifumi Uwamino, Makoto Ishii, Koichi Fukunaga, Naoki Hasegawa, Osamu Ohara, Ramnik J Xavier, Koji Atarashi, Kenya Honda

    Nature　609　(7927)　582-589　2022年9月

    DOI： 10.1038/s41586-022-05181-3 　

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    Increased levels of proteases, such as trypsin, in the distal intestine have been implicated in intestinal pathological conditions1-3. However, the players and mechanisms that underlie protease regulation in the intestinal lumen have remained unclear. Here we show that Paraprevotella strains isolated from the faecal microbiome of healthy human donors are potent trypsin-degrading commensals. Mechanistically, Paraprevotella recruit trypsin to the bacterial surface through type IX secretion system-dependent polysaccharide-anchoring proteins to promote trypsin autolysis. Paraprevotella colonization protects IgA from trypsin degradation and enhances the effectiveness of oral vaccines against Citrobacter rodentium. Moreover, Paraprevotella colonization inhibits lethal infection with murine hepatitis virus-2, a mouse coronavirus that is dependent on trypsin and trypsin-like proteases for entry into host cells4,5. Consistently, carriage of putative genes involved in trypsin degradation in the gut microbiome was associated with reduced severity of diarrhoea in patients with SARS-CoV-2 infection. Thus, trypsin-degrading commensal colonization may contribute to the maintenance of intestinal homeostasis and protection from pathogen infection.

  26. O-acetylesterase activity of Bifidobacterium bifidum sialidase facilities the liberation of sialic acid and encourages the proliferation of sialic acid scavenging Bifidobacterium breve. 国際誌 査読有り

    Tatsunari Yokoi, Keita Nishiyama, Yuka Kushida, Kazuya Uribayashi, Takahiro Kunihara, Rika Fujimoto, Yuji Yamamoto, Masahiro Ito, Tsuyoshi Miki, Takeshi Haneda, Takao Mukai, Nobuhiko Okada

    Environmental microbiology reports　14　(4)　637-645　2022年8月

    DOI： 10.1111/1758-2229.13083 　

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    Bifidobacterium bifidum possesses two extracellular sialidases (SiaBb1 and SiaBb2) that release free sialic acid from mucin sialoglycans, which can be utilized via cross-feeding by Bifidobacterium breve that, otherwise, is prevented from utilizing this nutrient source. Modification of sialic acids with O-acetyl esters is known to protect mucin glycans from degradation by bacterial sialidases. Compared to SiaBb2, SiaBb1 has an additional O-acetylesterase (Est) domain. We aimed to elucidate the role of the SiaBb1 Est domain from B. bifidum in sialic acid cross-feeding within Bifidobacterium. Pre-treatment of mucin secreted from bovine submaxillary glands (BSM) using His6 -tagged-Est and -SiaBb2 released a higher amount of sialic acid compared to the pre-treatment by His6 -SiaBb2. Growth of B. breve increased with an increase in nanE expression when supplemented with both His6 -Est- and His6 -SiaBb2-treated BSM. These results indicate that the esterase activity of the SiaBb1 Est domain enhances the efficiency of SiaBb2 to cleave sialic acid from mucin. This free sialic acid can be utilized by coexisting sialic acid scavenging B. breve via cross-feeding. Here, we provide the molecular mechanism underlying the unique sialoglycan degradation property of B. bifidum which is mediated by the complementary activities of SiaBb1 and SiaBb2 in the context of sialic acid cross-feeding.

  27. A Bio-synthetic Hybrid Hydrogel Formed under Physiological Conditions Consisting of Mucin and a Synthetic Polymer Carrying Boronic Acid. 国際誌 査読有り

    Masaki Nakahata, Naoki Tominaga, Keishi Saito, Keita Nishiyama, Yuya Tanino, Kiyoshiro Saiki, Masaru Kojima, Shinji Sakai

    Macromolecular bioscience　22　(6)　e2200055　2022年6月

    DOI： 10.1002/mabi.202200055 　

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    Mucin-containing bio-synthetic hybrid hydrogel is successfully formed under physiological conditions upon mixing aqueous solutions of native mucin and synthetic polymers carrying boronic acids. The mechanical properties and stability of the hydrogel in physiological solutions, e.g., cell culture media, are tunable depending on the boronic acid content of polymers. The hydrogel dissolved in the physiological solutions releases native mucin and boronic acid-containing polymer, which can control the adhesion of mammalian cells to the surface.

  28. Vaginal mucus in mice: Developmental and gene expression features of epithelial mucous cells during pregnancy. 国際誌 査読有り

    Makoto Sugiyama, Nao Machida, Arata Yasunaga, Nanako Terai, Hanae Fukasawa, Hisaya K Ono, Ryosuke Kobayashi, Keita Nishiyama, Osamu Hashimoto, Shiro Kurusu, Kazuki Yoshioka

    Biology of reproduction　105　(5)　1272-1282　2021年8月20日

    DOI： 10.1093/biolre/ioab157 　

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    The vagina is the site of copulation and serves as the birth canal. It also provides protection against external pathogens. In mice, due to the absence of cervical glands, the vaginal epithelium is the main producer of vaginal mucus. The development and differentiation of vaginal epithelium-constituting cells and the molecular characteristics of vaginal mucus have not been thoroughly examined. Here, we characterized vaginal mucous cell development and the expression of mucus-related factors in pregnant mice. The vaginal mucous epithelium layer thickened and became multilayered after day 12 of pregnancy and secreted increasing amounts of mucus until early postpartum. Using histochemistry and transmission electron microscopy, we found supra-basal mucous cells as probable candidates for precursor cells. In vaginal mucous cells, the expression of TFF1, a stabilizer of mucus, was high, and some members of mucins and antimicrobial peptides (MUC5B and DEFB1) were expressed in a stage-dependent manner. In summary, this study presents the partial characterization of vaginal epithelial mucous cell lineage and expression of genes encoding several peptide substances that may affect vaginal tissue homeostasis and mucosal immunity during pregnancy and parturition.

  29. Roles of the Cell Surface Architecture of Bacteroides and Bifidobacterium in the Gut Colonization. 国際誌 査読有り

    Keita Nishiyama, Tatsunari Yokoi, Makoto Sugiyama, Ro Osawa, Takao Mukai, Nobuhiko Okada

    Frontiers in microbiology　12　754819-754819　2021年

    DOI： 10.3389/fmicb.2021.754819 　

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    There are numerous bacteria reside within the mammalian gastrointestinal tract. Among the intestinal bacteria, Akkermansia, Bacteroides, Bifidobacterium, and Ruminococcus closely interact with the intestinal mucus layer and are, therefore, known as mucosal bacteria. Mucosal bacteria use host or dietary glycans for colonization via adhesion, allowing access to the carbon source that the host's nutrients provide. Cell wall or membrane proteins, polysaccharides, and extracellular vesicles facilitate these mucosal bacteria-host interactions. Recent studies revealed that the physiological properties of Bacteroides and Bifidobacterium significantly change in the presence of co-existing symbiotic bacteria or markedly differ with the spatial distribution in the mucosal niche. These recently discovered strategic colonization processes are important for understanding the survival of bacteria in the gut. In this review, first, we introduce the experimental models used to study host-bacteria interactions, and then, we highlight the latest discoveries on the colonization properties of mucosal bacteria, focusing on the roles of the cell surface architecture regarding Bacteroides and Bifidobacterium.

  30. Extracellular vesicles produced by Bifidobacterium longum export mucin-binding proteins. 国際誌 査読有り

    Keita Nishiyama, Takashi Takaki, Makoto Sugiyama, Itsuko Fukuda, Maho Aiso, Takao Mukai, Toshitaka Odamaki, Jin-Zhong Xiao, Ro Osawa, Nobuhiko Okada

    Applied and environmental microbiology　86　(19)　2020年7月31日

    DOI： 10.1128/AEM.01464-20 　

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    Extracellular proteins are important factors in host-microbe interactions; however, the specific factors that enable bifidobacterial adhesion and survival in the gastrointestinal (GI) tract are not fully characterized. Here, we discovered that Bifidobacterium longum NCC2705 cultured in bacteria-free supernatants of human fecal fermentation broth released a myriad of particles into the extracellular environment. The aim of this study was to characterize the physiological properties of these extracellular particles. The particles were approximately 50-80 nm in diameter with high protein and dsDNA content, suggesting that they were extracellular vesicles (EVs). A proteomics analysis showed that the EVs primarily consisted of cytoplasmic proteins with crucial functions in essential cellular processes. We identified several mucin-binding proteins using biomolecular interaction analysis, including phosphoketolase, GroEL, EF-Tu, phosphoglycerate kinase, transaldolase (Tal), and Hsp20. The recombinant GroEL and Tal proteins showed high binding affinities to mucin. Further, the immobilization of these proteins on microbeads affected the permanence of microbeads in the murine GI tract. These results suggest that bifidobacterial exposure conditions that mimic the intestine stimulate B. longum EV production. The resulting EVs exported several cytoplasmic proteins that may have promoted B. longum adhesion. This study improved our understanding of the Bifidobacterium colonization strategy in the intestinal microbiome.ImportanceBifidobacterium is a natural inhabitant of the human gastrointestinal (GI) tract. Morphological observations revealed that extracellular appendages of bifidobacteria in complex microbial communities are important for understanding its adaptations to the GI tract environment. We identified dynamic extracellular vesicle (EV) production by Bifidobacterium longum in bacteria-free fecal fermentation broth that was strongly suggestive of differing bifidobacterial extracellular appendages in the GI tract. In addition, export of the adhesive moonlighting proteins mediated by EVs may promote bifidobacterial colonization. This study provides new insight into the roles of EVs on bifidobacterial colonization processes as they adapt to the GI environment.

  31. Lactobacillus plantarum由来菌体表層グリセルアルデヒド-3-リン酸脱水素酵素の結晶構造解析:水銀結合メカニズムの解明 査読有り

    米田一成, 緒方美月, 西山啓太, 福田健二, 安田伸, 井越敬司, 木下英樹

    ミルクサイエンス(Web)　68　(1)　3‐11(J‐STAGE)-11　2019年4月

    出版者・発行元： 日本酪農科学会

    ISSN：2188-0700

  32. A new approach for analyzing an adhesive bacterial protein in the mouse gastrointestinal tract using optical tissue clearing. 国際誌 査読有り

    Keita Nishiyama, Makoto Sugiyama, Hiroki Yamada, Kyoko Makino, Sayaka Ishihara, Takashi Takaki, Takao Mukai, Nobuhiko Okada

    Scientific reports　9　(1)　4731-4731　2019年3月18日

    DOI： 10.1038/s41598-019-41151-y 　

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    Several bacterial moonlighting proteins act as adhesion factors, which are important for bacterial colonization of the gastrointestinal (GI) tract. However, little is known about the adherence properties of moonlighting proteins in the GI tract. Here, we describe a new approach for visualizing the localization of moonlighting protein-coated fluorescent microbeads in the whole GI tract by using a tissue optical clearing method, using elongation factor Tu (EF-Tu) as an example. As a bacterial cell surface-localized protein mimic, recombinant EF-Tu from Lactobacillus reuteri was immobilized on microbeads. EF-Tu-coating promoted the interaction of the microbeads with a Caco-2 cell monolayer. Next, the microbeads were orally administered to mice. GI whole tissues were cleared in aqueous fructose solutions of increasing concentrations. At 1 h after administration, the microbeads were diffused from the stomach up to the cecum, and after 3 h, they were diffused throughout the intestinal tract. In the lower digestive tract, EF-Tu-beads were significantly more abundant than non-coated control beads, suggesting that EF-Tu plays an important role in the persistence of the microbeads in the GI tract. The new approach will help in evaluating how moonlighting proteins mediate bacterial colonization.

  33. Adhesion of Lactobacillus to Intestinal Mucin. 国際誌

    Keita Nishiyama, Takao Mukai

    Methods in molecular biology (Clifton, N.J.)　1887　159-166　2019年

    DOI： 10.1007/978-1-4939-8907-2_14 　

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    Adhesion to intestinal mucin is one of the main in vitro tests for the study of probiotic strains. Mucins immobilized on microtiter plates and glass slides are easy and excellent methods used for the quantitative analysis of Lactobacillus adhesion. However, to maintain the performance of the quantitative analysis, these methods must be chosen appropriately according to the nature and characteristics of the strain, such as tolerance to surfactant and the ability to self-aggregate. Here we describe two methodologies used to evaluate adhesion of Lactobacillus to mucin, in addition to the isolation and purification of mucins from porcine colonic tissues.

  34. Adhesion of Lactobacillus to Intestinal Mucin. 国際誌 査読有り

    Nishiyama K, Mukai T

    Methods in molecular biology (Clifton, N.J.)　1887　159-166　2019年

    DOI： 10.1007/978-1-4939-8907-2_14 　

    ISSN：1064-3745

  35. Two extracellular sialidases from Bifidobacterium bifidum promote the degradation of sialyl-oligosaccharides and support the growth of Bifidobacterium breve. 国際誌 査読有り

    Nishiyama K, Nagai A, Uribayashi K, Yamamoto Y, Mukai T, Okada N

    Anaerobe　52　22-28　2018年8月

    DOI： 10.1016/j.anaerobe.2018.05.007 　

    ISSN：1075-9964

  36. Bifidobacterium bifidum Extracellular Sialidase Enhances Adhesion to the Mucosal Surface and Supports Carbohydrate Assimilation 国際誌 査読有り

    Keita Nishiyama, Yuji Yamamoto, Makoto Sugiyama, Takashi Takaki, Tadasu Urashima, Satoru Fukiya, Atsushi Yokota, Nobuhiko Okada, Takao Mukai

    MBIO　8　(5)　2017年9月

    DOI： 10.1128/mBio.00928-17 　

    ISSN：2150-7511

  37. Anchorless cell surface proteins function as laminin-binding adhesins in Lactobacillus rhamnosus FSMM22 国際誌 査読有り

    Ni Putu Desy Aryantini, Daisuke Kondoh, Keita Nishiyama, Yuji Yamamoto, Takao Mukai, I. Nengah Sujaya, Tadasu Urashima, Kenji Fukuda

    FEMS MICROBIOLOGY LETTERS　364　(6)　2017年3月

    DOI： 10.1093/femsle/fnx056 　

    ISSN：0378-1097

    eISSN：1574-6968

  38. Adhesion Properties of Lactic Acid Bacteria on Intestinal Mucin. 国際誌 査読有り

    Nishiyama K, Sugiyama M, Mukai T

    Microorganisms　4　(3)　2016年9月

    DOI： 10.3390/microorganisms4030034 　

  39. Lactobacillus rhamnosus GG SpaC pilin subunit binds to the carbohydrate moieties of intestinal glycoconjugates 国際誌 査読有り

    Keita Nishiyama, Shintaro Ueno, Makoto Sugiyama, Yuji Yamamoto, Takao Mukai

    ANIMAL SCIENCE JOURNAL　87　(6)　809-815　2016年6月

    DOI： 10.1111/asj.12491 　

    ISSN：1344-3941

    eISSN：1740-0929

  40. Adhesion properties of a putative polymorphic fimbrial subunit protein from Bifidobacterium longum subsp. longum. 査読有り

    Suzuki K, Nishiyama K, Miyajima H, Osawa R, Yamamoto Y, Mukai T

    Bioscience of microbiota, food and health　35　(1)　19-27　2016年

    DOI： 10.12938/bmfh.2015-015 　

    ISSN：2186-6953

  41. Cell surface-associated aggregation-promoting factor from Lactobacillus gasseri SBT2055 facilitates host colonization and competitive exclusion of Campylobacter jejuni. 国際誌 査読有り

    Nishiyama K, Nakazato A, Ueno S, Seto Y, Kakuda T, Takai S, Yamamoto Y, Mukai T

    Molecular microbiology　98　(4)　712-726　2015年11月

    DOI： 10.1111/mmi.13153 　

    ISSN：0950-382X

    eISSN：1365-2958

  42. Mouse Models for Assessing the Protective Efficacy of Lactobacillus gasseri SBT2055 against Helicobacter suis Infection Associated with the Development of Gastric Mucosa-Associated Lymphoid Tissue Lymphoma 国際誌 査読有り

    Hidenori Matsui, Tetsufumi Takahashi, Anders Overby, Somay Yamagata Murayama, Haruno Yoshida, Yuji Yamamoto, Keita Nishiyama, Yasuyuki Seto, Takashi Takahashi, Takao Mukai, Masahiko Nakamura

    HELICOBACTER　20　(4)　291-298　2015年8月

    DOI： 10.1111/hel.12203 　

    ISSN：1083-4389

    eISSN：1523-5378

  43. Adhesion properties of Lactobacillus rhamnosus mucus-binding factor to mucin and extracellular matrix proteins. 国際誌 査読有り

    Keita Nishiyama, Koichi Nakamata, Shintaro Ueno, Akari Terao, Ni Putu Desy Aryantini, I Nengah Sujaya, Kenji Fukuda, Tadasu Urashima, Yuji Yamamoto, Takao Mukai

    Bioscience, biotechnology, and biochemistry　79　(2)　271-9　2015年

    DOI： 10.1080/09168451.2014.972325 　

    ISSN：0916-8451

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    We previously described potential probiotic Lactobacillus rhamnosus strains, isolated from fermented mare milk produced in Sumbawa Island, Indonesia, which showed high adhesion to porcine colonic mucin (PCM) and extracellular matrix (ECM) proteins. Recently, mucus-binding factor (MBF) was found in the GG strain of L. rhamnosus as a mucin-binding protein. In this study, we assessed the ability of recombinant MBF protein from the FSMM22 strain, one of the isolates of L. rhamnosus from fermented Sumbawa mare milk, to adhere to PCM and ECM proteins by overlay dot blot and Biacore assays. MBF bound to PCM, laminin, collagen IV, and fibronectin with submicromolar dissociation constants. Adhesion of the FSMM22 mbf mutant strain to PCM and ECM proteins was significantly less than that of the wild-type strain. Collectively, these results suggested that MBF contribute to L. rhamnosus host colonization via mucin and ECM protein binding.

  44. Lactobacillus gasseri SBT2055 Reduces Infection by and Colonization of Campylobacter jejuni 国際誌 査読有り

    Keita Nishiyama, Yasuyuki Seto, Kazuki Yoshioka, Tsutomu Kakuda, Shinji Takai, Yuji Yamamoto, Takao Mukai

    PLOS ONE　9　(9)　e108827　2014年9月

    DOI： 10.1371/journal.pone.0108827 　

    ISSN：1932-6203

  45. Evaluation of bifidobacterial adhesion to acidic sugar chains of porcine colonic mucins 国際誌 査読有り

    Keita Nishiyama, Akira Kawanabe, Hirofumi Miyauchi, Fumiaki Abe, Daigo Tsubokawa, Kazuhiko Ishihara, Yuji Yamamoto, Takao Mukai

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY　78　(8)　1444-1451　2014年8月

    DOI： 10.1080/09168451.2014.918491 　

    ISSN：0916-8451

    eISSN：1347-6947

  46. Identification and Characterization of Sulfated Carbohydrate-Binding Protein from Lactobacillus reuteri 査読有り

    Keita Nishiyama, Ayaka Ochiai, Daigo Tsubokawa, Kazuhiko Ishihara, Yuji Yamamoto, Takao Mukai

    PLOS ONE　12　(3)　e0174257　2013年3月

    DOI： 10.1371/journal.pone.0174257 　

    ISSN：1932-6203

  47. Identification and characterization of sulfated carbohydrate-binding protein from Lactobacillus reuteri. 国際誌 査読有り

    Keita Nishiyama, Ayaka Ochiai, Daigo Tsubokawa, Kazuhiko Ishihara, Yuji Yamamoto, Takao Mukai

    PloS one　8　(12)　e83703　2013年

    DOI： 10.1371/journal.pone.0083703 　

    詳細を見る 詳細を閉じる

    We previously purified a putative sulfated-galactosylceramide (sulfatide)-binding protein with a molecular weight of 47 kDa from the cell surface of Lactobacillus reuteri JCM1081. The aim of this study was to identify the 47-kDa protein, examine its binding to sulfated glycolipids and mucins, and evaluate its role in bacterial adhesion to mucosal surfaces. By cloning and sequencing analysis, the 47-kDa protein was identified as elongation factor-Tu (EF-Tu). Adhesion properties were examined using 6 × Histidine-fused EF-Tu (His6-EF-Tu). Surface plasmon resonance analysis demonstrated pH-dependent binding of His6-EF-Tu to sulfated glycolipids, but not to neutral or sialylated glycolipids, suggesting that a sulfated galactose residue was responsible for EF-Tu binding. Furthermore, His6-EF-Tu was found to bind to porcine gastric mucin (PGM) by enzyme-linked immunosorbent assay. Binding was markedly reduced by sulfatase treatment of PGM and in the presence of acidic and desialylated oligosaccharide fractions containing sulfated carbohydrate residues prepared from PGM, demonstrating that sulfated carbohydrate moieties mediated binding. Histochemical staining revealed similar localization of His6-EF-Tu and high iron diamine staining in porcine mucosa. These results indicated that EF-Tu bound PGM via sulfated carbohydrate moieties. To characterize the contribution of EF-Tu to the interaction between bacterial cells and PGM, we tested whether anti-EF-Tu antibodies could inhibit the interaction. Binding of L. reuteri JCM1081 to PGM was significantly blocked in a concentration-dependent matter, demonstrating the involvement of EF-Tu in bacterial adhesion. In conclusion, the present results demonstrated, for the first time, that EF-Tu bound sulfated carbohydrate moieties of sulfated glycolipids and sulfomucin, thereby promoting adhesion of L. reuteri to mucosal surfaces.

  48. Isolation of Potential Probiotic Lactobacillus rhamnosus Strains from Traditional Fermented Mare Milk Produced in Sumbawa Island of Indonesia 査読有り

    Tala Shi, Keita Nishiyama, Koichi Nakamata, Ni Putu Desy Aryantini, Dai Mikumo, Yuji Oda, Yuji Yamamoto, Takao Mukai, I. Nengah Sujaya, Tadasu Urashima, Kenji Fukuda

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY　76　(10)　1897-1903　2012年10月

    DOI： 10.1271/bbb.120385 　

    ISSN：0916-8451

    eISSN：1347-6947

︎全件表示 ︎最初の5件までを表示

MISC 46

  1. Production of bioactive compounds through Food by-product metabolism by pig gut microbiota

    豊吉 瑛太, 生井 楓, 渡邊 真広, 吉田 安佑, 石原 沙耶花, 小田巻 俊, 大坪 和香, 前川 正充, 西山 啓太, 北澤 春樹

    日本畜産学会第 132 回大会　2024年8月

  2. 腸-肺軸を介する「ポストイムノバイオティクス」の抗ウイルス免疫機能評価

    福山皓太朗, 福山皓太朗, 生井楓, 生井楓, 大坪和香子, 大坪和香子, 西山啓太, VILLENA Julio, VILLENA Julio, 北澤春樹, 北澤春樹

    日本畜産学会大会講演要旨　132nd　2024年

    ISSN： 1342-4688

  3. 乳酸菌によるシアロオリゴ糖代謝から見た子豚の腸内細菌叢形成

    橋本凌河, 西山啓太, 生井楓, 鈴木香澄, 鈴木香澄, 福田伊津子, 杉山友太, 大坪和香子, 吉岡豪, 中崎瑛里, 北澤春樹

    ミルクサイエンス(Web)　73　(2)　2024年

    ISSN： 2188-0700

  4. 腸内細菌の共生と消化管定着

    西山啓太

    関西大学先端科学技術シンポジウム講演集　28th　2024年

  5. 乳酸菌によるシアロオリゴ糖代謝に着目した子豚の腸内細菌叢形成とその意義

    橋本凌河, 西山啓太, 生井楓, 鈴木香澄, 鈴木香澄, 福田伊津子, 杉山友太, 豊吉瑛太, 佐久間大河, 大坪和香子, 吉岡豪, 中崎瑛里, 北澤春樹

    日本乳酸菌学会誌　35　(2)　2024年

    ISSN： 1343-327X

  6. 腸内細菌との共生から見えてきたビフィズス菌の消化管定着機構

    西山啓太, 向井孝夫

    アグリバイオ　6　(14)　2023年

    ISSN： 2432-5511

  7. 細菌の共生関係から紐解く腸内細菌の消化管定着

    西山啓太

    腸内細菌学雑誌　37　(2)　2023年

    ISSN： 1343-0882

  8. 腸内細菌が産生する芳香族アミノ酸による腸管病原菌の感染制御

    西山啓太

    Institute for Fermentation, Osaka. Research Communications　(37)　2023年

    ISSN： 0073-8751

  9. 新規抗ウイルス性「ポストイムノバイオティクス」の選抜・機構解明

    今村圭哉, 今村圭哉, 荒木祥多, 荒木祥多, 生井楓, 生井楓, 宮崎綾子, 須田遊人, 大坪和香子, 大坪和香子, 西山啓太, 西山啓太, 北澤春樹, 北澤春樹

    日本畜産学会大会講演要旨　131st　2023年

    ISSN： 1342-4688

  10. ブタ腸管上皮細胞においてミトコンドリア機能を改善するポストイムノバイオティクスの探索

    生井楓, 生井楓, 鈴木眞依, 笠原日菜子, 西山啓太, 西山啓太, 大坪和香子, 大坪和香子, 北澤春樹, 北澤春樹

    日本畜産学会大会講演要旨　131st　2023年

    ISSN： 1342-4688

  11. 乳腺上皮細胞における乳成分の炎症制御に関する基礎的研究

    福山皓太朗, 生井楓, 生井楓, 周冰卉, 周冰卉, 大坪和香子, 大坪和香子, 西山啓太, 西山啓太, 麻生久, 麻生久, 北澤春樹, 北澤春樹

    ミルクサイエンス(Web)　72　(2)　2023年

    ISSN： 2188-0700

  12. シアル酸がもたらす腸内細菌叢への影響

    西山啓太

    ミルクサイエンス(Web)　71　(1)　2022年

    ISSN： 2188-0700

  13. ハマダラカの腸内細菌Methylobacterium sp.が卵発育に与える影響

    箱崎純, 野々垣雄介, 田邊太志, 西山啓太, 中山和彦, 原口麻子, 中村咲蓮, 草木迫浩大, 筏井宏実

    衛生動物　73　(2)　2022年

    ISSN： 0424-7086

  14. ウマ腸内細菌Streptococcus equinusによるイヌリンとセロビオースの菌株依存的資化性

    大坪和香子, 西村順子, 神山結斗, 周冰卉, 西山啓太, 北澤春樹

    日本ウマ科学会学術集会講演要旨集　35th　2022年

  15. 糖鎖を介した腸内細菌と宿主との相互作用

    西山啓太

    ミルクサイエンス(Web)　70　(2)　2021年

    ISSN： 2188-0700

  16. 腸内細菌の消化管定着

    西山啓太

    Journal of Environmental Biotechnology (Web)　20　(1)　2020年

    ISSN： 2436-5041

  17. 乳酸菌の機能と活用 乳酸菌の消化管定着

    西山啓太

    Bio Industry　37　(5)　2020年

    ISSN： 0910-6545

  18. イムノバイオティックLactobacillus rhamnosusのMBFノックアウト株の樹立

    友常 加惠, 水野 滉也, 周 冰卉, Albarracin Leonardo, Islam Md Aminul, 西山 啓太, 大坪 和香子, Villena Julio, 佐々木 泰子, 北澤 春樹

    ミルクサイエンス　68　(2)　148-148　2019年8月

    出版者・発行元： 日本酪農科学会

    ISSN： 1343-0289

    eISSN： 2188-0700

  19. 組織透明化から紐解く腸内細菌の定着プロセス 招待有り

    西山啓太, 杉山真言

    academist Journal　2019年7月

  20. イムノバイオティックLactobacillus rhamnosusのMBFノックアウト株の樹立

    友常加惠, 友常加惠, 友常加惠, 水野滉也, 水野滉也, 水野滉也, 周冰卉, 周冰卉, 周冰卉, ALBARRACIN Leonardo, ISLAM Md Aminul, ISLAM Md Aminul, ISLAM Md Aminul, 西山啓太, 大坪和香子, 大坪和香子, 大坪和香子, VILLENA Julio, 佐々木泰子, 北澤春樹, 北澤春樹, 北澤春樹

    ミルクサイエンス(Web)　68　(2)　2019年

    ISSN： 2188-0700

  21. 腸内細菌の宿主消化管における定着戦略

    西山啓太

    環境バイオテクノロジー学会大会プログラム講演要旨集　2019　2019年

  22. 腸内細菌の消化管ムチンとの相互作用を介した定着機構

    西山啓太

    日本食品科学工学会大会講演集　66th　2019年

    ISSN： 2759-3843

  23. シアル酸をめぐる腸内での競争

    西山啓太

    生物工学会誌　96　(10)　595　2018年10月25日

    ISSN： 0919-3758

  24. 消化管ムチンとの相互作用を介したビフィズス菌の定着戦略

    西山啓太, 岡田信彦

    バイオサイエンスとインダストリー　76　(4)　288‐291　2018年7月10日

    ISSN： 0914-8981

  25. 組織透明化法を用いたムーンライティングタンパク質の腸管付着因子としての機能解析

    西山啓太, 杉山真言, 向井孝夫, 岡田信彦

    日本乳酸菌学会誌　29　(2)　110　2018年7月2日

    ISSN： 1343-327X

  26. Bifidobacterium bifidumの細胞外シアリダーゼの腸粘液との相互作用と糖の資化における役割

    西山啓太, 長井暁, 吹谷智, 横田篤, 山本裕司, 向井孝夫, 岡田信彦

    腸内細菌学雑誌　32　(2)　102　2018年4月

    ISSN： 1343-0882

  27. ビフィズス菌と宿主腸粘膜との相互作用に関わる因子

    西山啓太, 向井孝夫

    日本乳酸菌学会誌　29　(1)　13‐18-18　2018年3月10日

    出版者・発行元： 日本乳酸菌学会

    ISSN： 1343-327X

  28. ビフィズス菌の宿主腸粘膜への定着に関わる菌体表層タンパク質の機能解析

    西山啓太

    日本乳酸菌学会誌　28　(3)　184‐185　2017年11月10日

    ISSN： 1343-327X

  29. Bifidobacterium longum subsp.longumが有するFim線毛の腸管定着における役割とポリマー化に関わるモチーフの探索

    西山啓太, 菅原宏祐, 小田巻俊孝, 福田真嗣, 高木孝士, 山本裕司, 向井孝夫, 清水(肖)金忠, 岡田信彦, 大澤朗

    日本乳酸菌学会誌　28　(2)　129　2017年6月27日

    ISSN： 1343-327X

  30. Bifidobacterium bifidumにおけるシアリダーゼは宿主との相互作用を促進し糖の資化をサポートする

    西山啓太, 山本裕司, 杉山真言, 篠原有香, 浦島匡, 吹谷智, 横田篤, 岡田信彦, 向井孝夫

    日本農芸化学会大会講演要旨集(Web)　2017　ROMBUNNO.3C18p05 (WEB ONLY)　2017年3月5日

    ISSN： 2186-7976

  31. Bifidobacterium longum subsp.longumの有するBL0675線毛の多型がムチンへの付着性に及ぼす影響

    鏡谷竹生, 西山啓太, 大澤朗, 山本裕司, 向井孝夫

    ミルクサイエンス　66　(2)　2017年

    ISSN： 1343-0289

  32. The elongation factor Tu from Lactobacillus reuteri inhibits the adhesion of Helicobacter pylori to porcine gastric mucin

    西山啓太, 鏡谷竹生, 山本裕司, 岡田信彦, 向井孝夫

    ミルクサイエンス　66　(1)　17‐26(J‐STAGE)-26　2017年

    出版者・発行元： 日本酪農科学会

    DOI： 10.11465/milk.66.17 　

    ISSN： 1343-0289

    詳細を見る 詳細を閉じる

    <p> <i>Helicobacter pylori</i>は，ヒトの胃に感染するグラム陰性のラセン状細菌であり，世界人口の約半数が感染している。プロバイオティクスには，宿主腸粘膜における病原細菌との付着部位の競合を介した感染阻害効果が知られている。筆者らは， <i>Lactobacillus reuteri</i> JCM1081の菌体表層由来Elongation factor Tu (EF-Tu) が<i>H. pylori</i>の腸粘膜におけるレセプターの一つである硫酸化糖鎖に結合することを見出してきた。本論文では，組み換えEF-Tuタンパク質 (His-EF-Tu) を用いて <i>H. pylori</i>の付着阻害の評価を目的とした。His-EF-Tuの添加は， <i>H. pylori</i>のブタ胃ムチン (PGM) への付着を用量依存的に阻害した。また， <i>H. pylori</i>臨床分離株においても同様の阻害効果が確認された。興味深いことに，PGMのスルファターゼ処理も同様に<i>H. pylori</i>の付着を減少させたが，一方で，スルファターゼ処理を行うとHis-EF-Tuによる阻害効果は，ほとんど確認されなかった。さらに，His-EF-Tuによる阻害効果は，硫酸化糖鎖に対するPGM34モノクローナル抗体の処理と同程度であった。ウエスタンブロッティングにより，4菌株の<i>Lactobacillus</i>の菌体表層画分からEF-Tuが検出された。以上の結果から，我々は，EF-Tuが<i>H. pylori</i>のムチンへの付着を阻害することを証明すると共に，EF-Tuと<i>H. pylori</i>が同一の付着部位の競合を介して感染阻害が成立したと考えられた。</p>

  33. 乳酸菌及びビフィズス菌の消化管定着機構の解明と感染予防への応用

    西山啓太

    ミルクサイエンス　66　(3)　219‐226(J‐STAGE)-226　2017年

    出版者・発行元： 日本酪農科学会

    DOI： 10.11465/milk.66.219 　

    ISSN： 1343-0289

    詳細を見る 詳細を閉じる

    <p> <i>Lactobacillus</i>や<i>Bifidobacterium</i>は哺乳類の消化管に広く生息する共生細菌である。これらの細菌にとって宿主腸粘膜に対する付着性を有することは腸内環境での定着や生存を有利にすると考えられている。これまで筆者は，細菌と宿主腸粘膜の相互作用に着目し，<i>Lactobacillus</i>や<i>Bifidobacterium</i>において複数の付着因子を同定し特徴付けてきた。さらに見出された付着特性を利用し，病原細菌との付着部位の競合を利用した感染予防の有用性を証明してきた。本総説では，<i>Lactobacillus</i>による<i>Campylobacter</i>と<i>Helicobacter</i>の感染予防に関する取り組みとそのメカニズムについて論じるとともに，最近筆者らが見出した<i>Bifidobacterium</i>の糖質分解酵素を介した付着機構について紹介する。</p>

  34. Lactobacillus属やBifidobacterium属の宿主腸粘膜への付着性に関わるアドヘシンとその分子機構

    西山啓太, 向井孝夫

    日本乳酸菌学会誌　27　(3)　176‐186-186　2016年11月18日

    出版者・発行元： 日本乳酸菌学会

    DOI： 10.4109/jslab.27.176 　

    ISSN： 1343-327X

    詳細を見る 詳細を閉じる

    Lactobacillus 属や Bifidobacterium 属は、哺乳類の消化管に生息する共生細菌である。非運動性のこれらの細菌にとって、宿主腸粘膜への付着は、流動的な腸内環境で定着を有利にする生存戦略のひとつであると考えられる。近年、 Lactobacillus 属や Bifidobacterium 属の遺伝子ツールや解析技術が確かなものとなり、菌体表層のアドヘシンを介した宿主腸粘膜との相互作用が分子レベルで明らかにされてきた。本総説では、近年研究が進んでいる Sortase 依存性の表層タンパク質である Mucus-binding proteins（MucBPs）と線毛に着目し、これらを介した Lactobacillus 属や Bifidobacterium 属の腸粘膜への付着性に関わる分子機構について、我々の研究成果を交えながら述べたい。さらに、 Lactobacillus 属の付着特性を利用した病原細菌の感染予防に関する取り組みについても紹介したい。

  35. プロバイオティクスに関する最近の研究動向~乳酸菌やビフィズス菌のムチンとの相互作用~

    向井孝夫, 西山啓太

    乳業技術　65　23‐40　2016年7月1日

    ISSN： 1341-7878

  36. 乳酸菌によるカンピロバクターの感染予防とその作用機序の解明に向けた研究

    西山 啓太, 向井 孝夫

    畜産技術 = Livestock technology　(730)　2-6　2016年3月

    出版者・発行元： 畜産技術協会

    ISSN： 0389-1348

  37. 乳酸菌の腸粘膜への定着機構

    西山 啓太, 向井 孝夫

    化学と生物　54　(7)　471-477　2016年

    出版者・発行元： 公益社団法人 日本農芸化学会

    DOI： 10.1271/kagakutoseibutsu.54.471 　

    ISSN： 0453-073X

    詳細を見る 詳細を閉じる

    乳酸菌は，哺乳類の小腸から大腸に広く棲息するグラム陽性細菌である．乳酸菌を構成する最大の属である<i>Lactobacillus</i>属は，多岐にわたる有用効果が報告されており，近年では，民間伝承的な健康増進効果にとどまらず予防医学への応用も期待されている．一般に乳酸菌は積極的に摂取され宿主消化管で定着することが求められることから，複雑な腸内フローラを形成する消化管において，摂取された乳酸菌がどのようなプロセスを経て定着・共生することができるのか興味深い点である．本解説では，乳酸菌の生存戦略の一つである腸粘膜への付着に着目し，特にアドヘシン（付着因子）の細胞表層への提示機構とその役割について解説する．

  38. Lactobacillus 属や Bifidobacterium 属の宿主腸粘膜への付着性に関わるアドヘシンとその分子機構

    西山 啓太, 向井 孝夫

    日本乳酸菌学会誌　27　(3)　176-186　2016年

    出版者・発行元： 日本乳酸菌学会

    DOI： 10.4109/jslab.27.176 　

    ISSN： 1343-327X

    詳細を見る 詳細を閉じる

    <i>Lactobacillus </i>属や <i>Bifidobacterium </i>属は、哺乳類の消化管に生息する共生細菌である。非運動性のこれらの細菌にとって、宿主腸粘膜への付着は、流動的な腸内環境で定着を有利にする生存戦略のひとつであると考えられる。近年、 <i>Lactobacillus </i>属や <i>Bifidobacterium </i>属の遺伝子ツールや解析技術が確かなものとなり、菌体表層のアドヘシンを介した宿主腸粘膜との相互作用が分子レベルで明らかにされてきた。本総説では、近年研究が進んでいる Sortase 依存性の表層タンパク質である Mucus-binding proteins（MucBPs）と線毛に着目し、これらを介した <i>Lactobacillus </i>属や <i>Bifidobacterium </i>属の腸粘膜への付着性に関わる分子機構について、我々の研究成果を交えながら述べたい。さらに、 <i>Lactobacillus </i>属の付着特性を利用した病原細菌の感染予防に関する取り組みについても紹介したい。

  39. 一般講演 プロバイオティクスによる病原細菌の感染予防 (平成27年度酪農科学シンポジウム 21世紀ミルクサイエンスのブレークスルーをめざして)

    西山 啓太, 向井 孝夫

    ミルクサイエンス = Milk science　64　(3)　245-253　2015年12月

    出版者・発行元： 日本酪農科学会

    ISSN： 1343-0289

  40. ビフィズス菌におけるシアリダーゼのムチン付着因子としての新たな役割

    西山啓太, 西山啓太, 篠原有香, 浦島匡, 吹谷智, 横田篤, 山本裕司, 向井孝夫

    日本乳酸菌学会誌　26　(2)　2015年

    ISSN： 1343-327X

  41. 乳酸菌とムチンの相互作用 : 糖鎖を介した乳酸菌の腸粘膜への付着機構 (特集 乳に係わる糖鎖研究の最前線)

    西山 啓太, 向井 孝夫

    応用糖質科学 : 日本応用糖質科学会誌　5　(1)　38-43　2015年

    出版者・発行元： 日本応用糖質科学会

    ISSN： 2185-6427

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    乳酸菌は古くから世界中でヨーグルトなど様々な発酵食品の製造に用いられてきた我々の生活に深く結びついた細菌である。I.I. Mechnikovがヨーグルトによる不老長寿説を唱えて以来，乳酸菌の有用性について興味・関心が集まり，多くの研究者によって乳酸菌のもつ機能性が明らかにされてきた。すなわち，免疫賦活作用，感染防御作用，整腸作用など宿主に対して有用な効果を発揮し健康維持に役立つ乳酸菌はプロバイオティクスとして知られてきた。一方，乳酸菌の主な棲息域である十二指腸から小腸下部の腸粘膜には多数の粘液産生細胞が存在し，ムチン糖タンパク質を主成分とするムチン層に覆われている。ムチン層は，様々な物理的刺激や消化液，病原細菌の感染から腸粘膜を保護する役割に加え，腸内細菌の棲息の場として知られており，乳酸菌もムチンを消化管での定着の足掛かりとしていると考えられてきた。したがって，経口的に摂取された乳酸菌が流動的な腸内環境中でプロバイオティクス効果を持続的に発揮する要素の1つとしてムチンへの付着性が注目され，これまでにムチンに付着性を示すいくつかの乳酸菌とその付着機構が報告されてきた。また，ムチンとの相互作用を理解することは，乳酸菌のプロバイオティクスとしての利用のみならず腸内細菌としての生存戦略を考える上でも非常に興味に深い研究対象である。本稿では，乳酸菌とムチンの相互作用について最新の報告を取り入れながら，腸内環境中における乳酸菌の巧みな腸内定着機構について解説したい。

  42. プロバイオティクスに関する最近の研究動向 : 乳酸菌やビフィズス菌のムチンとの相互作用

    向井 孝夫, 西山 啓太

    乳業技術　65　23-40　2015年

    出版者・発行元： 日本乳業技術協会

    ISSN： 1341-7878

  43. ビフィズス菌由来シアリダーゼのムチン付着因子としての評価

    西山啓太, 吹谷智, 横田篤, 山本裕司, 向井孝夫

    日本畜産学会大会講演要旨　118th　2014年

    ISSN： 1342-4688

  44. Lactobacillus reuteriにおけるElongation factor Tuのシグナル配列非依存的な分泌機構に関する研究

    西山啓太, 山本裕司, 向井孝夫

    日本乳酸菌学会誌　25　(2)　2014年

    ISSN： 1343-327X

  45. ビフィズス菌のシアリダーゼは付着因子として機能する

    西山啓太, 吹谷智, 横田篤, 山本裕司, 向井孝夫

    ミルクサイエンス　63　(2)　2014年

    ISSN： 1343-0289

  46. シグナル配列非依存的に分泌される多機能タンパク質の役割と提示機構

    西山 啓太, 山本 裕司, 向井 孝夫

    日本乳酸菌学会誌 = Journal of Japan Society for Lactic Acid Bacteria　24　(2)　2013年6月20日

    ISSN： 1343-327X

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産業財産権 2

  1. 抗ヘリコバクター・ハイルマニー剤

    瀬戸 泰幸, 東 直樹, 向井 孝夫, 中村 正彦, 松井 英則, 山本 裕司, 西山 啓太

    産業財産権の種類: 特許権

  2. ビフィドバクテリウム属細菌の線毛形成誘導用の組成物

    西山啓太, 小山信裕, 大澤朗, 岡田信彦, 清水金忠, 向井孝夫, 供田洋

    産業財産権の種類: 特許権

共同研究・競争的資金等の研究課題 11

  1. 利他的ビフィズス菌のヒト糖鎖結合による腸内生存戦略を分子構造から理解する

    伏信 進矢, 片山 高嶺, 西山 啓太

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Scientific Research (A)

    研究機関：The University of Tokyo

    2023年4月1日 ～ 2028年3月31日

  2. 家畜健全育成向上戦略：イムノバイオティクスと自然免疫受容体の腸-肺軸シンフォニー

    北澤 春樹, 西山 啓太, 大坪 和香子, Binghui Zhou, 野地 智法, 竹之内 敬人, 新開 浩樹, 須田 義人

    2023年4月1日 ～ 2027年3月31日

  3. 細菌の定着性強化に向けた基盤技術創製ーバイオフィルム形成に学ぶー

    小川 哲弘, 西山 啓太, 田川 陽一, 角田 茂

    2023年4月1日 ～ 2026年3月31日

  4. 細菌の定着性強化に向けた基盤技術創製ーバイオフィルム形成に学ぶー

    小川 哲弘, 角田 茂, 田川 陽一, 西山 啓太

    2023年4月1日 ～ 2026年3月31日

  5. 腸内細菌を有効活用した家畜健全育成に貢献する機能性エコフィードの開発基盤創成

    西山 啓太, 北澤 春樹, 生井 楓

    2023年4月1日 ～ 2026年3月31日

  6. シアロオリゴ糖代謝を起点とした腸内細菌と宿主の共生機構の解明

    阪中 幹祥, 西山 啓太, 杉山 友太

    2023年4月1日 ～ 2026年3月31日

  7. ブタ小腸フローラ・免疫制御システムの統合評価系構築

    北澤 春樹, 西山 啓太, Binghui Zhou

    2023年6月30日 ～ 2025年3月31日

  8. 腸内細菌が産生する芳香族化合物による真菌の感染制御

    西山 啓太

    2022年4月1日 ～ 2025年3月31日

  9. なぜビフィズス菌は腸内に定着できるのか？- 腸内細菌との共生から紐解く生存戦略

    西山 啓太

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Early-Career Scientists

    研究機関：Keio University

    2020年4月1日 ～ 2022年3月31日

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    腸内細菌は芳香族アミノ酸を様々な物質に変換し，宿主の生理状態に多くの影響を及ぼすことが知られている。本研究では，BacteroidesとLactobacillusの協調的代謝から産生される芳香族アミノ酸の代謝産物AAAがBifidobacteriumの線毛の発現を誘導し腸内定着を促進することを見出した。本知見は，腸内細菌が産生する特定の代謝産物がBifidobacteriumの腸管定着を制御するシグナル分子として作用することを明らかにした。

  10. 乳酸菌におけるシグナル配列非依存的なタンパク質の分泌機構の解明

    西山 啓太

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Young Scientists (B)

    2017年4月1日 ～ 2020年3月31日

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    乳酸菌における既知の分泌シグナル配列を持たないタンパク質の分泌機構の解明を目的とする。細菌が分泌するタンパク質の中には，SecやTatといった既知の分泌シグナル配列のない細胞質内タンパク質が多数検出される。これらは，解糖系酵素やタンパク質合成といった細胞内で何らかの役割をもつ因子であるが，菌体外に分泌されることで，菌体表層タンパク質のような役割を果たす。例えば，腸粘膜への接着因子や細菌同士の凝集促進因子，さらには宿主細胞の免疫刺激因子としてなど，その機能は多岐にわたる。そのため，細胞内での本来の役割に加え，菌体外での複数の機能をもつ意味から，ムーンライティングタンパク質として注目されている。 本研究では，乳酸菌のムーンライティングタンパク質のひとつ，翻訳伸長因子（elongation Factor Tu, EF-Tu）を取り上げ，分泌機構に関する基礎的知見の取得を目指す。昨年度は，EF-Tuの分泌に寄与する領域の探索を主として実施し，特定のアミノ酸配列を同定することに成功した。そこで，本年度は(i) 推定膜輸送装置とEF-Tu分泌シグナルの相互作用について解析を実施するとともに，(ii) 膜小胞を介したEF-Tuの分泌の可能性についても検討することとした。

  11. ビフィズス菌の糖質分解酵素を介した新規定着メカニズムの解析

    西山 啓太, 岡田 信彦, 向井 孝夫, 山本 裕司, 横田 篤, 吹谷 智, 浦島 匡

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research Grant-in-Aid for Research Activity start-up

    研究種目：Grant-in-Aid for Research Activity start-up

    研究機関：Kitasato University

    2015年8月28日 ～ 2017年3月31日

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    ビフィズス菌は，ヒト消化管に生息する共生細菌である。本研究では，Bifidobacterium bifidumのシアリダーゼ（SiaBb2）に着目し，付着因子と栄養獲得因子の両面からその役割について解析した。本研究から，SiaBb2がHMOに含まれるシアル化糖鎖を切断することで効率的な糖の資化を可能にしていること，さらにSiaBb2がB. bifidumのムチンに対する付着性を促進することを見出した。これらは，シアリダーゼが腸管付着性と栄養獲得というビフィズス菌の生存に重要な役割を担う二機能性の因子であることを示しており，SiaBb2を介した新たな定着機構の存在を示すものと考えられる。

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