Updated on 2025/08/22

写真a

 
YOSHIDA Keitaro
 
Organization
School of Medicine Department of Microbiology Assistant Professor
Title
Assistant Professor
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Degree

  • 博士(農学) ( 2017.3   筑波大学 )

Research Interests

  • denitrification

  • wastewater treatment

  • phage

  • biofilm

  • lipase

  • Respiratory syncytial virus

  • 非運動性細菌

Research Areas

  • Life Science / Virology

  • Life Science / Applied microbiology

  • Life Science / Bacteriology

Professional Memberships

  • 日本細菌学会

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  • JAPAN SOCIETY FOR BIOSCIENCE, BIOTECHNOLOGY, AND AGROCHEMISTRY

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  • THE JAPANESE SOCIETY OF MICROBIAL ECOLOGY

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  • 日本ウイルス学会

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Papers

  • Bacillaceae serine proteases and Streptomyces epsilon-poly-l-lysine synergistically inactivate Caliciviridae by inhibiting RNA genome release Reviewed

    Soh Yamamoto, Noriko Ogasawara, Yuka Sudo-Yokoyama, Sachiko Sato, Nozomu Takata, Nana Yokota, Tomomi Nakano, Kyoko Hayashi, Akira Takasawa, Mayumi Endo, Masako Hinatsu, Keitaro Yoshida, Toyotaka Sato, Satoshi Takahashi, Kenichi Takano, Takashi Kojima, Jun Hiraki, Shin-ich Yokota

    Scientific Reports   14 ( 1 )   2024.7

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    Publishing type:Research paper (scientific journal)   Publisher:Springer Science and Business Media LLC  

    Abstract

    Human norovirus (HuNoV) is an enteric infectious pathogen belonging to the Caliciviridae family that causes occasional epidemics. Circulating alcohol-tolerant viral particles that are readily transmitted via food-borne routes significantly contribute to the global burden of HuNoV-induced gastroenteritis. Moreover, contact with enzymes secreted by other microorganisms in the environment can impact the infectivity of viruses. Hence, understanding the circulation dynamics of Caliciviridae is critical to mitigating epidemics. Accordingly, in this study, we screened whether environmentally abundant secretase components, particularly proteases, affect Caliciviridae infectivity. Results showed that combining Bacillaceae serine proteases with epsilon-poly-l-lysine (EPL) produced by Streptomyces—a natural antimicrobial—elicited anti-Caliciviridae properties, including against the epidemic HuNoV GII.4_Sydney_2012 strain. In vitro and in vivo biochemical and virological analyses revealed that EPL has two unique synergistic viral inactivation functions. First, it maintains an optimal pH to promote viral surface conformational changes to the protease-sensitive structure. Subsequently, it inhibits viral RNA genome release via partial protease digestion at the P2 and S domains in the VP1 capsid. This study provides new insights regarding the high-dimensional environmental interactions between bacteria and Caliciviridae, while promoting the development of protease-based anti-viral disinfectants.

    Other Link: https://www.nature.com/articles/s41598-024-65963-9

    DOI: 10.1038/s41598-024-65963-9

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  • Disruption of SMC-related genes promotes recombinant cholesterol esterase production in Burkholderia stabilis

    Kenji Konishi, Yoshiaki Yasutake, Shuji Muramatsu, Satomi Murata, Keitaro Yoshida, Koji Ishiya, Sachiyo Aburatani, Shin-ichi Sakasegawa, Tomohiro Tamura

    Applied Microbiology and Biotechnology   106 ( 24 )   8093 - 8110   2022.12

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    Publishing type:Research paper (scientific journal)   Publisher:Springer Science and Business Media LLC  

    Other Link: https://link.springer.com/article/10.1007/s00253-022-12277-3/fulltext.html

    DOI: 10.1007/s00253-022-12277-3

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  • Bacterial triacylglycerol lipase is a potential cholesterol esterase: Identification of a key determinant for sterol-binding specificity Reviewed International journal

    Yoshiaki Yasutake, Kenji Konishi, Shuji Muramatsu, Keitaro Yoshida, Sachiyo Aburatani, Shin-ichi Sakasegawa, Tomohiro Tamura

    International Journal of Biological Macromolecules   167   578 - 586   2021.1

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    Cholesterol esterase (Che) from Burkholderia stabilis (BsChe) is a homolog of well-characterized and industrially relevant bacterial triacylglycerol lipases (Lips). BsChe is a rare bacterial Lip enzyme that exhibits practical Che activity and is currently used in clinical applications to determine total serum cholesterol levels. To investigate the sterol specificity of BsChe, we determined the X-ray structure of BsChe. We discovered a local structural change in the active-site cleft, which might be related to substrate binding and product release. We also performed molecular docking studies by using the X-ray models of BsChe and cholesterol linoleate (CLL), the most favorable substrate for BsChe. The results showed that the sterol moieties of reasonable CLL docking poses localized to a specific active-site cleft surface formed by Leu266 and Ile287, which are unconserved among Burkholderia Lip homologs. Site-directed mutagenesis identified these residues as essential for the Che activity of BsChe, and Leu or Ile substitution conferred marked Che activity to Burkholderia Lips. In particular, Burkholderia cepacia and Burkholderia ubonensis Lips with the V266L/L287I double mutation exhibited ~50-fold and 500-fold higher Che activities than those of the wild-type enzymes, respectively. These results provide new insights into the substrate-binding mechanisms and selectivities of bacterial Lips.

    DOI: 10.1016/j.ijbiomac.2020.11.184

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  • Peculiarities of biofilm formation by Paracoccus denitrificans. Reviewed International journal

    Kana Morinaga, Keitaro Yoshida, Kohei Takahashi, Nobuhiko Nomura, Masanori Toyofuku

    Applied microbiology and biotechnology   2020.1

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    Most bacteria form biofilms, which are thick multicellular communities covered in extracellular matrix. Biofilms can become thick enough to be even observed by the naked eye, and biofilm formation is a tightly regulated process. Paracoccus denitrificans is a non-motile, Gram-negative bacterium that forms a very thin, unique biofilm. A key factor in the biofilm formed by this bacterium is a large surface protein named biofilm-associated protein A (BapA), which was recently reported to be regulated by cyclic diguanosine monophosphate (cyclic-di-GMP or c-di-GMP). Cyclic-di-GMP is a major second messenger involved in biofilm formation in many bacteria. Though cyclic-di-GMP is generally reported as a positive regulatory factor in biofilm formation, it represses biofilm formation in P. denitrificans. Furthermore, quorum sensing (QS) represses biofilm formation in this bacterium, which is also reported as a positive regulator of biofilm formation in most bacteria. The QS signal used in P. denitrificans is hydrophobic and is delivered through membrane vesicles. Studies on QS show that P. denitrificans can potentially form a thick biofilm but maintains a thin biofilm under normal growth conditions. In this review, we discuss the peculiarities of biofilm formation by P. denitrificans with the aim of deepening the overall understanding of bacterial biofilm formation and functions.

    DOI: 10.1007/s00253-020-10400-w

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  • Production of recombinant extracellular cholesterol esterase using consistently active promoters in Burkholderia stabilis Reviewed International journal

    Keitaro Yoshida, Kenji Konishi, Arturo Magana-Mora, Adrien Rougny, Yoshiaki Yasutake, Shuji Muramatsu, Satomi Murata, Toshitaka Kumagai, Sachiyo Aburatani, Shin-ichi Sakasegawa, Tomohiro Tamura

    Bioscience, Biotechnology, and Biochemistry   83 ( 10 )   1974 - 1984   2019.10

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Oxford University Press (OUP)  

    <title>ABSTRACT</title>
    Burkholderia stabilis FERMP-21014 produces highly active cholesterol esterase in the presence of fatty acids. To develop an overexpression system for cholesterol esterase production, we carried out RNA sequencing analyses to screen strongly active promoters in FERMP-21014. Based on gene expression consistency analysis, we selected nine genes that were consistently expressed at high levels, following which we constructed expression vectors using their promoter sequences and achieved overproduction of extracellular cholesterol esterase under fatty acid-free conditions. Of the tested promoters, the promoter of BSFP_0720, which encodes the alkyl hydroperoxide reductase subunit AhpC, resulted in the highest cholesterol esterase activity (24.3 U mL−1). This activity level was 243-fold higher than that of the wild-type strain under fatty acid-free conditions. We confirmed that cholesterol esterase was secreted without excessive accumulation within the cells. The gene expression consistency analysis will be useful to screen promoters applicable to the overexpression of other industrially important enzymes.

    DOI: 10.1080/09168451.2019.1630256

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  • Genome Sequence of Rhodococcus erythropolis Type Strain JCM 3201 Reviewed International journal

    Keitaro Yoshida, Wataru Kitagawa, Koji Ishiya, Yasuo Mitani, Nobutaka Nakashima, Sachiyo Aburatani, Tomohiro Tamura

    Microbiology Resource Announcements   8 ( 14 )   2019.4

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:American Society for Microbiology  

    <named-content content-type="genus-species">Rhodococcus erythropolis</named-content>
    JCM 3201 can express several recombinant proteins that are difficult to express in
    <named-content content-type="genus-species">Escherichia coli</named-content>
    . It is used as one of the hosts for protein expression and bioconversion.

    DOI: 10.1128/mra.01730-18

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  • Complete Genome Sequence of an Efficient Vitamin D3-Hydroxylating Bacterium, Pseudonocardia autotrophica NBRC 12743 Reviewed

    Keitaro Yoshida, Yoshiaki Yasutake, Tomohiro Tamura

    Microbiology Resource Announcements   7 ( 12 )   2018.9

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    Authorship:Lead author   Publishing type:Research paper (scientific journal)   Publisher:American Society for Microbiology  

    <named-content content-type="genus-species">Pseudonocardia autotrophica</named-content>
    NBRC 12743 contains a cytochrome P450 vitamin D
    <sub>3</sub>
    hydroxylase, and it is used as a biocatalyst for the commercial production of hydroxyvitamin D
    <sub>3</sub>
    , a valuable compound for medication. Here, we report the complete genome sequence of
    <named-content content-type="genus-species">P. autotrophica</named-content>
    NBRC 12743, which could be useful for improving the productivity of hydroxyvitamin D
    <sub>3</sub>
    .

    DOI: 10.1128/mra.01105-18

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  • Nitric Oxide Production from Nitrite Reduction and Hydroxylamine Oxidation by Copper-containing Dissimilatory Nitrite Reductase (NirK) from the Aerobic Ammonia-oxidizing Archaeon, <i>Nitrososphaera viennensis</i> Reviewed

    Shun Kobayashi, Daisuke Hira, Keitaro Yoshida, Masanori Toyofuku, Yosuke Shida, Wataru Ogasawara, Takashi Yamaguchi, Nobuo Araki, Mamoru Oshiki

    Microbes and Environments   33 ( 4 )   428 - 434   2018

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    Publishing type:Research paper (scientific journal)   Publisher:Japanese Society of Microbial Ecology  

    DOI: 10.1264/jsme2.me18058

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  • Biofilm formation by Paracoccus denitrificans requires a type I secretion system-dependent adhesin BapA Reviewed

    Keitaro Yoshida, Masanori Toyofuku, Nozomu Obana, Nobuhiko Nomura

    FEMS Microbiology Letters   364 ( 4 )   2017

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1093/femsle/fnx029

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  • Impacts of hydrophilic colanic acid on bacterial attachment to microfiltration membranes and subsequent membrane biofouling Reviewed

    Keitaro Yoshida, Yosuke Tashiro, Thithiwat May, Satoshi Okabe

    Water Research   76   33 - 42   2015.6

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1016/j.watres.2015.02.045

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  • Nitrate-dependent ferrous iron oxidation by anaerobic ammonium oxidation (anammox) bacteria Reviewed

    M. Oshiki, S. Ishii, K. Yoshida, N. Fujii, M. Ishiguro, Hisashi Satoh, S. Okabe

    Applied and Environmental Microbiology   79 ( 13 )   4087 - 4093   2013.7

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

    Other Link: http://orcid.org/0000-0002-5017-8566

    DOI: 10.1128/AEM.00743-13

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MISC

  • 多剤耐性細菌感染症の切り札 ファージセラピー

    吉田圭太朗, 安藤弘樹

    臨床化学   51 ( 1 )   47 - 47   2022.1

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  • Crystal structure analysis of Burkholderia stabilis cholesterol esterase

    安武義晃, 安武義晃, 小西健司, 小西健司, 村松周治, 吉田圭太朗, 油谷幸代, 油谷幸代, 油谷幸代, 酒瀬川信一, 田村具博, 田村具博, 田村具博

    量子ビームサイエンスフェスタ(Web)   2020   2021

  • Production of recombinant extracellular cholesterol esterase using Burkholderia stabilis

    吉田圭太朗, 小西健司, MAGANA-MORA Arturo, MAGANA-MORA Arturo, ROUGNY Adrien, ROUGNY Adrien, 安武義晃, 安武義晃, 村松周治, 村田里美, 熊谷俊高, 油谷幸代, 油谷幸代, 酒瀬川信一, 田村具博, 田村具博, 田村具博

    日本農芸化学会大会講演要旨集(Web)   2020   2020

  • Burkholderia stabilis由来コレステロールエステラーゼの機能と臨床検査薬への適用

    小西健司, 吉田圭太朗, 太田喜之, 安武義晃, 村松周治, 村田里美, 油谷幸代, 酒瀬川信一, 田村具博

    日本農芸化学会西日本支部大会およびシンポジウム講演要旨集   2019   2019

  • Burkholderia stabilisのコレステロールエステラーゼはT2SS機構の内膜タンパク質GspFを介して分泌される

    吉田圭太朗, 小西健司, 村松周治, 酒瀬川信一, 安武義晃, 油谷幸代, 田村具博

    日本農芸化学会西日本支部大会およびシンポジウム講演要旨集   2019   2019

  • アンモニア酸化古細菌Nitrososphaera viennensisの亜硝酸還元酵素の異種発現および機能解析

    押木守, 小林駿, 荒木信夫, 平大輔, 吉田圭太朗, 豊福雅典, 志田洋介, 小笠原渉, 山口隆司

    日本水環境学会シンポジウム講演集   21st   181‐182   2018.9

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    Language:Japanese  

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  • データベースに眠る宝の山 シトクロムP450

    吉田圭太朗

    生物工学会誌   96 ( 8 )   473   2018.8

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  • Sociomicrobiology; microbial diversity in biofilm communities Invited

    Tomohiro Inaba, Nozomu Obana, Tatsunori Kiyokawa, Keitaro Yoshida, Fumihiro Sassa, Atsushi Ogata, Nobuhiko Nomura

    177 ( 3 )   107 - 129   2015.12

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    Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (other)  

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  • 嫌気性アンモニウム酸化細菌のnirS遺伝子の機能解析

    小林駿, 押木守, 蝶名林郁也, 荒木信夫, 吉田圭太郎, 豊福雅典

    日本微生物生態学会大会(Web)   30th   PD‐061(JTK) (WEB ONLY)   2015

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    Language:Japanese  

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  • 3P-159 Analysis of adhesins of a non-motlile Gram-negative bacterium Paracoccus denitrificans

    Yoshida Keitaro, Toyofuku Masanori, Nomura Nobuhiko

    67   310 - 310   2015

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Presentations

  • Recombinant production of extracellular cholesterol esterase using consistently active promoters in Burkholderia stabilis International conference

    Keitaro Yoshida

    2019 International Conference on Biotechnology and Bioengineering  2019.9 

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    Language:English   Presentation type:Oral presentation (general)  

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Research Projects

  • 複合微生物系における膜の目詰まりの新奇生物学的制御

    2014.4 - 2017.3

    日本学術振興会  科学研究費助成 特別研究員奨励費DC1 

    吉田圭太朗

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    Authorship:Principal investigator  Grant type:Competitive

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