Updated on 2024/03/15

写真a

 
YOSHIMOTO Shogo
 
Organization
Graduate School of Engineering Biomolecular Engineering 2 Assistant Professor
Graduate School
Graduate School of Engineering
Undergraduate School
School of Engineering Chemistry and Biotechnology
Title
Assistant Professor
External link

Degree 1

  1. Ph.D. ( 2017.3   Nagoya University ) 

Research Interests 5

  1. 微生物付着

  2. タンパク質工学

  3. 人工細胞

  4. 原子間力顕微鏡

  5. X線結晶構造解析

Research Areas 1

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

Current Research Project and SDGs 3

  1. 微生物を用いたものづくり

  2. 新しいタンパク質マテリアルの創出

  3. 酵素によるプラスチック分解

Education 1

  1. Nagoya University

    2012.4 - 2017.3

Professional Memberships 3

  1. 日本生物工学会

  2. 日本化学会

  3. 化学工学会

 

Papers 16

  1. Adhesion preference of the sticky bacterium Acinetobacter sp. Tol 5 Reviewed International coauthorship

    Shogo Yoshimoto, Satoshi Ishii, Ayane Kawashiri, Taishi Matsushita, Stephan Göttig, Volkhard Kempf, Madoka Takai, Katsutoshi Hori

    Frontiers in Bioengineering and Biotechnology   Vol. 12   page: 1342418   2024.2

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

    DOI: 10.3389/fbioe.2024.1342418

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  2. Simple Method for the Creation of a Bacteria-Sized Unilamellar Liposome with Different Proteins Localized to the Respective Sides of the Membrane Reviewed

    Noba Kosaku, Yoshimoto Shogo, Tanaka Yoshikazu, Yokoyama Takeshi, Matsuura Tomoaki, Hori Katsutoshi

    ACS Synthetic Biology   Vol. 12 ( 5 ) page: 1437 - 1446   2023.5

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACS Synthetic Biology  

    Artificial cells are membrane vesicles mimicking cellular functions. To date, giant unilamellar vesicles made from a single lipid membrane with a diameter of 10 μm or more have been used to create artificial cells. However, the creation of artificial cells that mimic the membrane structure and size of bacteria has been limited due to technical restrictions of conventional liposome preparation methods. Here, we created bacteria-sized large unilamellar vesicles (LUVs) with proteins localized asymmetrically to the lipid bilayer. Liposomes containing benzylguanine-modified phospholipids were prepared by combining the conventional water-in-oil emulsion method and the extruder method, and green fluorescent protein fused with SNAP-tag was localized to the inner leaflet of the lipid bilayer. Biotinylated lipid molecules were then inserted externally, and the outer leaflet was modified with streptavidin. The resulting liposomes had a size distribution in the range of 500-2000 nm with a peak at 841 nm (the coefficient of variation was 10.3%), which was similar to that of spherical bacterial cells. Fluorescence microscopy, quantitative evaluation using flow cytometry, and western blotting proved the intended localization of different proteins on the lipid membrane. Cryogenic electron microscopy and quantitative evaluation by α-hemolysin insertion revealed that most of the created liposomes were unilamellar. Our simple method for the preparation of bacteria-sized LUVs with asymmetrically localized proteins will contribute to the creation of artificial bacterial cells for investigating functions and the significance of their surface structure and size.

    DOI: 10.1021/acssynbio.2c00564

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  3. Identification of the adhesive domain of AtaA from Acinetobacter sp. Tol 5 and its application in immobilizing Escherichia coli Reviewed International coauthorship

    Yoshimoto Shogo, Aoki Sota, Ohara Yuki, Ishikawa Masahito, Suzuki Atsuo, Linke Dirk, Lupas Andrei N., Hori Katsutoshi

    FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY   Vol. 10   page: 1095057   2023.1

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    Authorship:Lead author   Language:English   Publisher:Frontiers in Bioengineering and Biotechnology  

    Cell immobilization is an important technique for efficiently utilizing whole-cell biocatalysts. We previously invented a method for bacterial cell immobilization using AtaA, a trimeric autotransporter adhesin from the highly sticky bacterium Acinetobacter sp. Tol 5. However, except for Acinetobacter species, only one bacterium has been successfully immobilized using AtaA. This is probably because the heterologous expression of large AtaA (1 MDa), that is a homotrimer of polypeptide chains composed of 3,630 amino acids, is difficult. In this study, we identified the adhesive domain of AtaA and constructed a miniaturized AtaA (mini-AtaA) to improve the heterologous expression of ataA. In-frame deletion mutants were used to perform functional mapping, revealing that the N-terminal head domain is essential for the adhesive feature of AtaA. The mini-AtaA, which contains a homotrimer of polypeptide chains from 775 amino acids and lacks the unnecessary part for its adhesion, was properly expressed in E. coli, and a larger amount of molecules was displayed on the cell surface than that of full-length AtaA (FL-AtaA). The immobilization ratio of E. coli cells expressing mini-AtaA on a polyurethane foam support was significantly higher compared to the cells with or without FL-AtaA expression, respectively. The expression of mini-AtaA in E. coli had little effect on the cell growth and the activity of another enzyme reflecting the production level, and the immobilized E. coli cells could be used for repetitive enzymatic reactions as a whole-cell catalyst.

    DOI: 10.3389/fbioe.2022.1095057

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  4. The extracellular juncture domains of Type 5 autotransporters

    Weikum Julia, Kulakova Alina, Tesei Giulio, Yoshimoto Shogo, Jaegerum Line Vejby, Schutz Monika, Hori Katsutoshi, Skepo Marie, Harris Pernille, Leo Jack C., Morth J. Preben

    ACTA CRYSTALLOGRAPHICA A-FOUNDATION AND ADVANCES   Vol. 78   page: E5 - E5   2022.8

  5. Recent advances in research on biointerfaces: From cell surfaces to artificial interfaces Reviewed

    Hori Katsutoshi, Yoshimoto Shogo, Yoshino Tomoko, Zako Tamotsu, Hirao Gen, Fujita Satoshi, Nakamura Chikashi, Yamagishi Ayana, Kamiya Noriho

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   Vol. 133 ( 3 ) page: 195 - 207   2022.3

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    Language:Japanese   Publisher:Journal of Bioscience and Bioengineering  

    Biointerfaces are regions where biomolecules, cells, and organic materials are exposed to environmental media or come in contact with other biomaterials, cells, and inorganic/organic materials. In this review article, six research topics on biointerfaces are described to show examples of state-of-art research approaches. First, biointerface design of nanoparticles for molecular detection is described. Functionalized gold nanoparticles can be used for sensitive detection of various target molecules, including chemical compounds and biomolecules, such as DNA, proteins, cells, and viruses. Second, the interaction between bacterial cell surfaces and material surfaces, including the introduction of advances in analytical methods and theoretical calculations, are explained as well as their applications to bioprocesses. Third, bioconjugation technologies for localizing functional proteins at biointerfaces are introduced, in particular, by focusing the potential of enzymes as a catalytic tool for designing different types of bioconjugates that function at biointerfaces. Forth topics is focusing on lipid–protein interaction in cell membranes as natural biointerfaces. Examples of membrane lipid engineering are introduced, and it is mentioned how their compositional profiles affect membrane protein functions. Fifth topic is the physical method for molecular delivery across the biointerface being developed currently, such as highly efficient nanoinjection, electroporation, and nanoneedle devices, in which the key is how to perforate the cell membrane. Final topic is the chemical design of lipid- or polymer-based RNA delivery carriers and their behavior on the cell interface, which are currently attracting attention as RNA vaccine technologies targeting COVID-19. Finally, future directions of biointerface studies are presented.

    DOI: 10.1016/j.jbiosc.2021.12.004

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  6. Single-cell adhesion force mapping of a highly sticky bacterium in liquid Reviewed

    Ishii Satoshi, Yoshimoto Shogo, Hori Katsutoshi

    JOURNAL OF COLLOID AND INTERFACE SCIENCE   Vol. 606 ( Pt 1 ) page: 628 - 634   2022.1

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    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:Journal of Colloid and Interface Science  

    The sticky bacterium Acinetobacter sp. Tol 5 adheres to various material surfaces via its cell surface nanofiber protein, AtaA. This adhesiveness has only been evaluated based on the amount of cells adhering to a surface. In this study, the adhesion force mapping of a single Tol 5 cell in liquid using the quantitative imaging mode of atomic force microscopy (AFM) revealed that the adhesion of Tol 5 was near 2 nN, which was 1–2 orders of magnitude higher than that of other adhesive bacteria. The adhesion force of a cell became stronger with the increase in AtaA molecules present on the cell surface. Many fibers of peritrichate AtaA molecules simultaneously interact with a surface, strongly attaching the cell to the surface. The adhesion force of a Tol 5 cell was drastically reduced in the presence of 1% casamino acids but not in deionized water (DW), although both liquids decrease the adhesiveness of Tol 5 cells, suggesting that DW and casamino acids inhibit the cell approaching step and the subsequent direct interaction step of AtaA with surfaces, respectively. Heterologous production of AtaA provided non-adhesive Acinetobacter baylyi ADP1 cells with a strong adhesion force to AFM tip surfaces of silicon and gold.

    DOI: 10.1016/j.jcis.2021.08.039

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  7. The extracellular juncture domains in the intimin passenger adopt a constitutively extended conformation inducing restraints to its sphere of action Reviewed International coauthorship

    Weikum Julia, Kulakova Alina, Tesei Giulio, Yoshimoto Shogo, Jaegerum Line Vejby, Schuetz Monika, Hori Katsutoshi, Skepo Marie, Harris Pernille, Leo Jack C., Morth J. Preben

    SCIENTIFIC REPORTS   Vol. 10 ( 1 ) page: 21249   2020.12

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    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:Scientific Reports  

    Enterohemorrhagic and enteropathogenic Escherichia coli are among the most important food-borne pathogens, posing a global health threat. The virulence factor intimin is essential for the attachment of pathogenic E. coli to the intestinal host cell. Intimin consists of four extracellular bacterial immunoglobulin-like (Big) domains, D00–D2, extending into the fifth lectin subdomain (D3) that binds to the Tir-receptor on the host cell. Here, we present the crystal structures of the elusive D00–D0 domains at 1.5 Å and D0–D1 at 1.8 Å resolution, which confirms that the passenger of intimin has five distinct domains. We describe that D00–D0 exhibits a higher degree of rigidity and D00 likely functions as a juncture domain at the outer membrane-extracellular medium interface. We conclude that D00 is a unique Big domain with a specific topology likely found in a broad range of other inverse autotransporters. The accumulated data allows us to model the complete passenger of intimin and propose functionality to the Big domains, D00–D0–D1, extending directly from the membrane.

    DOI: 10.1038/s41598-020-77706-7

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  8. Native display of a huge homotrimeric protein fiber on the cell surface after precise domain deletion Reviewed International coauthorship

    Aoki Sota, Yoshimoto Shogo, Ishikawa Masahito, Linke Dirk, Lupas Andrei, Hori Katsutoshi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   Vol. 129 ( 4 ) page: 412 - 417   2020.4

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    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:Journal of Bioscience and Bioengineering  

    AtaA, a trimeric autotransporter adhesin from Acinetobacter sp. Tol 5, exhibits nonspecific, high adhesiveness to abiotic surfaces. For identification of the functional domains of AtaA, precise design of domain-deletion mutants is necessary so as not to cause undesirable structural distortion. Here, we designed and constructed three types of AtaA mutants from which the same domain, FGG1, was deleted. The first mutant was designed to preserve the periodicity of hydrophobic residues in the coiled-coil segments sandwiching the deleted region. After the deletion, the protein was properly displayed on the cell surface and had the same adhesive function as the wild type. Transmission electron microscopy (TEM) imaging and circular dichroism (CD) spectroscopy showed that its isolated passenger domain had the same fiber structure as in the AtaA wild type. In contrast, a mutant designed to disturb the coiled-coil periodicity at the deletion site failed to reach the cell surface. Although secretion occurred for the mutant designed with a flexible connector between the coiled coils, the cells exhibited a decrease in adhesiveness. Furthermore, TEM imaging of the mutant fibers showed bending at the fiber tip and changes in their CD spectrum indicated a decrease in secondary structure content. Thus, we succeeded to natively display the huge homotrimeric fiber structure of AtaA on the cell surface after precise deletion of a domain, maintaining the proper folding state and adhesive function by preserving its coiled-coil periodicity. This strategy enables us to construct various domain-deletion mutants of AtaA without structural distortion for complete functional mapping.

    DOI: 10.1016/j.jbiosc.2019.09.022

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  9. Process Description of an Unconventional Biofilm Formation by Bacterial Cells Autoagglutinating through Sticky, Long, and Peritrichate Nanofibers Reviewed

    Furuichi Yoshihide, Yoshimoto Shogo, Inaba Tomohiro, Nomura Nobuhiko, Hori Katsutoshi

    ENVIRONMENTAL SCIENCE & TECHNOLOGY   Vol. 54 ( 4 ) page: 2520 - 2529   2020.2

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    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:Environmental Science and Technology  

    In this study, we elucidated the formation process of an unconventional biofilm formed by a bacterium autoagglutinating through sticky, long, and peritrichate nanofibers. Understanding the mechanisms of biofilm formation is essential to control microbial behavior and improve environmental biotechnologies. Acinetobacter sp. Tol 5 autoagglutinate through the interaction of the long, peritrichate nanofiber protein AtaA, a trimeric autotransporter adhesin. Using AtaA, without cell growth or extracellular polymeric substances production, Tol 5 cells quickly form an unconventional biofilm. The process forming this unconventional biofilm started with cell-cell interactions, proceeded to cell clumping, and led to the formation of large cell aggregates. The cell-cell interaction was described by Derjaguin-Landau-Verwey-Overbeek (DLVO) theory based on a new concept, which considers two independent interactions between two cell bodies and between two AtaA fiber tips forming a discontinuous surface. If cell bodies cannot collide owing to an energy barrier at low ionic strengths but approach within the interactive distance of AtaA fibers, cells can agglutinate through their contact. Cell clumping proceeds following the cluster-cluster aggregation model, and an unconventional biofilm containing void spaces and a fractal nature develops. Understanding its formation process would extend the utilization of various types of biofilms, enhancing environmental biotechnologies.

    DOI: 10.1021/acs.est.9b06577

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  10. Bottom-up Creation of an Artificial Cell Covered with the Adhesive Bacterionanofiber Protein AtaA Reviewed

    Noba Kosaku, Ishikawa Masahito, Uyeda Atsuko, Watanabe Takayoshi, Hohsaka Takahiro, Yoshimoto Shogo, Matsuura Tomoaki, Hori Katsutoshi

    JOURNAL OF THE AMERICAN CHEMICAL SOCIETY   Vol. 141 ( 48 ) page: 19058 - 19066   2019.12

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    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:Journal of the American Chemical Society  

    The bacterial cell surface structure has important roles for various cellular functions. However, research on reconstituting bacterial cell surface structures is limited. This study aimed to bottom-up create a cell-sized liposome covered with AtaA, the adhesive bacterionanofiber protein localized on the cell surface of Acinetobacter sp. Tol 5, without the use of the protein secretion and assembly machineries. Liposomes containing a benzylguanine derivative-modified phospholipid were decorated with a truncated AtaA protein fused to a SNAP-tag expressed in a soluble fraction in Escherichia coli. The obtained liposome showed a similar surface structure and function to that of native Tol 5 cells and adhered to both hydrophobic and hydrophilic solid surfaces. Furthermore, this artificial cell was able to drive an enzymatic reaction in the adhesive state. The developed artificial cellular system will allow for analysis of not only AtaA, but also other cell surface proteins under a cell-mimicking environment. In addition, AtaA-decorated artificial cells may inspire the development of biotechnological applications that require immobilization of cells onto a variety of solid surfaces, in particular, in environments where the use of genetically modified organisms is prohibited.

    DOI: 10.1021/jacs.9b09340

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  11. Control of AtaA-mediated bacterial immobilization by casein hydrolysates Reviewed

    Ohara Yuki, Yoshimoto Shogo, Hori Katsutoshi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   Vol. 128 ( 5 ) page: 544 - 550   2019.11

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    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:Journal of Bioscience and Bioengineering  

    Acinetobacter sp. Tol 5 exhibits an autoagglutinating nature and high adhesiveness to various abiotic surfaces through its bacterionanofiber protein AtaA. We have developed new bacterial immobilization methods utilizing the high adhesiveness of AtaA. We previously reported that salt is essential for the adhesiveness of AtaA. In the current study, we unexpectedly found that Tol 5 cells were not immobilized onto polyurethane foam support during growth in LB medium although AtaA was properly expressed and displayed onto the cell surface. The adhesion of Tol 5 resting cells was not affected by sugars but drastically inhibited by yeast extract and casein hydrolysates such as tryptone and casamino acids technical grade (CA-T). Some amino acids, which are major components of CA-T, partially inhibited the adhesion of Tol 5 cells. Experimental results suggested that oligopeptides might effectively inhibit the cell adhesion. Immobilized cells onto the support through AtaA were detached in CA-T solution. Also, the detached cells could be re-immobilized onto the support without impairing of their adhesiveness by replacing CA-T solution to a basal salt medium. Microscopic observation revealed that breaking of AtaA-mediated cell–cell interaction is important for the detachment of Tol 5 cells from the support. CA-T also inhibited AtaA-mediated autoagglutination and dispersed cell clumps through AtaA. This is the first report on adhesion inhibitors against AtaA and suggests that casein hydrolysates like CA-T would be a powerful tool for controlling AtaA-mediated bacterial immobilization.

    DOI: 10.1016/j.jbiosc.2019.04.019

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  12. 微生物の非特異的な細胞接着

    吉本 将悟

    生物工学会誌   Vol. 97   page: 87   2019

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  13. Reversible bacterial immobilization based on the salt-dependent adhesion of the bacterionanofiber protein AtaA Reviewed

    Yoshimoto Shogo, Ohara Yuki, Nakatani Hajime, Hori Katsutoshi

    MICROBIAL CELL FACTORIES   Vol. 16 ( 1 ) page: 123   2017.7

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Microbial Cell Factories  

    Background: Immobilization of microbial cells is an important strategy for the efficient use of whole-cell catalysts because it simplifies product separation, enables the cell concentration to be increased, stabilizes enzymatic activity, and permits repeated or continuous biocatalyst use. However, conventional immobilization methods have practical limitations, such as limited mass transfer in the inner part of a gel, gel fragility, cell leakage from the support matrix, and adverse effects on cell viability and catalytic activity. We previously showed a new method for bacterial cell immobilization using AtaA, a member of the trimeric autotransporter adhesin family found in Acinetobacter sp. Tol 5. This approach is expected to solve the drawbacks of conventional immobilization methods. However, similar to all other immobilization methods, the use of support materials increases the cost of bioprocesses and subsequent waste materials. Results: We found that the stickiness of the AtaA molecule isolated from Tol 5 cells is drastically diminished at ionic strengths lower than 10 mM and that it cannot adhere in deionized water, which also inhibits cell adhesion mediated by AtaA. Cells immobilized on well plates and polyurethane foam in a salt solution were detached in deionized water by rinsing and shaking, respectively. The detached cells regained their adhesiveness in a salt solution and could rapidly be re-immobilized. The cells expressing the ataA gene maintained their adhesiveness throughout four repeated immobilization and detachment cycles and could be repeatedly immobilized to polyurethane foam by a 10-min shake in a flask. We also demonstrated that both bacterial cells and a support used in a reaction could be reused for a different type of reaction after detachment of the initially immobilized cells from the support and a subsequent immobilization step. Conclusions: We invented a unique reversible immobilization method based on the salt-dependent adhesion of the AtaA molecule that allows us to reuse bacterial cells and supports by a simple manipulation involving a deionized water wash. This mitigates problems caused by the use of support materials and greatly helps to enhance the efficiency and productivity of microbial production processes.

    DOI: 10.1186/s12934-017-0740-7

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  14. Secretion of the Intimin Passenger Domain Is Driven by Protein Folding Reviewed International coauthorship

    Leo Jack C., Oberhettinger Philipp, Yoshimoto Shogo, Udatha D. B. R. K. Gupta, Morth J. Preben, Schuetz Monika, Hori Katsutoshi, Linke Dirk

    JOURNAL OF BIOLOGICAL CHEMISTRY   Vol. 291 ( 38 ) page: 20096 - 20112   2016.9

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    Intimin is an essential adhesin of attaching and effacing organisms such as entropathogenic Escherichia coli. It is also the prototype of type Ve secretion or inverse autotransport, where the extracellular C-terminal region or passenger is exported with the help of an N-terminal transmembrane β-barrel domain. We recently reported a stalled secretion intermediate of intimin, where the passenger is located in the periplasm but the β-barrel is already inserted into the membrane. Stalling of this mutant is due to the insertion of an epitope tag at the very N terminus of the passenger. Here, we examined how this insertion disrupts autotransport and found that it causes misfolding of the N-terminal immunoglobulin (Ig)-like domain D00. We could also stall the secretion by making an internal deletion in D00, and introducing the epitope tag into the second Ig-like domain, D0, also resulted in reduced passenger secretion. In contrast to many classical autotransporters, where a proximal folding core in the passenger is required for secretion, the D00 domain is dispensable, as the passenger of an intimin mutant lacking D00 entirely is efficiently exported. Furthermore, the D00 domain is slightly less stable than the D0 and D1 domains, unfolding at ∼200 piconewtons (pN) compared with ∼250 pN for D0 and D1 domains as measured by atomic force microscopy. Our results support a model where the secretion of the passenger is driven by sequential folding of the extracellular Ig-like domains, leading to vectorial transport of the passenger domain across the outer membrane in an N to C direction.

    DOI: 10.1074/jbc.M116.731497

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  15. Discovery of a novel periplasmic protein that forms a complex with a trimeric autotransporter adhesin and peptidoglycan Reviewed

    Ishikawa Masahito, Yoshimoto Shogo, Hayashi Ayumi, Kanie Junichi, Hori Katsutoshi

    MOLECULAR MICROBIOLOGY   Vol. 101 ( 3 ) page: 394 - 410   2016.8

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Molecular Microbiology  

    Trimeric autotransporter adhesins (TAAs), fibrous proteins on the cell surface of Gram-negative bacteria, have attracted attention as virulence factors. However, little is known about the mechanism of their biogenesis. AtaA, a TAA of Acinetobacter sp. Tol 5, confers nonspecific, high adhesiveness to bacterial cells. We identified a new gene, tpgA, which forms a single operon with ataA and encodes a protein comprising two conserved protein domains identified by Pfam: an N-terminal SmpA/OmlA domain and a C-terminal OmpA_C-like domain with a peptidoglycan (PGN)-binding motif. Cell fractionation and a pull-down assay showed that TpgA forms a complex with AtaA, anchoring it to the outer membrane (OM). Isolation of total PGN-associated proteins showed TpgA binding to PGN. Disruption of tpgA significantly decreased the adhesiveness of Tol 5 because of a decrease in surface-displayed AtaA, suggesting TpgA involvement in AtaA secretion. This is reminiscent of SadB, which functions as a specific chaperone for SadA, a TAA in Salmonella species; however, SadB anchors to the inner membrane, whereas TpgA anchors to the OM through AtaA. The genetic organization encoding the TAA–TpgA-like protein cassette can be found in diverse Gram-negative bacteria, suggesting a common contribution of TpgA homologues to TAA biogenesis.

    DOI: 10.1111/mmi.13398

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  16. An Acinetobacter trimeric autotransporter adhesin reaped from cells exhibits its nonspecific stickiness via a highly stable 3D structure Reviewed

    Yoshimoto Shogo, Nakatani Hajime, Iwasaki Keita, Hori Katsutoshi

    SCIENTIFIC REPORTS   Vol. 6   page: 28020   2016.6

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Scientific Reports  

    Trimeric autotransporter adhesins (TAAs), cell surface proteins of Gram-negative bacteria, mediate bacterial adhesion to host cells and extracellular matrix proteins. However, AtaA, a TAA in the nonpathogenic Acinetobacter sp. strain Tol 5, shows nonspecific, high adhesiveness to abiotic material surfaces as well as to biotic surfaces. AtaA is a homotrimer of polypeptides comprising 3,630 amino acids and forms long nanofibers; therefore, it is too large and structurally complex to be produced as a recombinant protein. In this study, we isolated AtaA's passenger domain (AtaA PSD), which is translocated to the cell surface through the C-terminal transmembrane domain and exhibits biological functions, using a new method. We introduced a protease recognition site and reaped AtaA nanofibers 225 nm in length from the cell surface through proteolytic cleavage with a specific protease. Biochemical and biophysical analyses of the purified native AtaA PSD revealed that it has a stable structure under alkaline and acidic conditions. Temperatures above 80 °C, which disrupted AtaA's higher-order structure but maintained the full-length AtaA polypeptide, inactivated AtaA's nonspecific adhesiveness, suggesting that the stickiness of AtaA requires its 3D structure. This finding refutes the widespread but vague speculation that large unfolded polypeptides readily stick to various surfaces.

    DOI: 10.1038/srep28020

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MISC 2

  1. 小型化接着タンパク質による大腸菌の固定化と微生物反応への応用

    吉本 将悟 , 堀 克敏

    バイオサイエンスとインダストリー   Vol. 82 ( 1 ) page: 20 - 24   2024.1

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  2. 微生物の非特異的な細胞接着

    吉本将悟

    生物工学会誌   Vol. 97 ( 2 ) page: 87 - 87   2019.2

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    Authorship:Lead author, Corresponding author  

Presentations 62

  1. ; 三量体接着タンパク質AtaAのC末端ドメインの力学的安定性の分子メカニズム解析

    笹原 純、加藤 周、吉本将悟、藤本和士、堀 克敏

    2023年度 日本生物工学会中部支部例会  2024.1.23  日本生物工学会中部支部

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    Event date: 2024.1

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  2. Acinetobacter sp. Tol 5の外膜タンパク質AtaAにおけるリシン残基のメチル化修飾

    井上 翔理, 岡 大椰, 吉本 将悟, 堀 克敏

    化学工学会 第54回秋季大会  2023.9.11 

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    Event date: 2023.9

    Presentation type:Poster presentation  

  3. 細胞表層タンパク質AtaAの膜貫通ドメインを用いたナノポアの構築

    黄 泰賢, 吉本 将悟, 笹原 純, 川野 竜司, 堀 克敏

    化学工学会 第54回秋季大会  2023.9.11 

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    Event date: 2023.9

    Presentation type:Poster presentation  

  4. 人工タンパク質繊維に向けた繊維状タンパク質重合反応の基礎検討

    長谷 彩沙, 南畑 孝介, 石川 聖人, 吉本 将悟, 堀 克敏, 神谷 典穂

    化学工学会 第54回秋季大会  2023.9.11 

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    Event date: 2023.9

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  5. 分子動力学シミュレーションによる三量体接着タンパク質AtaAの力学的安定性の分子機構解析

    笹原 純, 吉本 将悟, 藤本 和士, 堀 克敏

    化学工学会 第54回秋季大会  2023.9.11 

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    Event date: 2023.9

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  6. 原子間力顕微鏡と分子動力学計算による細菌ファイバータンパク質AtaA の力学強度解析

    笹原 純, 加藤 周, 吉本 将悟, 藤本 和士, 堀 克敏

    第17回バイオ関連化学シンポジウム  2023.9.9 

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    Event date: 2023.9

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  7. 接着タンパク質AtaAの機能部位特定と小型AtaAを用いた大腸菌の迅速簡便な固定化

    吉本 将悟, 石川 聖人, 鈴木 淳巨, Linke Dirk, Lupas Andrei, 堀 克敏

    第17回バイオ関連化学シンポジウム  2023.9.9 

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    Event date: 2023.9

    Presentation type:Oral presentation (general)  

  8. Identification of the adhesive domain of AtaA from Acinetobacter sp. Tol 5 and its application in immobilizing Escherichia coli International conference

    Shogo Yoshimoto

    CIS-BIO 2023  2023.9.6 

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    Event date: 2023.9

    Presentation type:Poster presentation  

  9. Experimental and computational approaches to the molecular characterization of a bacterial adhesin protein Invited

    Katsutoshi Hori, Jun Sasahara, Amane Kato, Shogo Yoshimoto

    2023.9.5 

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    Event date: 2023.9

    Presentation type:Oral presentation (invited, special)  

  10. アミノ酸物性値を用いたSARS-CoV-2スパイク配列の高速特徴抽出法の開発

    岡 大椰, 吉本 将悟, 堀 克敏

    第75回日本生物工学会大会  2023.9.3  日本生物工学会

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    Event date: 2023.9

    Presentation type:Oral presentation (general)  

  11. 原子間力顕微鏡を用いた三量体接着タンパク質AtaAの1分子強靭性発現メカニズム解明

    加藤 周, 笹原 純, 吉本 将悟, 堀 克敏

    第75回日本生物工学会大会  2023.9.3  日本生物工学会

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    Event date: 2023.9

    Presentation type:Oral presentation (general)  

  12. 繊維状タンパク質の両末端連結反応を基盤とした人工繊維の形成

    長谷 彩沙, 南畑 孝介, 石川 聖人, 吉本 将悟, 堀 克敏, 神谷 典穂

    第75回日本生物工学会大会  2023.9.3  日本生物工学会

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    Event date: 2023.9

    Presentation type:Oral presentation (general)  

  13. トルエン資化細菌Acinetobacter sp. Tol 5の外膜タンパク質におけるメチル化修飾

    井上 翔理, 岡 大椰, 吉本 将悟, 堀 克敏

    第75回日本生物工学会大会  2023.9.3  日本生物工学会

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    Event date: 2023.9

    Presentation type:Oral presentation (general)  

  14. トルエン資化細菌Acinetobacter sp. Tol 5の外膜トランスポーター機能解析

    森 さくら, 吉本 将悟, 岡 大椰, 堀 克敏

    第75回日本生物工学会大会  2023.9.3  日本生物工学会

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    Event date: 2023.9

    Presentation type:Oral presentation (general)  

  15. 全原子分子動力学計算による三量体接着タンパク質AtaAの機械的安定性メカニズムの解析

    笹原 純, 吉本 将悟, 藤本 和士, 堀 克敏

    第75回日本生物工学会大会  2023.9.4  日本生物工学会

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    Event date: 2023.9

    Presentation type:Oral presentation (general)  

  16. 三量体型オートトランスポーターアドへシンの膜貫通ドメインを用いたナノポアの構築

    黄 泰賢, 吉本 将悟, 笹原 純, 川野 竜司, 堀 克敏

    第75回日本生物工学会大会  2023.9.4  日本生物工学会

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    Event date: 2023.9

    Presentation type:Oral presentation (general)  

  17. 膜内外に異種タンパク質を連結したバクテリアサイズユニラメラリポソームの簡便な作製法

    野場 考策, 吉本 将悟, 田中 良和, 横山 武志, 松浦 友亮, 堀 克敏

    第75回日本生物工学会大会  2023.9.5  日本生物工学会

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    Event date: 2023.9

    Presentation type:Oral presentation (general)  

  18. 三量体接着タンパク質AtaAの小型化とそれを用いた大腸菌の固定化

    吉本 将悟, 青木 壮太, 小原 優季, 石川 聖人, 鈴木 淳巨, Linke Dirk, Lupas Andrei, 堀 克敏

    第75回日本生物工学会大会  2023.9.5  日本生物工学会

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    Event date: 2023.9

    Presentation type:Oral presentation (general)  

  19. Immobilization of biocatalysts on solid surfaces using the bacterionanofiber protein AtaA International conference

    S. Yoshimoto, M. Sawada, K. Noba, M. Ishikawa, K. Hori

    The 10th Congress of European Microbiologists (FEMS2023)   2023.7.11 

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    Event date: 2023.7

    Language:English   Presentation type:Poster presentation  

    Venue:Hamburg   Country:Germany  

  20. Mechanical stability analysis of an Acinetobacter adhesin by steered molecular dynamics simulation International coauthorship International conference

    J. Sasahara, K. Fujimoto, S. Yoshimoto, D. Linke, A.N. Lupas, K. Hori

    The 10th Congress of European Microbiologists (FEMS2023)   2023.7.11 

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    Event date: 2023.7

    Language:English   Presentation type:Poster presentation  

    Venue:Hamburg   Country:Germany  

  21. Single cell adhesion force mapping of a highly sticky bacterium in liquid International conference

    K.Hori, S. Yoshimoto

    The 10th Congress of European Microbiologists (FEMS2023)   2023.7.12 

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    Event date: 2023.7

    Language:English   Presentation type:Poster presentation  

    Venue:Hamburg   Country:Germany  

  22. 細胞表層タンパク質AtaA_Cheadのナノ力学応答解析

    笹原 純, 加藤 周, 橋本貴幸, 吉本将悟, 藤本和士, 堀 克敏

    生物工学若手研究者の集い 夏のセミナー2023  2023.6.24 

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    Event date: 2023.6

    Presentation type:Poster presentation  

  23. Acinetobacter sp. Tol 5の外膜タンパク質AtaAにおけるメチル化修飾の解析

    井上翔理, 岡 大椰, 吉本将悟, 堀 克敏

    生物工学若手研究者の集い 夏のセミナー2023  2023.6.24 

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    Event date: 2023.6

    Presentation type:Poster presentation  

  24. 組換え体膜タンパク質を用いたナノポアの創出と特性解析

    黄 泰賢, 吉本将悟, 笹原 純, 川野竜司, 堀 克敏

    生物工学若手研究者の集い 夏のセミナー2023  2023.6.24 

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    Event date: 2023.6

    Presentation type:Poster presentation  

  25. Single-cell analysis of adhesive bacteria using atomic force microscopy Invited International conference

    Shogo Yoshimoto

    MISC symposium in Oslo 2023  2023.2.6 

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    Event date: 2023.2

    Language:English   Presentation type:Oral presentation (invited, special)  

    Venue:Oslo   Country:Norway  

  26. Mechanical stability analysis of a cell surface protein AtaA_Chead by steered molecular dynamics simulation and atomic force microscopy International conference

    J. Sasahara, K. Fujimoto, T. Hashimoto, A. Kato, S. Yoshimoto, K. Hori

    MISC symposium in Oslo 2023   2023.2 

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    Event date: 2023.2

    Presentation type:Poster presentation  

    Venue:Oslo   Country:Norway  

  27. Bottom-up creation of an artificial cell covered with the adhesive bacterionanofiber protein AtaA International conference

    Kosaku Noba ,Masahito Ishikawa ,Atsuko Uyeda ,Takayoshi Watanabe ,Takahiro Hohsaka ,Shogo Yoshimoto ,Tomoaki Matsuura ,Katsutoshi Hori

    MISC symposium in Oslo 2023   2023.2 

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    Event date: 2023.2

    Presentation type:Poster presentation  

    Venue:Oslo   Country:Norway  

  28. MD計算を用いた細胞表層タンパク質AtaA_Cheadの力学応答解析

    笹原 純、橋本貴幸、藤本和士、吉本将悟、堀 克敏

    2022年度 日本生物工学会中部支部例会  2022.9.22 

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    Event date: 2022.9

    Presentation type:Oral presentation (general)  

  29. 原子間力顕微鏡を用いた高付着性細菌の1細胞・1分子付着力解析

    第16回バイオ関連化学シンポジウム  2022.9 

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    Event date: 2022.9

    Presentation type:Oral presentation (general)  

  30. 人工細菌を志向した非対称なタンパク質組成を有する細菌サイズのリポソーム創出

    第16回バイオ関連化学シンポジウム  2022.9 

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    Event date: 2022.9

    Presentation type:Poster presentation  

  31. 分子動力学シミュレーションによる細胞表層タンパク質AtaA_Cheadの力学応答解析

    第16回バイオ関連化学シンポジウム  2022.9 

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    Event date: 2022.9

    Presentation type:Poster presentation  

  32. プラスチックの分解速度向上を目指した酵素複合体の構築

    第16回バイオ関連化学シンポジウム  2022.9 

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    Event date: 2022.9

    Presentation type:Poster presentation  

  33. 原子間力顕微鏡を用いたAtaAのC末端パッセンジャードメインの1分子強靭性解析

    第16回バイオ関連化学シンポジウム  2022.9 

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    Event date: 2022.9

    Presentation type:Poster presentation  

  34. 高付着性細菌の定量的付着力解析

    第15回バイオ関連化学シンポジウム  2021.9 

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    Event date: 2021.9

    Presentation type:Poster presentation  

  35. タンパク質の配向を制御した再構成型リポソームの作製

    第15回バイオ関連化学シンポジウム  2021.9 

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    Event date: 2021.9

    Presentation type:Poster presentation  

  36. AFMを用いたファイバータンパク質AtaAの力学特性解析法の確立

    第15回バイオ関連化学シンポジウム  2021.9 

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    Event date: 2021.9

    Presentation type:Poster presentation  

  37. PET加水分解酵素と高付着性タンパク質の連結体構築

    第15回バイオ関連化学シンポジウム  2021.9 

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    Event date: 2021.9

    Presentation type:Poster presentation  

  38. The trimeric autotransporter adhesin of the highly adhesive Acinetobacter strain Tol 5

    第94回日本細菌学会総会  2021 

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    Event date: 2021

    Presentation type:Oral presentation (invited, special)  

  39. 正確なドメイン欠損による巨大ファイバータンパク質の再構成

    化学工学会第85年会  2020 

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    Event date: 2020

    Presentation type:Poster presentation  

  40. 発現ホストの違いによる高凝集性タンパク質AtaAの付着凝集特性の相違

    化学工学会第85年会  2020 

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    Event date: 2020

    Presentation type:Poster presentation  

  41. 高付着性細菌Acinetobacter sp. Tol 5の難付着性材料に対する付着性の評価

    化学工学会第84年会  2019 

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    Event date: 2019

  42. バクテリオナノファイバー蛋白質AtaAで修飾した接着性人工細胞の創出

    日本化学会秋季事業 第9回CSJ化学フェスタ2019  2019 

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    Event date: 2019

    Presentation type:Poster presentation  

  43. AFMを用いた高付着性細菌及びその細胞表層タンパク質の接着力測定

    第71回日本生物工学会大会  2019 

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    Event date: 2019

    Presentation type:Oral presentation (general)  

  44. バクテリオナノファイバー蛋白質AtaAで修飾した接着性人工細胞の創出

    第13回バイオ関連化学シンポジウム  2019 

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    Event date: 2019

    Presentation type:Poster presentation  

  45. 高付着性蛋白質AtaAで被覆された人工細胞のボトムアップ創成

    2019年度 生物工学会中部支部例会  2019 

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    Event date: 2019

    Presentation type:Oral presentation (general)  

  46. 高付着性微生物及び表層タンパク質の原子間力顕微鏡解析

    化学工学会第84年会  2019 

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    Event date: 2019

  47. 高接着性蛋白質AtaAの反復配列を利用した部分欠損体の構築と機能ドメイン探索

    2018 

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    Event date: 2018

  48. ナノファイバータンパク質AtaAの接着阻害物質の探索と微生物固定化制御

    化学工学会 第50回秋季大会  2018 

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    Event date: 2018

  49. ナノファイバータンパク質AtaAによる全細胞触媒の可逆的固定化

    化学工学会 第50回秋季大会  2018 

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    Event date: 2018

  50. Acinetobacter sp. Tol 5の難付着性材料に対する付着性の評価

    第70回日本生物工学会大会  2018 

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    Event date: 2018

  51. ナノファイバータンパク質AtaAの塩濃度依存的な接着性を利用した細菌の可逆的固定化

    第70回日本生物工学会大会  2018 

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    Event date: 2018

  52. Structural insight into the highly adhesive protein AtaA

    IGER International Symposium on Cell Surface Structures and Functions  2017 

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    Event date: 2017

  53. Molecular insight into a periplasmic protein interacting with trimeric autotransporter adhesin and peptidoglycan

    Protein Secretion in Bacteria Conference  2016 

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    Event date: 2016

  54. 接着性ナノファイバータンパク質AtaAを介した微生物付着の阻害物質についての検討

    日本生物工学会大会  2015 

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    Event date: 2015

  55. Biophysical characterization of a trimeric autotransporter adhesin, AtaA, from the highly adhesive bacterium Acinetobacter sp. Tol 5

    The 6th Congress of European Microbiologists  2015 

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    Event date: 2015

  56. Characterization of AtaA, an Acinetobacter TAA, immobilizing bacterial cells firmly to various surfaces

    4th International Symposium of the SFB766  2015 

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    Event date: 2015

  57. Molecular characterization of a trimeric autotransporter adhesin from Acinetobacter sp. Tol 5 mediating high adhesiveness to various abiotic surfaces

    4th International SFB 766 Symposium  2015 

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    Event date: 2015

  58. 接着性ナノファイバータンパク質AtaAと共発現するタンパク質との相互作用

    日本生物工学会  2015 

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    Event date: 2015

  59. Characterization of a trimeric autotransporter adhesin from a highly adhesive bacterium Acinetobacter sp. Tol 5

    III International Conference on Antimicrobial Research  2014 

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    Event date: 2014

  60. AtaA, a highly adhesive bacterionanofiber, mediates ionic strength dependent-bacterial adhesion

    The 5th Congress of European Microbiologists  2013 

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    Event date: 2013

  61. Reversible bacterial immobilization based on the adhesive property of a novel adhesin from Acinetobacter sp. Tol 5

    IGER International Symposium on Cell Surface Structures and Functions  2013 

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    Event date: 2013

  62. Adhesion profiles of Acinetobacter sp. Tol 5 mediated by nanofiber

    2012 International Symposium on Advanced Biological Engineering  2012 

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    Event date: 2012

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Research Project for Joint Research, Competitive Funding, etc. 1

  1. 固相基質分解酵素複合体の分子設計基盤の確立

    Grant number:JPMJAX20BL  2020.12 - 2023.3

    JST  ACT-X 

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    Authorship:Principal investigator 

KAKENHI (Grants-in-Aid for Scientific Research) 3

  1. 水溶性遊離分子を基質とする酵素による高分子材料の分解

    Grant number:22K04839  2022.4 - 2025.3

    科学研究費助成事業  基盤研究(C)

    吉本 将悟

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    Authorship:Principal investigator 

    Grant amount:\4030000 ( Direct Cost: \3100000 、 Indirect Cost:\930000 )

    プラスチックやゴムといった高分子材料の処理方法として酵素による分解が注目されている。しかし酵素のうち、高分子材料を分解できるものはごくわずかである。本研究では、元来水溶性遊離分子を基質とする酵素をタンパク質工学的に改変することで、高分子材料を分解できるようにすることを目指す。

  2. Analysis of the mechanical property of nanofiber protein AtaA

    Grant number:20K15098  2020.4 - 2022.3

    Grants-in-Aid for Scientific Research  Grant-in-Aid for Early-Career Scientists

    Yoshimoto Shogo

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    Authorship:Principal investigator 

    Grant amount:\4290000 ( Direct Cost: \3300000 、 Indirect Cost:\990000 )

    In this study, we analyzed the nanomechanical properties of the nanofiber protein AtaA using atomic force microscopy and molecular dynamics simulations with the aim of elucidating the mechanism of viscoelasticity of AtaA. As a result, we found that tough domains exist in AtaA, and that these mechanical properties are based on the precisely designed three-dimensional structure of the protein.

  3. Elucidation of the adhesion mechanism of the non-specific adhesion protein AtaA

    Grant number:18K14062  2018.4 - 2021.3

    Grants-in-Aid for Scientific Research  Grant-in-Aid for Early-Career Scientists

    Yoshimoto Shogo

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    Authorship:Principal investigator 

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

    The purpose of this study is clarification of the adhesion mechanism of AtaA, which exhibits non-specific and high adhesiveness. Through the analysis of the functional sites and the adhesion-driving force of AtaA, the following results were obtained. (1) Adhesion analysis using a quartz crystal microbalance (QCM) revealed the adhesion-driving force of AtaA. (2) By constructing mutants of AtaA and evaluating their adhesive properties, the functional sites were determined. (3) Based on the above findings, we succeeded in reconstructing a protein that exhibits AtaA-like adhesion properties.

 

Teaching Experience (On-campus) 7

  1. 生命システム工学セミナー1A, 1C, 2E

    2023

  2. 生命システム工学特別実験及び演習1,3

    2023

  3. 卒業研究A

    2023

  4. 化学生命工学演習1

    2023

  5. 生命システム工学特別実験及び演習2,4

    2022

  6. 生命システム工学セミナー 1B,1D,2D

    2022

  7. 卒業研究B

    2022

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Academic Activities 3

  1. CIS-BIO 2023 事務局 International contribution

    Role(s):Planning, management, etc.

    日本生物工学会中部支部  2023.9

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  2. 第75回日本生物工学会大会 国際シンポジウムオーガナイザー International contribution

    Role(s):Panel moderator, session chair, etc.

    2023.9

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  3. 第75回日本生物工学会大会 大会実行委員

    Role(s):Planning, management, etc.

    2023.9

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    Type:Academic society, research group, etc.