研究者詳細 - 堀　克敏

論文 - 堀　克敏

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Bottom-up Creation of an Artificial Cell Covered with the Adhesive Bacterionanofiber Protein AtaA 査読有り国際誌 Open Access

Noba, K; Ishikawa, M; Uyeda, A; Watanabe, T; Hohsaka, T; Yoshimoto, S; Matsuura, T; Hori, K

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 141 巻 ( 48 ) 頁： 19058 - 19066 2019年12月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：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

Web of Science

Scopus

PubMed

細菌の接着機構と界面微生物工学プロセスの創出に関する研究招待有り

堀　克敏

生物工学会誌第102巻巻 ( 第2号 ) 頁： 54 - 60 2024年

微生物飼料を活用した次世代陸上養殖の展開

田丸　浩、堀　克敏

月刊バイオインダストリー 40 巻 ( 1 ) 頁： 51 - 57 2023年1月

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; バイオサイエンスとインダストリー

吉本　将悟、堀　克敏

バイオサイエンスとインダストリー 40 巻頁： 51 - 57 2023年

Perturbation by Antimicrobial Bacteria of the Epidermal Bacterial Flora of Rainbow Trout in Flow-Through Aquaculture 査読有り国際誌 Open Access

Nakatani, H; Yamada, N; Hashimoto, N; Okazaki, F; Arakawa, T; Tamaru, Y; Hori, K

BIOLOGY-BASEL 11 巻 ( 8 ) 頁： 1249 2022年8月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Biology

The bacterial flora of the epidermal mucus of fish is closely associated with the host’s health and susceptibility to pathogenic infections. In this study, we analyzed the epidermal mucus bacteria of rainbow trout (Oncorhynchus mykiss) reared in flow-through aquaculture under environmental perturbations. Over ~2 years, the bacteria present in the skin mucus and water were analyzed based on the 16S rDNA sequences. The composition of the mucus bacterial community showed significant monthly fluctuations, with frequent changes in the dominant bacterial species. Analysis of the beta- and alpha-diversity of the mucus bacterial flora showed the fluctuations of the composition of the flora were caused by the genera Pseudomonas, Yersinia, and Flavobacterium, and some species of Pseudomonas and Yersinia in the mucus were identified as antimicrobial bacteria. Examination of the antimicrobial bacteria in the lab aquarium showed that the natural presence of antimicrobial bacteria in the mucus and water, or the purposeful addition of them to the rearing water, caused a transition in the mucus bacteria community composition. These results demonstrate that specific antimicrobial bacteria in the water or in epidermal mucus comprise one of the causes of changes in fish epidermal mucus microflora.

DOI： 10.3390/biology11081249

The extracellular juncture domains of Type 5 autotransporters 査読有り国際共著

Weikum, J; Kulakova, A; Tesei, G; Yoshimoto, S; Jægerum, LV; Schütz, M; Hori, K; Skepö, M; Harris, P; Leo, JC; Morth, JP

ACTA CRYSTALLOGRAPHICA A-FOUNDATION AND ADVANCES 78 巻頁： E5 - E5 2022年8月

Fabrication of a highly protective 3D-printed mask and evaluation of its viral filtration efficiency using a human head mannequin 査読有り国際誌 Open Access

Ohara, Y; Kanie, J; Hori, K

HARDWAREX 11 巻頁： e00314 2022年4月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：HardwareX

Facemasks are one of the most effective and low-cost prophylactics for COVID-19. In the spring 2020, when a severe shortage of facemasks occurred worldwide, various types of 3D-printed masks were designed and proposed. However, the protective effects conferred by most of these masks were not experimentally evaluated. Here, we provide a new simple design of 3D-printed mask and evaluate its protective effect in a viral filtration test using a human head mannequin. The developed mask can be constructed with a low-cost 3D printer, with an approximate production cost of US $4. This mask has three parts: the main part, wearing parts, and a piece of non-woven fabric filter. The volume of the filter, which needs to be changed daily, was reduced to approximately 1/10 of that of commercially available surgical masks used in this study. The developed mask is fabricated from polylactic acid, a biodegradable plastic, and its surface contour contacting the face may be adjusted after softening the material with hot water at 60–80 °C. The viral filtration efficiency of the developed mask was found to be over 80%. This performance is better than that of commercially available facemasks, such as surgical masks and cloth masks, and equal to those of KN95 and KF94.

DOI： 10.1016/j.ohx.2022.e00314

細菌接着タンパク質と微生物固定化への利用招待有り

堀　克敏

バイオインダストリー 39 巻 ( 3 ) 頁： 11 - 18 2022年3月

Recent advances in research on biointerfaces: From cell surfaces to artificial interfaces 招待有り査読有り国際誌

K. Hori, S. Yoshimoto, T. Yoshino, T. Zako, G. Hirao, S. Fujita, C. Nakamura, A. Yamagishi, and N. Kamiya

Journal of Bioscience and Bioengineering 133 巻 ( 3 ) 頁： 195 - 207 2022年1月

担当区分：筆頭著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

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 lipideprotein 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.

ウイルス除去効果の高い樹脂製立体マスクの開発招待有り

堀　克敏

繊維機械学会誌月刊せんい 74 巻 ( 11 ) 頁： 593 - 598 2021年11月

Complete Genome Sequence of the Highly Adhesive Bacterium Acinetobacter sp. Strain Tol 5 査読有り国際誌 Open Access

Ishikawa, M; Hori, K

MICROBIOLOGY RESOURCE ANNOUNCEMENTS 10 巻 ( 35 ) 頁： e0056721 2021年9月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Microbiology Resource Announcements

Acinetobacter sp. strain Tol 5 is a nonpathogenic Gram-negative bacterium with biotechnological and environmental applications. Here, we report the complete genome sequence of Acinetobacter sp. strain Tol 5, which has a genome size of 4,799,506 bp and a G1C content of 38.1%.

DOI： 10.1128/MRA.00567-21

界面微生物工学微生物固定化・細胞表層工学から膜タンパク質利用技術まで招待有り

堀　克敏

化学と生物 59 巻 ( 8 ) 頁： 393 - 400 2021年8月

Switching Between Methanol Accumulation and Cell Growth by Expression Control of Methanol Dehydrogenase in Methylosinus trichosporium OB3b Mutant 査読有り国際誌 Open Access

Ito, H; Yoshimori, K; Ishikawa, M; Hori, K; Kamachi, T

FRONTIERS IN MICROBIOLOGY 12 巻頁： 639266 2021年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Frontiers in Microbiology

Methanotrophs have been used to convert methane to methanol at ambient temperature and pressure. In order to accumulate methanol using methanotrophs, methanol dehydrogenase (MDH) must be downregulated as it consumes methanol. Here, we describe a methanol production system wherein MDH expression is controlled by using methanotroph mutants. We used the MxaF knockout mutant of Methylosinus trichosporium OB3b. It could only grow with MDH (XoxF) which has a cerium ion in its active site and is only expressed by bacteria in media containing cerium ions. In the presence of 0 μM copper ion and 25 μM cerium ion, the mutant grew normally. Under conditions conducive to methanol production (10 μM copper ion and 0 μM cerium ion), cell growth was inhibited and methanol accumulated (2.6 μmol·mg−1 dry cell weight·h−1). The conversion efficiency of the accumulated methanol to the total amount of methane added to the reaction system was ~0.3%. The aforementioned conditions were repeatedly alternated by modulating the metal ion composition of the bacterial growth medium.

DOI： 10.3389/fmicb.2021.639266

微生物接着タンパク質AtaAの無限の可能性招待有り

堀　克敏

バイオインダストリー 38 巻 ( 2 ) 頁： 8 - 17 2021年2月

Establishing a Percutaneous Infection Model Using Zebrafish and a Salmon Pathogen 査読有り Open Access

Nakatani, H; Hori, K

BIOLOGY-BASEL 10 巻 ( 2 ) 頁： 1 - 15 2021年2月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Biology

To uncover the relationship between skin bacterial flora and pathogen infection, we developed a percutaneous infection model using zebrafish and Yersinia ruckeri, a pathogen causing enteric redmouth disease in salmon and in trout. Pathogen challenge, either alone or together with pricking by a small needle, did not cause infection of the fish. However, cold stress given by water temperature shift from the optimum 28°C for zebrafish to 20°C caused fatal infection of injured fish following pathogen challenge. We investigated the effects of cold stress, injury, and pathogen challenge, alone and in combination, on fish skin bacterial flora using 16S rDNA metagenomics. We found that cold stress drastically altered the skin bacterial flora, which was dominated by Y. ruckeri on infected fish. In addition, fish whose intrinsic skin bacterial flora was disrupted by antibiotics had their skin occupied by Y. ruckeri following a challenge with this pathogen, although the fish survived without injury to create a route for invasion into the fish body. Our results suggest that the intrinsic skin bacterial flora of fish protects them from pathogen colonization, and that its disruption by stress allows pathogens to colonize and dominate their skin.

DOI： 10.3390/biology10020166

The extracellular juncture domains in the intimin passenger adopt a constitutively extended conformation inducing restraints to its sphere of action 査読有り国際共著国際誌 Open Access

Weikum, J; Kulakova, A; Tesei, G; Yoshimoto, S; Jægerum, LV; Schütz, M; Hori, K; Skepö, M; Harris, P; Leo, JC; Morth, JP

SCIENTIFIC REPORTS 10 巻 ( 1 ) 頁： 21249 2020年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：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

人工細胞と固体表面をつなぐ接着ナノファイバー～新規生体触媒の開発を目指して～招待有り

石川　聖人,堀　克敏

バイオサイエンスとインダストリー 78 巻 ( 5 ) 頁： 408 - 409 2020年9月

Extracellular electron transfer mediated by a cytocompatible redox polymer to study the crosstalk among the mammalian circadian clock, cellular metabolism, and cellular redox state 査読有り国際誌 Open Access

Ishikawa, M; Kawai, K; Kaneko, M; Tanaka, K; Nakanishi, S; Hori, K

RSC ADVANCES 10 巻 ( 3 ) 頁： 1648 - 1657 2020年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：RSC Advances

The circadian clock is an endogenous biological timekeeping system that controls various physiological and cellular processes with a 24 h rhythm. The crosstalk among the circadian clock, cellular metabolism, and cellular redox state has attracted much attention. To elucidate this crosstalk, chemical compounds have been used to perturb cellular metabolism and the redox state. However, an electron mediator that facilitates extracellular electron transfer (EET) has not been used to study the mammalian circadian clock due to potential cytotoxic effects of the mediator. Here, we report evidence that a cytocompatible redox polymer pMFc (2-methacryloyloxyethyl phosphorylcholine-co-vinyl ferrocene) can be used as the mediator to study the mammalian circadian clock. EET mediated by oxidized pMFc (ox-pMFc) extracted intracellular electrons from human U2OS cells, resulting in a longer circadian period. Analyses of the metabolome and intracellular redox species imply that ox-pMFc receives an electron from glutathione, thereby inducing pentose phosphate pathway activation. These results suggest novel crosstalk among the circadian clock, metabolism, and redox state. We anticipate that EET mediated by a redox cytocompatible polymer will provide new insights into the mammalian circadian clock system, which may lead to the development of new treatments for circadian clock disorders.

DOI： 10.1039/c9ra10023g

接着タンパク質・接着微生物を利用した効率的バイオプロセスの構築招待有り

石川　聖人，堀　克敏

酵素工学ニュース 84 巻頁： 13 - 17 2020年

接着性人工細胞触媒のボトムアップ創製招待有り

石川　聖人，堀　克敏

生物工学 98 巻頁： 651 - 654 2020年

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