2014年11月, BCSJ賞(日本化学会･欧文誌編集委員会), 日本化学会欧文誌(Bulletin of the Chemical Society of Japan)に掲載の論文,Kinetic analysis of inactivation and enzyme reaction of oxygen-tolerant [NiFe]-hydrogenase at direct electron transfer-type bioanode. Bull. Chem. Soc. Jpn., 87 (11): 1177-1185 (2014), 日本化学会及び欧文誌編集委員会
So;K.;Kitazumi;Y.;Shirai;O.;Kurita;K.;Higuchi;Y.;Kano;K.

Growth and composition analyses of hydrogen oxidizing bacteria Hydrogenovibrio marinus MH-110 in a natural seawater based medium: preliminary study to improve production profitability　　　　　　　　　　　　　　　
Kazuki Ohta; Hiromi Oku; Masaharu Tokuda; Hiroyuki Matsunari; Hirofumi Furuita; Hazuki Yoshinaga; Takeshi Yamamoto; Koji Murashita; Hirofumi Nishihara
Fisheries Sci., 2025年01月, ［査読有り］

水素細菌Hydrogenovibrio marinus MH-110の養魚飼料としての生産および利用に向けた培養、成分分析および短期飼育による特性評価　　　　　　　　　　　　　　　
奥宏海; 西原宏史; 上谷はる; 松成宏之; 徳田雅治; 吉永葉月; 古板博文; 村下幸司; 山本剛史
水産技術, 2024年, ［査読有り］

Enhanced supply of acetyl-CoA by exogenous pantothenate kinase promotes synthesis of poly(3-hydroxybutyrate)
Microbial Cell Factories, 2023年04月20日, ［査読有り］

Structural basis of the redox switches in the NAD-reducing soluble [NiFe]-hydrogenase
Shomura; Y.; Taketa; M.; Nakashima; H.; Tai; H.; Nakagawa; H.; Ikeda; Y.; Ishii; M.; Igarashi; Y.; Nishihara; H.; Yoon; K. S.; Ogo; S.; Hirota; S.; Higuchi; Y., NAD(+) (oxidized form of NAD: nicotinamide adenine dinucleotide)-reducing soluble [ NiFe]hydrogenase (SH) is phylogenetically related to NADH (reduced form of NAD(+)): quinone oxidoreductase (complex I), but the geometrical arrangements of the subunits and Fe-S clusters are unclear. Here, we describe the crystal structures of SH in the oxidized and reduced states. The cluster arrangement is similar to that of complex I, but the subunits orientation is not, which supports the hypothesis that subunits evolved as prebuilt modules. The oxidized active site includes a six-coordinate Ni, which is unprecedented for hydrogenases, whose coordination geometry would prevent O-2 from approaching. In the reduced state showing the normal active site structure without a physiological electron acceptor, the flavin mononucleotide cofactor is dissociated, which may be caused by the oxidation state change of nearby Fe-S clusters and may suppress production of reactive oxygen species., AMER ASSOC ADVANCEMENT SCIENCE
Science, 2017年09月, ［査読有り］

Structural and functional insights into thermally stable cytochrome c' from a thermophile
Sotaro Fujii; Hiroya Oki; Kazuki Kawahara; Daisuke Yamane; Masaru Yamanaka; Takahiro Maruno; Yuji Kobayashi; Misa Masanari; Satoshi Wakai; Hirofumi Nishihara; Tadayasu Ohkubo; Yoshihiro Sambongi, Thermophilic Hydrogenophilus thermoluteolus cytochrome c (PHCP) exhibits higher thermal stability than a mesophilic counterpart, Allochromatium vinosum cytochrome c (AVCP), which has a homo-dimeric structure and ligand-binding ability. To understand the thermal stability mechanism and ligand-binding ability of the thermally stable PHCP protein, the crystal structure of PHCP was first determined. It formed a homo-dimeric structure, the main chain root mean square deviation (rmsd) value between PHCP and AVCP being 0.65 angstrom. In the PHCP structure, six specific residues appeared to strengthen the heme-related and subunit-subunit interactions, which were not conserved in the AVCP structure. PHCP variants having altered subunit-subunit interactions were more severely destabilized than ones having altered heme-related interactions. The PHCP structure further revealed a ligand-binding channel and a penta-coordinated heme, as observed in the AVCP protein. A spectroscopic study clearly showed that some ligands were bound to the PHCP protein. It is concluded that the dimeric PHCP from the thermophile is effectively stabilized through heme-related and subunit-subunit interactions with conservation of the ligand-binding ability.
Brief SummaryWe report the X-ray crystal structure of cytochrome c (PHCP) from thermophilic Hydrogenophilus thermoluteolus. The high thermal stability of PHCP was attributed to heme-related and subunit-subunit interactions, which were confirmed by a mutagenesis study. The ligand-binding ability of PHCP was examined by spectrophotometry. PHCP acquired the thermal stability with conservation of the ligand-binding ability. This study furthers the understanding of the stability and function of cytochromes c., WILEY
PROTEIN SCIENCE, 2017年04月, ［査読有り］

Transcriptome profiles of central carbon metabolism under autotrophic, heterotrophic, and mixotrophic conditions in Hydrogenophilus thermoluteolus TH-1　　　　　　　　　　　　　　　
Nguyen, H. T; Hirano, S; Arai, H; Nishihara, H; Ishii, M
J. Jpn. Soc. Extremophiles, 2017年, ［査読有り］

Bioelectrochemical analysis of thermodynamics of the catalytic cycle and kinetics of the oxidative inactivation of oxygen-tolerant [NiFe]-hydrogenase
Keisei So; Rui Hamamoto; Ryosuke Takeuchi; Yuki Kitazumi; Osamu Shirai; Ryohei Endo; Hirofumi Nishihara; Yoshiki Higuchi; Kenji Kano, Membrane-bound [NiFe]-hydrogenase from Hydrogenovibrio marinus (HmMBH) is an O-2-tolerant enzyme and allows direct electron transfer (DET)-type bioelectrocatalysis for the H-2 oxidation. Very fast interfacial electron transfer occurs between the [NiFe]-active site of HmMBH and the electrode, and the potential dependence of the steady-state DET-type catalytic current has been analyzed on a thermodynamic model of a two-step one electron transfer to get a Pourbaix diagram of the catalytic center. A reversible and oxidative inactivation that occurs when the [NiFe]-hydrogenases are suffering from the oxidative stress at high electrode potentials or high solution potentials has been kinetically analyzed for the time-dependence of the steady-state catalytic current as a measure. The kinetic analysis has shown that the rate-determining step of the oxidative inactivation is not electrochemical but chemical process and that the rate of the reductive reactivation is determined by the electrochemical process. The observed catalytic waves, especially the dependence of the waves on the scan rate and the hydrogen concentration, have been well reproduced by simulation with the thermodynamic and kinetic parameters evaluated here. (C) 2016 Elsevier B.V. All rights reserved., ELSEVIER SCIENCE SA
JOURNAL OF ELECTROANALYTICAL CHEMISTRY, 2016年04月, ［査読有り］

Improved purification, crystallization and crystallographic study of Hyd-2-type [NiFe]-hydrogenase from Citrobacter sp S-77
Noor Dina Muhd Noor; Koji Nishikawa; Hirofumi Nishihara; Ki-Seok Yoon; Seiji Ogo; Yoshiki Higuchi, The purification procedure of Hyd-2-type [NiFe]-hydrogenase from Citrobacter sp. S-77 was improved by applying treatment with trypsin before chromatography. Purified protein samples both with and without trypsin treatment were successfully crystallized using the sitting-drop vapour-diffusion method with polyethylene glycol as a precipitant. Both crystals belonged to space group P2(1), with unit-cell parameters a = 63.90, b = 118.89, c = 96.70 angstrom, beta = 100.61 degrees for the protein subjected to trypsin treatment and a = 65.38, b = 121.45, c = 98.63 angstrom, beta = 102.29 degrees for the sample not treated with trypsin. The crystal obtained from the trypsin-treated protein diffracted to 1.60 angstrom resolution, which is considerably better than the 2.00 angstrom resolution obtained without trypsin treatment. The [NiFe]-hydrogenase from Citrobacter sp. S-77 retained catalytic activity with some amount of O-2, indicating that it has clear O-2 tolerance., INT UNION CRYSTALLOGRAPHY
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, 2016年01月, ［査読有り］

Structural differences of oxidized iron-sulfur and nickel-iron cofactors in O-2-tolerant and O-2-sensitive hydrogenases studied by X-ray absorption spectroscopy
Kajsa G. V. Sigfridsson; Nils Leidel; Oliver Sanganas; Petko Chemev; Oliver Lenz; Ki-Seok Yoon; Hirofumi Nishihara; Alison Parkin; Fraser A. Armstrong; Sebastien Dementin; Marc Rousset; Antonio L. De lacey; Michael Haumann, The class of [NiFe]-hydrogenases comprises oxygen-sensitive periplasmic (PH) and oxygen-tolerant membrane-bound (MBH) enzymes. For three PHs and four MBHs from six bacterial species, structural features of the nickel-iron active site of hydrogen turnover and of the iron-sulfur clusters functioning in electron transfer were determined using X-ray absorption spectroscopy (XAS). Fe-XAS indicated surplus oxidized iron and a lower number of similar to 2.7 angstrom Fe-Fe distances plus additional shorter and longer distances in the oxidized MBHs compared to the oxidized PHs. This supported a double-oxidized and modified proximal FeS cluster in all MBHs with an apparent trimer-plus-monomer arrangement of its four iron atoms, in agreement with crystal data showing a [4Fe3S] cluster instead of a [4Fe4S] cubane as in the PHs. Ni-XAS indicated coordination of the nickel by the thiol group sulfurs of four conserved cysteines and at least one iron-oxygen bond in both MBH and PH proteins. Structural differences of the oxidized inactive [NiFe] cofactor of MBHs in the Ni-B state compared to PHs in the Ni-A state included a similar to 0.05 angstrom longer Ni-O bond, a two times larger spread of the Ni-S bond lengths, and a similar to 0.1 angstrom shorter Ni-Fe distance. The modified proximal [4Fe3S] cluster, weaker binding of the Ni-Fe bridging oxygen species, and an altered localization of reduced oxygen species at the active site may each contribute to O-2 tolerance. (C) 2014 Elsevier B.V. All rights reserved., ELSEVIER SCIENCE BV
BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS, 2015年02月, ［査読有り］

Crystallization and preliminary X-ray analysis of the NAD(+)-reducing [NiFe] hydrogenase from Hydrogenophilus thermoluteolus TH-1
Midori Taketa; Hanae Nakagawa; Mao Habukawa; Hisao Osuka; Kiyohito Kihira; Hirofumi Komori; Naoki Shibata; Masaharu Ishii; Yasuo Igarashi; Hirofumi Nishihara; Ki-Seok Yoon; Seiji Ogo; Yasuhito Shomura; Yoshiki Higuchi, NAD(+)-reducing [NiFe] hydrogenases catalyze the oxidoreduction of dihydrogen concomitant with the interconversion of NAD(+) and NADH. Here, the isolation, purification and crystallization of the NAD(+)-reducing [NiFe] hydrogenase from Hydrogenophilus thermoluteolus TH-1 are reported. Crystals of the NAD(+)-reducing [NiFe] hydrogenase were obtained within one week from a solution containing polyethylene glycol using the sitting-drop vapour-diffusion method and micro-seeding. The crystal diffracted to 2.58 angstrom resolution and belonged to space group C2, with unit-cell parameters a = 131.43, b = 189.71, c = 124.59 angstrom, = 109.42 degrees. Assuming the presence of two NAD(+)-reducing [NiFe] hydrogenase molecules in the asymmetric unit, V-M was calculated to be 2.2 angstrom(3)Da(-1), which corresponds to a solvent content of 43%. Initial phases were determined by the single-wavelength anomalous dispersion method using the anomalous signal from the Fe atoms., WILEY-BLACKWELL
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, 2015年01月, ［査読有り］

Kinetic analysis of inactivation and enzyme reaction of oxygen-tolerant [NiFe]-hydrogenase at direct electron transfer-type bioanode
So; K.; Kitazumi; Y.; Shirai; O.; Kurita; K.; Nishihara; H.; Higuchi; Y.; Kano; K., Membrane-bound [[NiFe]-hydrogenase (MBH) from Hydrogenovibrio marinus is an O-2-tolerant enzyme and allows direct-electron-transfer (DET) bioelectrocatalysis for H-2-oxidation. MBH is a promising bioelectrocatalyst for bioanode of enzymatic H-2-O-2 biofuel cells. From the practical viewpoint of electricity production, the H-2-depletion near the electrode surface and the oxidative and reversible inactivation called "anaerobic inactivation" of [NiFe]-hydrogenases limit the H-2-oxidation at high potentials. We have already proposed a gas-diffusion system to avoid inactivation in our previous study. In this research, we have analyzed the kinetics of the electrochemically induced anaerobic inactivation and the DET bioelectrocatalytic reaction of MBH on electrodes. When the inactivation is considered as a competitive inhibition-like reaction, the maximum value of the apparent Michaelis constant reaches 6.5 mM (at Ketjen Black-modified electrode) as analyzed in our kinetic model. Since the value is larger than the saturated H-2-concentration in solution (0.74 mM), we conclude that high-speed H-2-supply realized by a gas-diffusion electrode is essential to compete with the inactivation. Furthermore, a gas-diffusion bioanode with MBH can eliminate the H-2-depletion near the electrode surface and has reached about 10 mA cm(-2) at 0 V (vs. Ag vertical bar AgCl vertical bar sat. KCl electrode) under quiescent (passive) and H-2-atmospheric conditions., CHEMICAL SOC JAPAN
Bull. Chem. Soc. Jpn., 2014年11月, ［査読有り］

Gas-diffusion and Direct-electron-transfer-type Bioanode for Hydrogen Oxidation with Oxygen-tolerant [NiFe]-hydrogenase as an Electrocatalyst
Keisei So; Yuki Kitazumi; Osamu Shirai; Kouhei Kurita; Hirofumi Nishihara; Yoshiki Higuchi; Kenji Kano, A membrane-bound [NiFe]-hydrogenase (MBH) from Hydrogenovibrio marinus allows the direct-electron-transfer (DET)-type bioelectrocatalysis for H-2 oxidation and is an O-2-tolerant promising enzyme for the construction of enzymatic H-2-O-2 biofuel cells. However, the oxidative and reversible inactivation called "anaerobic inactivation" limits the H-2 oxidation at high potentials. In order to avoid this inhibition, we have constructed an MBH-adsorbed gas-diffusion-type electrode, in which H-2 is spontaneously supplied to the electrode from the gas phase., CHEMICAL SOC JAPAN
CHEMISTRY LETTERS, 2014年10月, ［査読有り］

High thermal stability and unique trimer formation of cytochrome c’ from thermophilic Hydrogenophilus thermoluteolus
Fujii, S; Masanari, M; Inoue, H; Yamanaka, M; Wakai, S; Nishihara, H; Sambongi, Y, Sequence analysis indicated that thermophilic Hydrogenophilus thermoluteolus cytochrome c' (PHCP) and its mesophilic homolog, Allochromatium vinosum cytochrome c' (AVCP), closely resemble each other in a phylogenetic tree of the cytochrome c' family, with 55% sequence identity. The denaturation temperature of PHCP was 87 degrees C, 35 degrees C higher than that of AVCP. Furthermore, PHCP exhibited a larger enthalpy change value during its thermal denaturation than AVCP. While AVCP was dimeric, as observed previously, PHCP was trimeric, and this was the first observation as a cytochrome c'. Dissociation of trimeric PHCP and its protein denaturation reversibly occurred at the same time in a two-state transition manner. Therefore, PHCP is enthalpically more stable than AVCP, perhaps due to its unique trimeric form, in addition to the lower number of Gly residues in its putative at-helical regions., TAYLOR & FRANCIS LTD
Biosci. Biotechnol. Biochem., 2013年08月, ［査読有り］

Oxidative phosphorylation in a thermophilic, facultative chemoautotroph, Hydrogenophilus thermoluteolus, living prevalently in geothermal niches
Satoshi Wakai; Misa Masanari; Takumi Ikeda; Naho Yamaguchi; Saori Ueshima; Kaori Watanabe; Hirofumi Nishihara; Yoshihiro Sambongi, Hydrogenophilus is a thermophilic, facultative chemoautotroph, which lives prevalently in high temperature geothermal niches. Despite the environmental distribution, little is known about its oxidative phosphorylation. Here, we show that inverted membrane vesicles derived from Hydrogenophilus thermoluteolus cells autotrophically cultivated with H2 formed a proton gradient on the addition of succinate, dl-lactate, and NADH, and exhibited oxidation activity toward these three organic compounds. These indicate the capability of mixotrophic growth of this bacterium. Biochemical analysis demonstrated that the same vesicles contained an F-type ATP synthase. The F1 sector of the ATP synthase purified from H. thermoluteolus membranes exhibited optimal ATPase activity at 65°C. Transformed Escherichia coli membranes expressing H. thermoluteolus F-type ATP synthase exhibited the same temperature optimum for the ATPase. These findings shed light on H. thermoluteolus oxidative phosphorylation from the aspects of membrane bioenergetics and ATPase biochemistry, which must be fundamental and advantageous in the biogeochemical cycles occurred in the high temperature geothermal niches. © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.
Environmental Microbiology Reports, 2013年04月, ［査読有り］

Hydrogen-driven asymmetric reduction of hydroxyacetone to (R)-1,2-propanediol by Ralstonia eutropha transformant expressing alcohol dehydrogenase from Kluyveromyces lactis
Takahiro Oda; Koji Oda; Hiroaki Yamamoto; Akinobu Matsuyama; Masaharu Ishii; Yasuo Igarashi; Hirofumi Nishihara, Background: Conversion of industrial processes to more nature-friendly modes is a crucial subject for achieving sustainable development. Utilization of hydrogen-oxidation reactions by hydrogenase as a driving force of bioprocess reaction can be an environmentally ideal method because the reaction creates no pollutants. We expressed NAD-dependent alcohol dehydrogenase from Kluyveromyces lactis in a hydrogen-oxidizing bacterium: Ralstonia eutropha. This is the first report of hydrogen-driven in vivo coupling reaction of the alcohol dehydrogenase and indigenous soluble NAD-reducing hydrogenase. Asymmetric reduction of hydroxyacetone to (R)-1,2-propanediol, which is a commercial building block for antibacterial agents, was performed using the transformant as the microbial cell catalyst.
Results: The two enzymes coupled in vitro in vials without a marked decrease of reactivity during the 20 hr reaction because of the hydrogenase reaction, which generates no by-product that affects enzymes. Alcohol dehydrogenase was expressed functionally in R. eutropha in an activity level equivalent to that of indigenous NAD-reducing hydrogenase under the hydrogenase promoter. The hydrogen-driven in vivo coupling reaction proceeded only by the transformant cell without exogenous addition of a cofactor. The decrease of reaction velocity at higher concentration of hydroxyacetone was markedly reduced by application of an in vivo coupling system. Production of (R)-1,2-propanediol (99.8% e.e.) reached 67.7 g/l in 76 hr with almost a constant rate using a jar fermenter. The reaction velocity under 10% P-H2 was almost equivalent to that under 100% hydrogen, indicating the availability of crude hydrogen gas from various sources. The in vivo coupling system enabled cell-recycling as catalysts.
Conclusions: Asymmetric reduction of hydroxyacetone by a coupling reaction of the two enzymes continued in both in vitro and in vivo systems in the presence of hydrogen. The in vivo reaction system using R. eutropha transformant expressing heterologous alcohol dehydrogenase showed advantages for practical usage relative to the in vitro coupling system. The results suggest a hopeful perspective of the hydrogen-driven bioprocess as an environmentally outstanding method to achieve industrial green innovation. Hydrogen-oxidizing bacteria can be useful hosts for the development of hydrogen-driven microbial cell factories., BIOMED CENTRAL LTD
MICROBIAL CELL FACTORIES, 2013年01月, ［査読有り］

Structural basis for a [4Fe-3S] cluster in the oxygen-tolerant membrane-bound [NiFe]-hydrogenase
Yasuhito Shomura; Ki-Seok Yoon; Hirofumi Nishihara; Yoshiki Higuchi, Membrane-bound respiratory [NiFe]-hydrogenase (MBH), a H-2-uptake enzyme found in the periplasmic space of bacteria, catalyses the oxidation of dihydrogen: H-2 -> 2H(+) + 2e(-) (ref. 1). In contrast to the well-studied O-2-sensitive [NiFe]-hydrogenases (referred to as the standard enzymes), MBH has an O-2-tolerant H-2 oxidation activity(2-4); however, the mechanism of O-2 tolerance is unclear(5). Here we report the crystal structures of Hydrogenovibrio marinus MBH in three different redox conditions at resolutions between 1.18 and 1.32 angstrom. We find that the proximal iron-sulphur (Fe-S) cluster of MBH has a [4Fe-3S] structure coordinated by six cysteine residues-in contrast to the [4Fe-4S] cubane structure coordinated by four cysteine residues found in the proximal Fe-S cluster of the standard enzymes-and that an amide nitrogen of the polypeptide backbone is deprotonated and additionally coordinates the cluster when chemically oxidized, thus stabilizing the superoxidized state of the cluster. The structure of MBH is very similar to that of the O-2-sensitive standard enzymes except for the proximal Fe-S cluster. Our results give a reasonable explanation why the O-2 tolerance of MBH is attributable to the unique proximal Fe-S cluster; we propose that the cluster is not only a component of the electron transfer for the catalytic cycle, but that it also donates two electrons and one proton crucial for the appropriate reduction of O-2 in preventing the formation of an unready, inactive state of the enzyme., NATURE PUBLISHING GROUP
NATURE, 2011年11月, ［査読有り］

Purification and characterization of a highly thermostable, oxygen-resistant, respiratory [NiFe]-hydrogenase from a marine, aerobic hydrogen-oxidizing bacterium Hydrogenovibrio marinus
Ki-Seok Yoon; Keiichi Fukuda; Kiyoshi Fujisawa; Hirofumi Nishihara, The membrane-bound [NiFe]-hydrogenase from Hydrogenooibrio marinus (HmMBH) was purified homogeneously under anaerobic conditions. Its molecular weight was estimated as 110 kDa, consisting of a heterodimeric structure of 66 kDa and 37 kDa subunits. The purified enzyme exhibited high activity in a wide temperature range: 185 U/mg at 30 degrees C and 615 U/mg at 85 degrees C (the optimum temperature). The Km and k(cat)/K(m) values for H(2) were, respectively, 12 mu M and 8.58 x 10(7) M(-1) s(-1). The optimum reaction pH was 7.8, but its stability was particularly high at pH 4.0-7.0. Results show that HmMBH was remarkably thermostable and oxygen-resistant: its half-life was 75 h at 80 degrees C under H(2), and more than 72 h at 4 degrees C under air. The air-oxidized HmMBH for 72 h showed only weak EPR signals of Ni-B, suggesting a structural feature in which the active center is not easily oxidized. Copyright (C) 2011, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved., PERGAMON-ELSEVIER SCIENCE LTD
INTERNATIONAL JOURNAL OF HYDROGEN ENERGY, 2011年06月, ［査読有り］

Crystallization and preliminary X-ray diffraction analysis of membrane-bound respiratory [NiFe] hydrogenase from Hydrogenovibrio marinus.
Shomura; Y.; Hagiya; K.; Yoon; K. S.; Nishihara; H.; Higuchi; Y., Membrane-bound respiratory [NiFe] hydrogenase is an H-2-uptake enzyme found in the periplasmic space of bacteria that plays a crucial role in energy-conservation processes. The heterodimeric unit of the enzyme from Hydrogeno-vibrio marinus was purified to homogeneity using chromatographic procedures. Crystals were grown using the sitting-drop vapour-diffusion method at room temperature. Preliminary crystallographic analysis revealed that the crystals belonged to space group P2(1), with unit-cell parameters a = 75.72, b = 116.59, c = 113.40 angstrom, beta = 91.3 degrees, indicating that two heterodimers were present in the asymmetric unit., WILEY-BLACKWELL
Acta Cryst., 2011年, ［査読有り］

Heterologous synthesis of cytochrome c’ by Escherichia coli is not dependent on the System I cytochrome c biogenesis machinery
Inoue, H; Wakai, S; Nishihara, H; Sambongi, Y, Hydrogenophilus thermoluteolus cytochrome c' (PHCP) has typical spectral properties previously observed for other cytochromes c', which comprise Ambler's class II cytochromes c. The PHCP protein sequence (135 amino acids) deduced from the cloned gene is the most homologous (55% identity) to that of cytochrome c' from Allochromatium vinosum (AVCP). These findings indicate that PHCP forms a four-helix bundle structure, similar to AVCP. Strikingly, PHCP with a covalently bound heme was heterologously synthesized in the periplasm of Escherichia coli strains deficient in the DsbD protein, a component of the System I cytochrome c biogenesis machinery. The heterologous synthesis of PHCP by aerobically growing E. coli also occurred without a plasmid carrying the genes for Ccm proteins, other components of the System I machinery. Unlike Ambler's class I general cytochromes c, the synthesis of PHCP is not dependent on the System I machinery and exhibits similarity to that of E. coli periplasmic cytochrome b(562), a 106-residue four-helix bundle., WILEY-BLACKWELL
FEBS J., 2011年, ［査読有り］

Purification and biochemical characterization of a membrane-bound [NiFe]-hydrogenase from a hydrogen-oxidizing, lithotrophic bacterium, Hydrogenophaga sp AH-24
Ki-Seok Yoon; Yukiko Sakai; Natsuki Tsukada; Kiyoshi Fujisawa; Hirofumi Nishihara, Membrane-bound [NiFe]-hydrogenase from Hydrogenophaga sp. AH-24 was purified to homogeneity. The molecular weight was estimated as 100 +/- 10 kDa, consisting of two different subunits (62 and 37 kDa). The optimal pH values for H-2 oxidation and evolution were 8.0 and 4.0, respectively, and the activity ratio (H-2 oxidation/H-2 evolution) was 1.61 x 10(2) at pH 7.0. The optimal temperature was 75 degrees C. The enzyme was quite stable under air atmosphere (the half-life of activity was c. 48 h at 4 degrees C), which should be important to function in the aerobic habitat of the strain. The enzyme showed high thermal stability under anaerobic conditions, which retained full activity for over 5 h at 50 degrees C. The activity increased up to 2.5-fold during incubation at 50 degrees C under H-2. Using methylene blue as an electron acceptor, the kinetic constants of the purified membrane-bound homogenase (MBH) were V-max=336 U mg(-1), k(cat)=560 s(-1), and k(cat)/K-m=2.24 x 10(7) M-1 s(-1). The MBH exhibited prominent electron paramagnetic resonance signals originating from [3Fe-4S](+) and [4Fe-4S](+) clusters. On the other hand, signals originating from Ni of the active center were very weak, as observed in other oxygen-stable hydrogenases from aerobic H-2-oxidizing bacteria. This is the first report of catalytic and biochemical characterization of the respiratory MBH from Hydrogenophaga., OXFORD UNIV PRESS
FEMS MICROBIOLOGY LETTERS, 2009年01月, ［査読有り］

Isolation and characterization of a new facultatively autotrophic hydrogen-oxidizing Betaproteobacterium, Hydrogenophaga sp. AH-24.
Yoon; K. S.; Tsukada; N.; Sakai; Y.; Ishii; M.; Igarashi; Y.; Nishihara; H., 責任著者, A hydrogen-oxidizing bacterium strain AH-24 was isolated, which was classified in the genus Hydrogenophaga, based on the 16S rRNA gene sequence. The isolate possessed a typical yellow pigment of Hydrogenophaga species. Its closest relative was Hydrogenophaga pseudoflava, but the assimilation profile of sugar compounds resembled that of no species of Hydrogenophaga. The optimum temperature and pH for autotrophic growth were, respectively, 33-35 degrees C and 7.0. Most hydrogenase activity (benzyl viologen reducing activity) was localized in the membrane fraction (MF), but NAD(P)-reducing hydrogenase activity was detected in neither the membrane nor the soluble fractions. Cytochromes b(561) and c(551) were present in MF; both were reduced when hydrogen was supplied to the oxidized MF, suggesting involvement in respiratory H-2 oxidation as electron carriers. Cytochrome b(561) was inferred to function as the redox partner of the membrane-bound hydrogenase., OXFORD UNIV PRESS
FEMS Microbiol. Lett., 2008年, ［査読有り］

Thiosulfate oxidation by a moderately thermophilic hydrogen-oxidizing bacterium, Hydrogenophilus thermoluteolus
Daisuke Miyake; Shin-ichi Ichiki; Miyako Tanabe; Takahiro Oda; Hisao Kuroda; Hirofumi Nishihara; Yoshihiro Sambongi, The moderately thermophilic Betaproteobacterium, Hydrogenophilus thermoluteolus, not only oxidizes hydrogen, the principal electron donor for growth, but also sulfur compounds including thiosulfate, a process enabled by sox genes. A periplasmic extract of H. thermoluteolus showed significant thiosulfate oxidation activity. Ten genes apparently involved in thiosulfate oxidation (soxEFCDYZAXBH) were found on a 9.7-kb DNA fragment of the H. thermoluteolus chromosome. The proteins SoxAX, which represent c-type cytochromes, were co-purified from the cells of H. thermoluteolus; they enhanced the thiosulfate oxidation activity of the periplasmic extract when added to the latter., SPRINGER
ARCHIVES OF MICROBIOLOGY, 2007年08月, ［査読有り］

Cupriavidus pinatubonensis sp. nov. and Cupriavidus laharis sp. nov., novel hydrogen-oxidizing, facultatively chemolithotrophic bacteria isolated from volcanic mudflow deposits from Mt. Pinatubo in the Philippines.
Sato; Y.; Nishihara; H.; Yoshida; M.; Watanabe; M.; Rondal; J. D.; Concepcion; R. N.; Ohta; H., Taxonomic studies were performed on ten hydrogen-oxidizing, facultatively chemolithotrophic bacteria that were isolated from volcanic mudflow deposits derived from the eruption of Mt. Pinatubo in the Philippines in 1991. Phylogenetic analysis based on 16S rRNA gene sequences indicated that these isolates belonged to the genus Cupriavidus of the Betaproteobacteria; sequence similarity values with their nearest phylogenetic neighbour, Cupriavidus basilensis, were 97(.)1-98(.)3 %. In addition to phylogenetic analysis, results of whole-cell protein profiles and biochemical tests revealed that these strains were members of two distinct species. DNA-DNA hybridizations and whole-cell protein profiles enabled these isolates to be differentiated from related Cupriavidus species with validly published names. The isolates were aerobic, Gramnegative, non-sporulating, peritrichously flagellated rods. Their G+C contents ranged from 65(.)2 to 65(.)9 mol% and their major isoprenoid quinone was ubiquinone Q-8. On the basis of these results, two novel species are proposed, Cupriavidus pinatubonensis sp. nov. [nine strains, with 1245(T) (=CIP 108725(T)= PNCM 10346(T)) as the type strain] and Cupriavidus laharis sp. nov. [one strain, the type strain 1263a(T) (=CIP 108726(T)= PNCM 10347(T))]. It is also suggested that Ralstonia sp. LMG 1197 (=JMP 134) should be included in the species C. pinatubonensis., SOC GENERAL MICROBIOLOGY
Int. J. Syst. Evol. Microbiol., 2006年, ［査読有り］

Light-driven hydrogen production by a hybrid complex of a [NiFe]-hydrogenase and the cyanobacterial photosystem I
Ihara; M.; Nishihara; H.; Yoon; K. S.; Lenz; O.; Friedrich; B.; Nakamoto; H.; Kojima; K.; Honma; D.; Kamachi; T.; Okura; I., In order to generate renewable and clean fuels, increasing efforts are focused on the exploitation of photosynthetic microorganisms for the production of molecular hydrogen from water and light. In this study we engineered a 'hard-wired' protein complex consisting of 9 hydrogenase and photosystem I (hydrogenase-PSI complex) as a direct light-to-hydrogen conversion system. The key component was an artificial fusion protein composed of the membrane-bound [NiFe] hydrogenase from the beta-proteobacterium Ralstonia eutropha H16 and the peripheral PSI subunit PsaE of the cyanobacterium Thermosy-nechococcus elongatus. The resulting hydrogenase-PsaE fusion protein associated with PsaE-free PSI spontaneously, thereby forming a hydrogenase-PSI complex as confirmed by sucrose-gradient ultracentrifuge and immunoblot analysis. The hydrogenase-PSI complex displayed light-driven hydrogen production at a rate of 0.58 mu mol H-2 center dot mg chlorophyll(-1)center dot h(-1). The complex maintained its accessibility to the native electron acceptor ferredoxin. This study provides the first example of a light-driven enzymatic reaction by an artificial complex between a redox enzyme and photosystem I and represents an important step on the way to design a photosynthetic organism that efficiently converts solar energy and water into hydrogen., AMER SOC PHOTOBIOLOGY
Photochem. Photobiol., 2006年, ［査読有り］

Cloning, expression, crystallization and preliminary X-ray characterization of cytochrome c552 from a moderate thermophilic bacterium, Hydrogenophilus thermoluteolus.
Ichiki; S.; Nakamura; S.; Ohkubo; T.; Kobayashi; Y.; Hasegawa; J.; Uchiyama; S.; Nishihara; H.; Mizuta; K.; Sambongi; Y., BLACKWELL PUBLISHING
Acta Cryst., 2005年, ［査読有り］

Occurrence of hydrogen-oxidizing Ralstonia species as primary microorganisms in the Mt. Pinatubo volcanic mudflow deposits
Sato; Y.; Nishihara; H.; Yoshida; M.; Watanabe; M.; Rondal; J.D. and Ohta; H., Ralstonia eutropha is an aerobic, hydrogen-oxidizing, facultative chemolithotrophic bacterium. Recently, R. eutropha-related bacteria have been found to predominate in the cultural bacterial communities of fresh mudflow deposits from Mt. Pinatubo, the Philippines. In the present study, the R. eutropha-related strains were examined for their chemolithotrophic growth in the presence of H-2, O-2, and CO2 (85:5:10) and for their phylogenetic relationship with R. eutropha. Eleven of the 12 strains tested grew in the presence of H-2, O-2, and CO2, although the growth was not as rapid as that of the well-characterized H-2-oxidizing strain, R. eutropha H16. To examine the hydrogenase activity, membrane and soluble fractions were prepared from R. eutropha-related strain 1245 and its activity was assayed with methylene blue and NAD as electron acceptors. A significant hydrogenase activity was detected in both membrane and soluble fractions of the cell-free extract. The 16S rDNA analysis of the 12 strains of R. eutropha-related bacteria revealed that they could be classified into two clusters, both of which were clearly separated from the cluster of R. eutropha sensu stricto. The probable ecological niche of H2-oxidizing Ralstonia species in the volcanic mudflow deposits was examined., JAPANESE SOC SOIL SCIENCE PLANT NUTRITION
Soil Sci. Plant Nutr., 2004年, ［査読有り］

Quantitative and rapid detection of the trichloroethylene-degrading bacterium Methylocystis sp. M in groundwater by real-time PCR
Kikuchi; T.; Iwasaki; K.; Nishihara; H.; Takamura; Y.; Yagi; O., We developed a method based on real-time PCR for the specific and rapid enumeration of a trichloroethylene-degrading methanotroph, Methylocystis sp. M, with the aim of monitoring the strain in groundwater. A primer set designed from the nucleotide sequence of the mmoC gene of a soluble methane monooxygenase (sMMO) gene cluster from Methylocystis sp. M was specific to amplify the DNA region from the strain and no PCR products were amplified with the sMMO gene clusters from six other methanotroph strains. The real-time PCR reliably quantified Methylocystis sp. M over at least five orders of magnitude (5x10(6) to 5x10(2) cells/PCR tube, or 2x10(8) to 2x10(4) cells/ml). Five cells of Methylocystis sp. M per PCR tube (2x10(2) cells/ml) were detectable when the cells were suspended in distilled water. The concomitant presence of other methanotrophs in samples did not affect the reliability of enumeration; and recovery of the cells with a membrane filter enabled us to quantify cells of the strain in groundwater. This quantification procedure was completed within 3 h, including preparation time of environmental samples. We conclude that real-time PCR using the mmoC primer set can be used practically to analyze the behavior of Methylocystis sp. M at bioremediation sites., SPRINGER-VERLAG
Appl. Microbiol. Biotechnol., 2002年, ［査読有り］

Analysis of the molecular species of hydrogenase in the cells of an obligately chemolithoautotrophic, marine hydrogen-oxidizing bacterium, Hydrogenovibrio marinus
Nishihara; H.; Miyata; Y.; Miyashita; Y. Bernhard; M.; Pohlmann; A.; Friedrich; B. Takamura; Y., 筆頭著者, Hydrogenovibrio marinus was suggested to have only membrane-bound hydrogenase (MBH). The change of cultivation pO(2) did not affect the molecular species of hydrogenase expressed. We propose the MBH is grouped in class I [NiFe] MBH according to the subunit composition, size (Mw 38,000 and Mw 74,000 subunits) and N-terminal sequences of the subunits, and arrangement of the structural genes. Ni-requirement for the autotrophic growth on H-2 also suggested the MBH is the Ni-containing type. Southern hybridization analysis using a part of the MBH gene showed a possibility of the presence of two highly homologous MBHs which were not separated by SDS-PAGE., TAYLOR & FRANCIS LTD
Biosci. Biotechnol. Biochem., 2001年, ［査読有り］

Quantitative and specific detection of a trichloroethylene-degrading methanotroph, Methylocystis sp. strain M, by a most probable number-polymerase chain reaction method
Kikuchi; T.; Iwasaki; K.; Nishihara; H.; Takamura; Y.; Yagi; O., We developed a rapid and specific enumeration method for a trichloroethylene-degrading methanotroph. Methylocystis sp. strain M, based on a most probable number-polymerase chain reaction method for monitoring the bacterium at bioremediation sites. The primers designed for the mmoC gene of the soluble methane monooxygenase gene cluster were specific to strain M. Recovery of the cells with a membrane filter enabled us to detect strain M in trichloroethylene-contaminated groundwater. We used the enumeration method to monitor the number of strain M cells in effluent from soil columns supplied with trichloroethylene-contaminated groundwater. The number of strain M cells in the effluent depended on the amount of the strain M inoculated and the number of cells measured by the most probable number-polymerase chain reaction method was correlated with that measured by a culture method. The detection limit for strain M in effluent detected by MPN-PCR method was 4 to 8 x 10(2) cells/ml., TAYLOR & FRANCIS LTD
Biosci. Biotechnol. Biochem., 2001年, ［査読有り］

Sequences and gene organization of genes of an extremely thermophilic and oxygen-stable membrane-bound hydrogenase from Hydrogenovibrio marinus　　　　　　　　　　　　　　　
Nishihara, H; Bernhard, M; Pohlmann, A; Takamura, Y; Friedrich B, 筆頭著者
Microbiology 2000-Health, Ecology, Biotechnology, 2000年, ［査読有り］

Microcystis 属および Oscillatoria 属 シアノバクラリアの各種有機物に対する増殖応答とL-リジンとL-ヒスチジンによる増殖阻害
高村義親; 千野まりこ; 長田綾子; 西原宏史; 矢木修身, 環境科学会
環境科学会誌, 1999年, ［査読有り］

Oxidation of uronic acids by a large excess of glucose oxidase preparations　　　　　　　　　　　　　　　
Kobayashi, M; Nishihara, H; Kobayashi, S
Journal of Applied Glycosciences, 1999年, ［査読有り］

Acidic polysaccharide production from uronic acids by facultative anaerobe of Aeromonas sp. GLCA 9B　　　　　　　　　　　　　　　
Fukami, K; Nishihara, H; Takamura, Y; Ishii, T; Funane, K; Kitamura, Y; Kobayashi, M
Journal of Applied Glycosciences, 1999年, ［査読有り］

Cloning and characterization of mdc genes encoding malonate decarboxylase from Pseudomonas putida
Chohnan; S.; Kurusu; Y.; Nishihara; H.; Takamura; Y., The DNA fragment encoding malonate decarboxylase, involved in malonate assimilation, was cloned from Pseudomonas putida. The Il-kb DNA fragment contained nine open reading frames, which were designated mdcABCDEGHLM in the given order. N-terminal protein sequencing established that the mdcA, mdcC, mdcD, mdcE and mdcH genes encoded subunits alpha, delta, beta, gamma and epsilon Of the malonate decarboxylase, respectively. Malonate decarboxylase was functionally expressed in Escherichia coli from plasmid harboring the entire gene cluster or the mdc genes lacking the mdcL and mdcM genes. The mdcL and mdcM genes encode membrane proteins and disruption of the genes of P. putida by the insertion of a kanamycin resistance cassette reduced the malonate uptake activity of the organism. Thus, we conclude that MdcLM is a malonate transporter. (C) 1999 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved., ELSEVIER SCIENCE BV
FEMS Microbiology Letters, 1999年, ［査読有り］

Phylogenetic position of an obligately chemoautotrophic, marine hydrogen-oxidizing bacterium, Hydrogenovibrio marinus, on the basis of 16s rRNA gene sequences and two form I RuBisco gene sequences
Nishihara; H.; Yaguchi; T.; Chung; S. Y.; Suzuki; K.; Yanagi; M.; Yamasato; K.; Kodama; T.; Igarashi; Y., 筆頭著者, Two form ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) genes from the obligately autotrophic, marine hydrogen oxidizer Hydrogenovibrio marinus were sequenced. The deduced amino acid sequences of both RuBisCOs revealed that they are similar to those of sulfur oxidizers (Thiobacillus) and a purple sulfur bacterium (Chromatium vinosum). According to the 16S rRNA gene sequences, H. marinus is also affiliated with these microorganisms, members of Thiomicrospira being the closest relatives. Sequence similarities of the 16S rRNA genes and of the RuBisCO genes among these gamma-Proteobacteria suggest a common autotrophic ancestry. An ancestor of purple sulfur bacteria might be a common root of H. marinus and related sulfur oxidizers., SPRINGER VERLAG
Archives of Microbiology, 1998年, ［査読有り］

Different properties of gene products of three sets of Ribulose 1,5-bisphosphate carboxylase/oxygenase from a marine obligately autotrophic hydrogen-oxidizing bacterium, Hydrogenovibrio marinus strain MH-110
Hayashi; N. R.; Oguni; A.; Yaguchi; T.; Chung; S. Y.; Nishihara; H.; Kodama; T.; Igarashi; Y., Hydrogenovibrio marinas strain MH-110 is an obligately lithoautotrophic hydrogen-oxidizing bacterium, possessing three sets of the genes for ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO); one form II type (L-x) and two form I type (L8S8) enzymes. The genes for the form I type enzymes are named cbbLS-1 and cbbLS-2, and that for the form II type enzyme is named cbbM. These three sets of genes were cloned in plasmid vectors, and the expression of the genes in E. coli cells were studied. The three RubisCOs were purified through the same steps, and their specificity factors (tau values) were measured. The tau values of CbbLS-1 and CbbLS-2 were higher than that of CbbM, but lower than those of other form I RubisCOs. The specific activity of CbbM was very low and CbbM was inactivated easily during the process of purification. Immunochemical analyses revealed that the antibody against form I type reacted only with CbbL-1 and CbbL-2, and the antibody against form II type reacted only with CbbM. The antibody to neither form of RubisCO demonstrated cross-reactivity for the other form., SOC FERMENTATION BIOENGINEERING, JAPAN
Journal of Fermentation and Bioengineering, 1998年, ［査読有り］

Malonate decarboxylase of Pseudomonas putida is composed of five subunits
Chohnan; S.; Fujio; T.; Takaki; T.; Yonekura; M.; Nishihara; H.; Takamura; Y., Two different forms of malonate decarboxylase were purified from Pseudomonas putida. The active form was composed of the five different subunits alpha (60 kDa), beta (33 kDa), gamma (28 kDa), delta (13 kDa), and epsilon (30 kDa) and the inactive form was composed of the four subunits lacking the epsilon subunit. The former catalyzed the decarboxylation of malonate to acetate, but the latter could not, although it retained both activities of acetyl-CoA:malonate CoA transferase and malonyl-CoA decarboxylase. The delta subunit of the active form was acylated by the incubation with [2-C-14]malonyl-CoA, but the delta subunit of the inactive form was not labeled. From the above results and the N-terminal amino acid sequence analysis, it was concluded that the epsilon subunit was an essential subunit to function as malonyl-CoA:ACP transacylase, which was an indispensable component of the enzyme for the cyclic decarboxylation of malonate. (C) 1998 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved., ELSEVIER SCIENCE BV
FEMS Microbiology Letters, 1998年, ［査読有り］

Changes in size of intracellular pools of coenzyme A and its thioesters in Escherichia coli K-12 cells to various carbon sources and stresses
Chohnan; S.; Izawa; H.; Nishihara; H.; Takamura; Y., Intracellular pools of three CoA molecular species of coenzyme A, CoASH, acetyl-CoA, and malonyl-CoA, in Escherichia coli K-12 cells were studied by acyl-CoA cycling method in replacement culture. The sizes and compositions of CoA pools starved for a carbon source changed within minutes after the addition of one of various carbon sources. A large acetyl-CoA pool formed after the addition of D-glucose, D-fructose, D-mannose, glycerol, or sorbitol, but there was little change when L-glucose, sucrose, maltose, succinate, or acetate was added, The beta-anomer of D-glucose was assimilated 10 times faster than the alpha-anomer, Intracellular CoA pools also changed with stress: in the pH, incubation temperature, or with osmotic stress. The sizes and compositions of CoA pools were not affected by pH changing between 4 and 8, but the breakdown of acetyl-CoA and CoASH was greater at pH 9 than at pH 4 to 8. Production of acetyl-CoA was greatest at 40 degrees C, and at 50 degrees C, an acetyl-CoA pool did not form at all and the size of the CoASH pool declined. When the organism was stressed by the addition of NaCl at concentrations of more than 0.6 M, little acetyl-CoA was produced. The total CoA pool (the sum of the concentrations of CoASH, acetyl-CoA, and malonyl-CoA) remained within the limits of 0.83-1.40 nmol/mg of dry cell weight (0.30-0.52 mM). Whenever acetyl-CoA increased, CoASH decreased. Therefore, the acetyl-CoA/CoASB ratio is an important index of facultative anaerobes that reflects the state of carbon and energy metabolism in vivo., TAYLOR & FRANCIS LTD
Biosci.Biotechnol. Biochem., 1998年, ［査読有り］

Characterization of an extremely thermophilic and oxygen-stable membrane-bound hydrogenase from a marine hydrogen-oxidizing bacterium Hydrogenovibrio marinus
Nishihara; H.; Miyashita; Y.; Aoyama; K.; Kodama; T.; Igarashi; Y.; Takamura; Y., 筆頭著者, The membrane-bound hydrogenase from a marine hydrogen-oxidizing bacterium Hydrogenovibrio marinus was characterized as highly oxygen-tolerant, extremely thermophilic and thermostable in its membrane-bound form. The optimum temperatures for H-2 oxidation and H-2 evolution were 90 and 80 degrees C, respectively. The enzyme retained 90% of its activity after heating at 70 degrees C for 50 min under air and retained full activity at 90 degrees C for 80 min under hydrogen. The optimum pH values were 5.5 (H-2 evolution) and 9.4 (H-2 oxidation), and the activity ratio (H-2 evolution/H-2 oxidation) was high (50.3) at pH 5.5. The hydrogen evolution from reduced methyl viologen continued at high temperatures and acidic pH with the continuous addition of sodium dithionite. (C) 1997 Academic Press., ACADEMIC PRESS INC ELSEVIER SCIENCE
Biochemical and Biophysical Research Communications, 1997年, ［査読有り］

Random amplified polymorphic DNA (RAPD) analysis for discriminating genotypes of Microcystis cyanobacteria
Nishihara; H.; Miwa; H.; Watanabe; M.; Nagashima; M.; Yagi; O.; Takamura; Y., 筆頭著者, Random amplified polymorphic DNA (RAPD) analysis was used to discriminate genotypes in five species of Microcystis cyanobacteria, Strains of each group with the identical allozyme genotype (T. Kato et al,, Algol, Stud., 1991, 129-140; M. Watanabe, in ''Toxic Microcystis,'' ed. by M. F. Watanabe et al., CRC Press, Tokyo, 1966, pp, 13-34) gave similar RAPD patterns characterizing the respective group, On the other hand, no similarities in RAPD patterns were observed among strains of which allozyme genotypes were different, A good accordance between the RAPD analysis and allozyme divergence indicated a high reliability of both methods for discrimination of the affiliated groups of Microcystis. Several amplified DNA fragments, which were expected to be markers for a particular taxon with identical allozyme genotype, were also observed on the RAPD patterns, Genetic homogeneities of M. novacekii, M. viridis, and M. wesenbergii were shown by RAPD analysis as well as the allozyme genotype, However, significant variations were observed in M. aeruginosa and M. ichthyoblabe in the levels of DNA and proteins (allozymes)., TAYLOR & FRANCIS LTD
Bioscience, Biotechnology and Biochemistry, 1997年, ［査読有り］

An extremely thermophilic and oxygen-stable membrane-bound hydrogenase from a marine hydrogen-oxidizing bacterium Hydrogenovibrio marinus　　　　　　　　　　　　　　　
Nishihara, H; Miyashita, Y; Aoyama, K; Miyata, Y; Kodama, T; Igarashi, Y; Takamura, Y, 筆頭著者
5th International Conference on the Molecular Biology of Hydrogenase, Abstract book, 1997年, ［査読有り］

Changes in the size and composition of intracellular pools of nonesterified coenzyme A and coenzyme A thioesters in aerobic and facultatively anaerobic bacteria
Chohnan; S.; Furukawa; H.; Fujio; T.; Nishihara; H.; Takamura; Y., Intracellular levels of three coenzyme A (CoA) molecular species, i.e., nonesterified CoA (CoASH), acetyl-CoA, and malonyl-CoA, in a variety of aerobic and facultatively anaerobic bacteria were analyzed by the acyl-CoA cycling method developed by us, It was demonstrated that there was an intrinsic difference between aerobes and facultative anaerobes in the changes in the size and composition of CoA pools, The CoA pools in the aerobic bacteria hardly changed and were significantly smaller than those of the facultatively anaerobic bacteria, On the other hand, in the facultatively anaerobic bacteria, the size and composition of the CoA pool drastically changed within minutes in response to the carbon and energy source provided, Acetyl-CoA was the major component of the CoA pool in the facultative anaerobes grown on sufficient glucose, although CoASH was dominant in the aerobes, Therefore, the acetyl-CoA/CoASH ratios in facultatively anaerobic bacteria were 10 times higher than those in aerobic bacteria, In Escherichia coli K-12 cells, the addition of reagents to inhibit the respiratory system led to a rapid decrease in the amount of acetyl-CoA with a concomitant increase in the amount of CoASH, whereas the addition of cerulenin, a specific inhibitor of fatty acid synthase, triggered the intracellular accumulation of malonyl-CoA. The acylation and deacylation of the three CoA molecular species coordinated with the energy-yielding systems and the restriction of the fatty acid-synthesizing system of cells, These data suggest that neither the accumulation of acetyl-CoA nor that of malonyl-CoA exerts negative feedback on pyruvate dehydrogenase and acetyl-CoA carboxylase, respectively., AMER SOC MICROBIOLOGY
Applied and Environmental Microbiology, 1997年, ［査読有り］

Isolation and cultivation of thermophilic cyanobacteria from hot springs of Northern Thailand
Hayashi; N.; Peerapornpisal; Y.; Nishihara; H.; Ishii; M.; Igarashi; Y.; Kodama; T., Three cyanobacterial strains were obtained from hot springs of Northern Thailand. Morphological and physiological studies revealed that all three strains belong to genus Chroococcidiopsis. One of these strains, TS-821, was used for further studies because of its stable autotrophic growth. Its optimum temperature for growth is around 50 degrees C, and its optimum pH for growth is around 8. The strain is capable of dinitrogen fixation, and is tolerant against nitrite (5 mM), sulfite (0.6 mM) and a high concentration of carbon dioxide in the gas phase (0.4 atm). These properties indicate that the strain can be used for the conversion to organic matter of carbon dioxide produced by combustion., SOC FERMENTATION BIOENGINEERING, JAPAN
Journal of Fermentation and Bioengineering, 1994年, ［査読有り］

Purification of form L₂ RubisCO from a marine obligately autotrophic hydrogen-oxidizing bacterium, Hydrogenovibrio marinus strain MH-110
Chung; S.; Yaguchi; T.; Nishihara; H.; Igarashi; Y.; Kodama; T., Ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) was purified from an obligately autotrophic hydrogen-oxidizing bacterium, Hydrogenovibrio marinus MH-110. The protein has a M(r) value of approximately 110000, and is composed of two identical subunits of 55 000. To our knowledge, the existence of L2-form RubisCO in a chemolithoautotrophic bacterium is first reported in this paper. The N-terminal amino acid sequence determination of the purified enzyme showed high homology with those of the L2-form RubisCO of Rhodospirillum rubrum and the L(x)-form RubisCO from Rhodobacter sphaeroides., OXFORD UNIV PRESS
FEMS Microbiology Letters, 1993年, ［査読有り］

Production and properties of glycogen in the marine obligate chemolithoautotroph, Hydrogenovibrio marinus
Nishihara; H.; Igarashi; Y.; Kodama; T.; Nakajima; T., 筆頭著者, Production of an intracellular polysaccharide was observed during cultivation of a marine, obligately chemolithoautotrophic hydrogen-oxidizing bacterium, Hydrogenovibrio marinus, under nitrogen or magnesium starvation or oxygen limitation in a continuous gas supply. The polysaccharide formed in oxygen-limited cells was isolated and shown to be composed of only glucose. Instrumental and enzymatic analyses revealed that the polysaccharide has a glycogen-like structure., SOC FERMENTATION BIOENGINEERING, JAPAN
Journal of Fermentation and Bioengineering, 1993年, ［査読有り］

Biomass production from carbon dioxide by a marine hydrogen-oxidizing bacterium Hydrogenovibrio marinus　　　　　　　　　　　　　　　
Igarashi, Y; Nishihara, H; Kodama, T
Asia-Pacific Biochemical Engineering Conference '92, Abstract book, 1992年, ［査読有り］

RuBisCOs from an obligately chemolithoautotrophic, marine hydrogen-oxidizing bacterium, Hydrogenovibrio marinus　　　　　　　　　　　　　　　
Igarashi, Y; Chung, S; Nishihara, H; Kodama, T
7th International Symposium on Microbial growth on C1 compounds, 1992年, ［査読有り］

Hydrogenovibrio marinus gen. nov. , sp. nov. , a marine obligately chemolithoautotrophic hydrogen-oxidizing bacterium
Nishihara; H.; Igarashi; Y.; Kodama; T., 筆頭著者, The name Hydrogenovibrio marines gen. nov. sp. nov. is proposed for an obligately chemolithoautotrophic, mesophilic, gram-negative, motile, comma-shaped, aerobic, hydrogen-oxidizing bacterium that was isolated from seawater. The optimum temperature and NaCl concentration for growth are 37-degrees-C and 0.5 M, respectively. The guanine-plus-cytosine content of the DNA is 44.1 mol%. The ubiquinone is ubiquinone-8, and the major cellular fatty acids are C16:0, C18:0, and C16:1 acids. The type strain of this species is strain MH-110 ( = JCM 7688)., AMER SOC MICROBIOLOGY
International Journal of Systematic Bacteriology, 1991年, ［査読有り］

Growth characteristics and high cell-density cultivation of a marine obligately chemolithoautotrophic hydrogen-oxidizing bacterium Hydrogenovibrio marinus strain MH-110 under a continuous gas-flow system
Nishihara; H.; Igarashi; Y.; Kodama; T., 筆頭著者, A marine obligately chemolithoautotrophic aerobic hydrogen-oxidizing bacterium Hydrogenovibrio marinus strain MH-110 was cultivated in a 2-l jar fermentor with continuous supply of a mixed gas of H-2/O2 /CO2. Fast growth began rapidly without a lag period, even in an atmosphere of 40% oxygen. Apparent specific growth rate of the initial stage of growth was 0.60-0.67 h-1 irrespective of any oxygen tension tested. Under an atmosphere containing hydrogen, the growth was promoted by the addition of tetrathionate to the medium. A dense culture was achieved by increasing the tension of oxygen and adding ferrous and magnesium ions before these factors became growth limiting. Under this culture condition, exponential growth continued until the cell density reached 14 g/l dry weight. The final cell concentration reached approximately 26 g/l dry weight after 30-h cultivation., SOC FERMENTATION BIOENGINEERING, JAPAN
Journal of Fermentation and Bioengineering, 1991年, ［査読有り］

A new isolate of Hydrogenobacter, an obligately chemolithoautotrophic, thermophilic, halophilic and aerobic hydrogen-oxidizing bacterium from seaside saline hot spring
Nishihara; H.; Igarashi; Y.; Kodama; T., 筆頭著者, SPRINGER VERLAG
Archives of Microbiology, 1990年, ［査読有り］

Isolation of an obligately chemolithoautotrophic, halophilic and aerobic hydrogen-oxidizing bacterium from marine environment
Nishihara; H.; Igarashi; Y.; Kodama; T., 筆頭著者, SPRINGER VERLAG
Archives of Microbiology, 1989年, ［査読有り］

Cell wall mutants of Saccharomyces cerevisiae with increased digestibility by cell wall lytic enzymes and protein extractability
Nakajima; T.; Konno; R.; Nishihara; H.; Matsuda; K., ELSEVIER SCIENCE INC
Journal of Fermentation Technology, 1988年, ［査読有り］

Structural alteration of cell wall polysaccharides from Saccharomyces cerevisiae
Nakajima; T.; Nishihara; H.; Chiba; Y.; Matsuda; K., ELSEVIER SCIENCE INC
Journal of Fermentation Technology, 1988年, ［査読有り］

好気性水素酸化細菌（水素細菌）とその利用
西原宏史
環境バイオテクノロジー学会誌, 2022年06月, ［査読有り］, ［招待有り］

低炭素社会をめざした好気的水素ガス代謝能の産業利用
西原宏史
バイオサイエンスとインダストリー, 2010年, ［査読有り］

好気性水素酸化細菌の生態および進化系統と利用の可能性
西原宏史
日本微生物生態学会誌, 2001年, ［査読有り］, ［招待有り］

光合成細菌・硫黄酸化細菌・水素酸化細菌の近縁な関係 -RuBiscoと16S rRNAの解析から-
西原宏史
日本生物工学会誌, 1996年, ［査読有り］, ［招待有り］

「独立栄養微生物によるCO2資源化技術」（監修 新井博之ら）、水素細菌によるグリコーゲンおよび養魚飼料用タンパク質資源の生産　　　　　　　　　　　　　　　
奥宏海、西原宏史, 共著
シーエムシー出版, 2023年12月13日
9784781317564

「微生物機能を活用した革新的生産技術の最前線 -ミニマムゲノムファクトリーとシステムバイオロジー-」（編集 清水昌ら）、ヒドロゲナーゼ反応のエネルギー供給系への応用　　　　　　　　　　　　　　　
西原宏史; 五十嵐泰夫; 石井正治, 共著
シーエムシー出版, 2007年12月14日
9784882319702

Genus III Hydrogenovibrio Nishihara, Igarashi and Kodama 1991b, 132VP　　　　　　　　　　　　　　　
Nishihara, H, 単著
Bergey's Manual of Systematic Bacteriology (2nd edn.), The Williams & Wilkins Co., Baltimore, 2005年

Biomass production from carbon dioxide by a marine hydrogen-oxidizing bacterium Hydrogenovibrio marinus　　　　　　　　　　　　　　　
Igarashi, Y; Nishihara, H; Kodama, T, 共著
Biochemical engineering for 2001(Proceedings of Asia-Pacific Biochemical Engineering Conference 1992), Springer-Verlag, Tokyo, 1992年

Biomass production from carbon dioxide by a marine hydrogen-oxidizing bacterium, Hydrogenovibrio marinus　　　　　　　　　　　　　　　
Nishihara, H; Igarashi, Y; Kodama, T, 共著
Microbial utilization of renewable resources(NRCT, NUS, DOST-JSPS Joint Seminor on Biotechnology), 1992年

土壌環境からの新たな水素酸化細菌の探索と分離
寺原直矢; 今泉尚志; 入江敬太; 長南茂; 西原宏史
日本農芸化学会2025年度大会, 2025年03月07日

水素酸化細菌の基礎と増殖
西原宏史
GIフォーラム第１回勉強会, 2024年11月14日

水素酸化細菌の生態と分離について　　　　　　　　　　　　　　　
西原宏史
日本農芸化学会2024年度大会 一般会員公募採択課題シンポジウム“CO2固定能を有する微生物とそのデータの利活用によるカーボンニュートラルへの貢献とバイオものづくり産業の活性化に向けて”, 2024年03月26日, ［招待有り］

海洋性水素酸化細菌Hydrogenovibrio marinus MH-110株の増殖におけるガス基質消費量の解析と新たな海洋性株の探索
野田 蒼士; 飯生 紗矢; 西原 宏史
第75回日本生物工学会大会, 2023年09月03日

海洋性水素酸化細菌Hydrogenovibrio marinus MH-110株の培養法の検討　　　　　　　　　　　　　　　
岡部航也; 松内愛美; 花田晶子; 春日友明; 明戸剛; 西原宏史
第75回日本生物工学会大会, 2023年09月03日

高い酸素耐性を示す水素酸化細菌Cupriavidus sp. FM-47株の分離とその特性解析　　　　　　　　　　　　　　　
第74回日本生物工学会大会, 2022年10月19日, 日本生物工学会
20221017, 20221020

強化されたCoA生合成による組換え大腸菌でのPHB生産　　　　　　　　　　　　　　　
工藤大嵩、朝山宗彦、西原宏史、長南茂
日本農芸化学会2022年度大会, 2022年03月17日, 日本農芸化学会
20220315, 20220318

水素細菌とその利用　　　　　　　　　　　　　　　
第73回日本生物工学会大会, 2021年10月28日, 日本生物工学会, ［招待有り］
20211027, 20211029

大腸菌のコエンザイムA増産株を用いたポリヒドロキシ酪酸生産　　　　　　　　　　　　　　　
小野渉,阿部健太,朝山宗彦,西原宏史,長南茂
日本微生物生態学会第33回大会, 2019年09月11日, 日本微生物生態学会

水素酸化細菌の培養特性と独立栄養代謝活性の解析　　　　　　　　　　　　　　　
近藤晃一郎、飯塚久美子、白神清三郎、石井正治、西原宏史
日本農芸化学会2016年度大会, 2016年03月30日, 日本農芸化学会

Hydrogenovibrio marinus由来膜結合型ヒドロゲナーゼのRalstonia eutrophaにおける発現　　　　　　　　　　　　　　　
遠藤遼平、川島真人、野口真治、梅村奈那実、西原宏史
第67回日本生物工学会大会, 2015年10月28日, 日本生物工学会

非白金水素エネルギー変換系につなげるヒドロゲナーゼの電気化学的アプローチ　　　　　　　　　　　　　　　
宋慶盛、河井翔太、北隅優希、白井理、西原宏史、加納健司
日本農芸化学会2014年度大会, 2014年03月29日, 日本農芸化学会

好気性水素細菌由来の酸素耐性型ヒドロゲナーゼ　　　　　　　　　　　　　　　
第8回茨城大学遺伝子実験施設公開シンポジウム“バイオ技術による次世代エネルギー研究最前線”, 2014年03月10日, 茨城大学遺伝子実験施設, ［招待有り］

水素細菌のヒドロゲナーゼ研究の新展開　　　　　　　　　　　　　　　
第65回日本生物工学会大会シンポジウム“無機化合物の微生物変換研究の最前線”, 2013年09月20日, 日本生物工学会, ［招待有り］

水素酸化細菌Hydrogenophilus thermoluteolusの培養特性と独立栄養代謝活性の解析　　　　　　　　　　　　　　　
飯塚久美子、石井正治、西原宏史
第65回日本生物工学会大会, 2013年09月18日, 日本生物工学

Carbon utilization in chemolithoautotrophs Hydrogenophilus thermoluteolus TH-1　　　　　　　　　　　　　　　
Nguyen Huu Tri; Hirofumi Nishihara; Hiroyuki Arai; Masaharu Ishii
第65回日本生物工学会大会, 2013年09月18日, 日本生物工学会

酸素耐性ヒドロゲナーゼの電気化学的活性化・不活性化反応の解析　　　　　　　　　　　　　　　
濱本塁、河井翔太、北隅優希、白井理、栗田耕平、西原宏史、加納健司
第58回ポーラログラフィーおよび電気分析化学討論会, 2012年11月27日, 日本ポーラログラフ学会（共催：日本化学会・電気化学会・日本分析化学会）

Hydrogenophilus属細菌の高温環境での広域な分布を支える酸化的リン酸化　　　　　　　　　　　　　　　
若井暁、西原宏史、三本木至宏
日本農芸化学会中四国支部第32回講演会, 2012年01月21日, 日本農芸化学会中四国支部

有機溶媒二層系における水素酸化細菌形質転換株による疎水性化合物の変換　　　　　　　　　　　　　　　
坂井可南子、小田航史、山本浩明、松山彰収、西原宏史
第63回日本生物工学会大会, 2011年09月26日, 日本生物工学会

水素利用微生物触媒の開発と水素による光学活性アルコール生産反応の解析　　　　　　　　　　　　　　　
宮田優、黒羽隼人、山本浩明、松山彰収、西原宏史
第63回日本生物工学会大会, 2011年09月26日, 日本生物工学会

Structural study of the O2-tolerant [NiFe] hydrogenase　　　　　　　　　　　　　　　
Y. Shomura; K. S. Yoon; H. Nishihara; Y. Higuchi
第49回日本生物物理学会年会, 2011年, 日本生物物理学会

Bioconversion of hydrophobic substrate by a transformant of H2-oxidizing bacterium using aqueous-organic solvent biphasic system　　　　　　　　　　　　　　　
Sakai, K; Nishihara, H
International Symposium on Asian Consortium for Sustainable Agriculture, 2010年12月

Hydrogenovibrio marinus由来膜結合型ヒドロゲナーゼ-シトクロム b複合体の精製と解析　　　　　　　　　　　　　　　
西原宏史、篠崎文治、会沢勇志、尹基石
第62回日本生物工学会大会, 2010年10月29日, 日本生物工学会

Hydrogenovibrio marinus由来膜結合型ヒドロゲナーゼによる電気化学反応の検討　　　　　　　　　　　　　　　
萩谷啓輔、佐々木唯、尹基石、西原宏史
第62回日本生物工学会大会, 2010年10月29日, 日本生物工学会

Analysis of H2-driven production of optically active alcohol using a transformant of hydrogen-oxidizing bacterium　　　　　　　　　　　　　　　
Miyata, Y; Nishihara, H
International Symposium on Asian Consortium for Sustainable Agriculture, 2010年10月

膜結合型[NiFe]ヒドロゲナーゼのX線結晶構造解析　　　　　　　　　　　　　　　
庄村康人、尹基石、西原宏史、樋口芳樹
平成22年度日本結晶学会年会, 2010年

好気性水素酸化細菌の水素ガス代謝能の応用　　　　　　　　　　　　　　　
西原宏史、尹基石
第61回日本生物工学会大会シンポジウム“独立栄養的代謝の産業応用的基軸”, 2009年09月25日, 日本生物工学会・バイオインダストリー協会新資源生物変換研究会共催, ［招待有り］

水素利用バイオプロセスによる環境調和型物質生産の検討　　　　　　　　　　　　　　　
西原宏史、五十嵐泰夫、石井正治
平成21年度日本農芸化学会大会シンポジウム“環境調和型微生物工場のデザインテクノロジー”, 2009年03月28日, 日本農芸化学会・バイオインダストリー協会新資源生物変換研究会共催, ［招待有り］

バイオプロセスへのヒドロゲナーゼの応用　　　　　　　　　　　　　　　
石井正治、尹基石、五十嵐泰夫、西原宏史
第60回日本生物工学会大会シンポジウム“新産業創出に挑むキーエンザイムの顔ぶれ”, 2008年08月27日, 日本生物工学会, ［招待有り］

Ralstonia eutrophaを宿主とする水素利用微生物触媒の開発と反応　　　　　　　　　　　　　　　
小田航史、松山彰収、山本浩明、西原宏史
第60回日本生物工学会大会, 2008年08月, 日本生物工学会

水素酸化細菌Rhodococcus opacusでのアルコール脱水素酵素の発現による水素利用微生物触媒の開発　　　　　　　　　　　　　　　
服部佑、松山彰収、山本浩明、橋本義輝、小林達彦、西原宏史
第60回日本生物工学会大会, 2008年08月, 日本生物工学会

海洋性水素酸化細菌由来膜結合型ヒドロゲナーゼのシトクロムb複合体の精製　　　　　　　　　　　　　　　
篠崎文治、会沢勇志、尹基石、西原宏史
2008年度日本蛋白質科学会, 2008年06月

好熱性水素細菌によるチオ硫酸酸化機構　　　　　　　　　　　　　　　
佐野涼子、西原宏史、亀谷将史、新井博之、石井正治、五十嵐泰夫、三本木至宏
日本農芸化学会中四国支部第20回講演会, 2008年01月

好気性水素酸化細菌由来ヒドロゲナーゼの構造安定性の解析　　　　　　　　　　　　　　　
家村恵美、尹基石、浜田寛之、白木賢太郎、西原宏史
2007年度日本蛋白質科学会, 2007年05月

Hydrogenovibrio marinusおよびRalstonia eutropha由来ヒドロゲナーゼの精製とCDスペクトル測定を用いた構造安定性の解析　　　　　　　　　　　　　　　
家村恵美、尹基石、浜田寛之、白木賢太郎、西原宏史
日本農芸化学会2007年度大会, 2007年03月, 日本農芸化学会

Application of the hydrogen-activating enzyme, hydrogenase, for development of biological hydrogen technologies and an aspect of hydrogenase stabilization　　　　　　　　　　　　　　　
Nishihara, H
International Symposium on Sustainable Agriculture in Asia -Challenges for Agricultural Sciences on Environmental Problems under Global Changes-, 2006年09月, Ibaraki Univ., Bogor Univ.

Thiosulfate Oxidation by a Moderate Thermophilic Hydrogen-Oxidizing Bacterium, Hydrogenophilus thermoluteolus　　　　　　　　　　　　　　　
Miyake, D; Ichiki, S; Tanabe, M; Oda, T; Kuroda, H; Nishihara, H; Sambongi, Y
11th International Symposium on Microbial Ecology, 2006年08月

NAD還元型ヒドロゲナーゼとアルコール脱水素酵素との共役による水素駆動型物質変換反応の検討　　　　　　　　　　　　　　　
小田高広、山本浩明、松山彰収、石井正治、五十嵐泰夫、西原宏史
日本農芸化学会2006年度大会, 2006年03月, 日本農芸化学会

高温性水素細菌Hydrogenophilus thermoluteolusのATP合成酵素の活性測定と遺伝子解析　　　　　　　　　　　　　　　
池田拓未、山口奈穂、西原宏史、三本木至宏
日本農芸化学会中四国支部第16回講演会, 2006年, 日本農芸化学会

高温性水素細菌Hydrogenophilus thermoluteolusの硫黄代謝機構　　　　　　　　　　　　　　　
三本木至宏、田辺京、西原宏史
日本農芸化学会中四国支部第16回講演会, 2006年, 日本農芸化学会

高温性水素細菌Hydrogenophilus thermoluteolusのATP合成酵素の活性測定と遺伝子解析　　　　　　　　　　　　　　　
池田拓未、山口奈穂、西原宏史、三本木至宏
日本農芸化学会2006年度大会, 2006年, 日本農芸化学会

高温性水素細菌Hydrogenophilus thermoluteolusの硫黄酸化酵素群の解析　　　　　　　　　　　　　　　
三宅大輔、小田高広、西原宏史、三本木至宏
日本農芸化学会中四国支部第14回講演会, 2006年, 日本農芸化学会

Hydrogenophilus thermoluteolus TH-1由来のNAD還元型ヒドロゲナーゼの生化学的解析　　　　　　　　　　　　　　　
尹基石、酒井由紀子、船木強、藤澤 清史、石井正治、五十嵐泰夫、西原宏史
日本生物工学会大会, 2005年, 日本生物工学会

好熱性水素細菌Hydrogenobacter thermophilus TK-6株由来の膜結合型ヒドロゲナーゼの特性解析　　　　　　　　　　　　　　　
浦崎尚、尹基石、西原宏史、松本伯夫、大村直也、新井博之、石井正治、五十嵐泰夫
日本農芸化学会大会, 2005年, 日本農芸化学会

好熱菌Hydrogenophilus thermoluteolusの硫黄酸化酵素の機能解析　　　　　　　　　　　　　　　
三宅大輔、小田高広、黒田久雄、西原宏史、三本木至宏
日本農芸化学会大会, 2005年, 日本農芸化学会

シアノバクテリア及び藻類の光水素発生能の改善を目指した自己集積型ヒドロゲナーゼ-PSI複合体の設計　　　　　　　　　　　　　　　
伊原正喜、西原宏史、仲本 準、小島 幸治、本間大奨、蒲池利章、大倉 一郎、前田瑞夫
日本農芸化学会大会, 2005年, 日本農芸化学会

Hydrogenophilus thermoluteolus TH-1由来のNAD還元型ヒドロゲナーゼの性質と遺伝子解析　　　　　　　　　　　　　　　
酒井由紀子、尹基石、舩木強、藤澤清史、石井正治、五十嵐泰夫、西原宏史
日本農芸化学会大会, 2005年, 日本農芸化学会

Phylogenetic diversity of bacterial community in the Mt. Pinatubo pyroclastic deposits　　　　　　　　　　　　　　　
Sato, Y; Ogiwara, K; Nishihara, H; Yoshida, M; Watanabe, M; Ohta, H
10th International Symposium on Microbial Ecology, 2004年08月

Analysis of the thermostable membrane-bound hydrogenase from Hydrogenovibrio marinus and an attempt for hydrogenase stabilization　　　　　　　　　　　　　　　
Nishihara, H; Yoon, K. S; Ogata, H; Higuchi, Y; DeLacey, A. L; Fernández, V. M; Bernhard, M; Pohlmann, A; Friedrich, B
15th World Hydrogen Energy Conference, 2004年06月, ［招待有り］

シアノバクテリア及び藻類の光水素発生能の改善を目指した自己集積型ヒドロゲナーゼ-PSI複合体の設計　　　　　　　　　　　　　　　
伊原正喜、西原宏史、仲本準、小島幸治、本間大輔、蒲池利章、大倉一郎
第27回日本分子生物学会年会, 2004年, 日本分子生物学会

好熱性水素細菌Hydrogenobacter thermophilus TK-6株由来のpyruvate:ferredoxin oxidoreductaseによる炭酸固定反応　　　　　　　　　　　　　　　
池田丈、山本正浩、尹基石、西原宏史、新井博之、石井正治、五十嵐泰夫
日本農芸化学会大会, 2004年, 日本農芸化学会

新規に分離されたHydrogenophaga sp. AH-24の膜結合型ヒドロゲナーゼの精製と性質　　　　　　　　　　　　　　　
尹基石、塚田なつき、藤澤清史、石井正治、五十嵐泰夫、西原宏史
日本生物工学会大会, 2003年, 日本生物工学会

Hydrogenophilus thermoluteolus TH-1株由来のNAD還元型ヒドロゲナーゼの精製と性質　　　　　　　　　　　　　　　
舩木強、尹基石、藤澤清史、石井正治、五十嵐泰夫、西原宏史
日本生物工学会大会, 2003年, 日本生物工学会

Ralstonia eutropha由来膜結合型ヒドロゲナーゼへのIle残基導入による安定化効果の解析　　　　　　　　　　　　　　　
福田景一・寺下麻美、井上美帆、緒方英明、樋口芳樹、尹基石、西原宏史
日本生物工学会大会, 2003年, 日本生物工学会

海洋性水素酸化細菌の膜結合型ヒドロゲナーゼ関連遺伝子群の解析　　　　　　　　　　　　　　　
川又寛和、M. Bernhard、A. Pohlmann、B. Friedrich、尹基石、西原宏史
日本農芸化学会大会, 2003年, 日本農芸化学会

Occurrence of hydrogen-oxidizing spp. As a primary microbe in the Pinatubo pyroclastic deposite　　　　　　　　　　　　　　　
Sato, Y; Ogiwara, K; Nishihara, H; Yoshida, M; Watanabe, M; Ohta, H
16th International Symposium on Environmental Biogeochemistry, 2003年

イソロイシン残基の導入によるヒドロゲナーゼの耐熱化　　　　　　　　　　　　　　　
西原宏史他
日本農芸化学会大会, 2002年, 日本農芸化学会

水素の生物循環・代謝の新視点　　　　　　　　　　　　　　　
西原宏史
学際シンポジウム 物質循環と代謝の新視点, 2002年, 日本微生物生態学会・日本生物工学会共催, ［招待有り］

フィリピン・ピナツボ火山泥流堆積物から分離した細菌の栄養代謝特性　　　　　　　　　　　　　　　
佐藤嘉則、西原宏史、吉田正夫、渡辺真紀子、太田寛行
日本微生物生態学会大会, 2001年, 日本微生物生態学会

海洋性水素酸化細菌の耐熱・耐酸素性ヒドロゲナーゼ遺伝子のシークエンスと蛋白質一次配列における特徴　　　　　　　　　　　　　　　
西原宏史、M. Bernhard、A. Pohlmann、B. Friedrich、高村義親
日本農芸化学会大会, 2001年, 日本農芸化学会

Sequence of an extremely thermophilic membrane-bound hydrogenase (MBH) from Hydrogenovibrio merinus and stabilization of Ralstonia eutropha MBH by amino acid substitutions　　　　　　　　　　　　　　　
Nishihara, H; Bernhard, M; Pohlmann, A; Igarashi, Y; Takamura, Y; Friedrich, B
6th International Conference on the Molecular Biology of Hydrogenases, 2000年

Sequences and organization of genes of an extremely thermophilic and oxygen-stable membrane-bound hydrogenase from Hydrogenovibrio marinus　　　　　　　　　　　　　　　
Nishihara, H; Bernhard, M; Pohlmann, A; Takamura, Y; Friedrich, B
Microbioiogy 2000 -Health, Ecology, Biotechnology-, 2000年

水素酸化細菌の生態および進化系統と利用の可能性　　　　　　　　　　　　　　　
西原宏史
日本微生物生態学会大会シンポジウム, 2000年, 日本微生物生態学会, ［招待有り］

Pseudomonas putidaのマロン酸脱炭酸酵素遺伝子のクローニング　　　　　　　　　　　　　　　
長南茂、久留主泰朗、西原宏史、高村義親
日本農芸化学会大会, 1999年, 日本農芸化学会

ヒドロゲナーゼ研究の最近の動向　　　　　　　　　　　　　　　
西原宏史
未来へのバイオ技術勉強会, 1999年, (財)バイオインダストリー協会, ［招待有り］

Pseudomonas putidaのマロン酸脱炭酸酵素の反応機構　　　　　　　　　　　　　　　
長南茂、高木俊和、西原宏史、高村義親
日本農芸化学会大会, 1998年, 日本農芸化学会

海洋性水素酸化細菌のヒドロゲナーゼ分子種の解析　　　　　　　　　　　　　　　
西原宏史、宮田佳典、宮下洋士、青山勝博、五十嵐泰夫、高村義親
日本農芸化学会大会, 1998年

好熱性光合成細菌の探索とその水素生産への応用　　　　　　　　　　　　　　　
西原宏史、久永信吾、岩崎哲朗、高村義親
日本農芸化学会大会, 1998年, 日本農芸化学会

Microcystis属とOscillatoria属シアノバクテリアの各種有機物に対する増殖応答の相違　　　　　　　　　　　　　　　
千野まり子、長田綾子、岡本晃司、西原宏史、矢木修身、高村義親
日本農芸化学会大会, 1998年, 日本農芸化学会

ウロン酸資化菌によるポリ-β-ヒドロキシ酪酸(PHB)および菌体外多糖の生産　　　　　　　　　　　　　　　
西原宏史、深見健、小林幹彦、高村義親
日本生物工学会大会, 1997年, 日本生物工学会

An extremely thermophilic and oxygen-stable membrane-bound hydrogenase from a marine hydrogen-oxidizing bacterium Hydrogenovibrio marinus　　　　　　　　　　　　　　　
Nishihara, H; Miyashita, Y; Aoyama, K; Miyata, Y; Kodama, T; Igarashi, Y; Takamura, Y
5th International Conference on the Molecular Biology of Hydrogenases, 1997年

海洋性水素酸化細菌の好熱性ヒドロゲナーゼの性質　　　　　　　　　　　　　　　
西原宏史、宮下洋士、青山勝博、五十嵐泰夫、児玉徹、高村義親
日本農芸化学会大会, 1996年, 日本農芸化学会

Research strategies for biological hydrogen gas production by nitrogenase and hydrogenase -from ammonia to hydrogen gas-　　　　　　　　　　　　　　　
Watanabe, I; Asada, Y; Kumazawa, K; Miyamoto, H; Nishihara, H; Ohmiya K; Onodera, K; Uozumi, T
7th International Symposium on Nitrogen Fixation with Non-legumes, 1996年

Microcystis属シアノバクテリアのRAPD法によるDNA多型解析　　　　　　　　　　　　　　　
西原宏史、三輪洋康、高村義親、長島寛、渡辺真之、矢木修身
日本農芸化学会大会, 1995年, 日本農芸化学会

グルクロン酸資化菌の分離・同定とバイオマス変換への利用　　　　　　　　　　　　　　　
西原宏史、清水順也、平田愛子、小林幹彦、高村義親
日本農芸化学会大会, 1995年, 日本農芸化学会

好（耐）アルコール性乳酸菌L. homohiochiiの各種アルコールに対する増殖特性と低分子物質の細胞外蓄積について　　　　　　　　　　　　　　　
安田正博、西原宏史、高村義親
日本農芸化学会大会, 1995年

Pseudomonas ovalisのマロン酸脱炭酸酵素の構造と機能　　　　　　　　　　　　　　　
藤尾徹、高村義親、許晴台、西原宏史
日本農芸化学会大会, 1995年, 日本農芸化学会

霞ヶ浦における藻類種の優占機構　　　　　　　　　　　　　　　
矢木修身、岩崎一弘、内山裕夫、高村義親、西原宏史
日本農芸化学会大会, 1995年

海洋性独立栄養細菌H. marinus MH-110の持つ複数のRuBisCOについて　　　　　　　　　　　　　　　
S. Y. Chun、矢口敏昭、西原宏史、五十嵐泰夫、児玉徹
日本農芸化学会関東支部大会, 1992年, 日本農芸化学会

Hydrogenovibrio marinus MH-110株のRuBisCO遺伝子の塩基配列　　　　　　　　　　　　　　　
S. Y. Chun、西原宏史、五十嵐泰夫、児玉徹
日本農芸化学会大会, 1992年, 日本農芸化学会

RuBisCOs from an obligately chemolithoautotrophic, marine hydrogen-oxidizing bacterium, Hydrogenovibrio marinus　　　　　　　　　　　　　　　
Igarashi, Y; Chung, S; Nishihara, H; Kodama, T
International Symposium on Microbial growth on C1 compounds, 1992年

Biomass production from carbon dioxide by a marine hydrogen-oxidizing bacterium Hydrogenovibrio marinus　　　　　　　　　　　　　　　
Igarashi, Y; Nishihara, H; Kodama, T
Asia-Pacific Biochemical Engineering Conference '92, 1992年

Biomass production from carbon dioxide by a marine ,hydrogen-oxidizing bacterium Hydrogenovibrio marinus　　　　　　　　　　　　　　　
Igarashi; Y.; Nishihara; H.; Kodama; T.
Asia-Pacific Biochemical Engineering Conference '92, 1992年

Hydrogenovibrio marinus MH-110株からのL2型RuBisCOの精製及び性質　　　　　　　　　　　　　　　
S. Y. Chun、西原宏史、五十嵐泰夫、児玉徹
日本発酵工学会大会, 1991年, 日本発酵工学会

海洋性水素細菌Hydrogenovibrio marinus gen. nov., sp. nov.による炭酸ガスからのグルコースポリマーの生産　　　　　　　　　　　　　　　
西原宏史、五十嵐泰夫、児玉徹、中島佑
日本発酵工学会大会, 1991年, 日本発酵工学会

海洋性オートトローフMH-110株のRuBisCO遺伝子のクローニングと発現　　　　　　　　　　　　　　　
S. Y. Chun、西原宏史、五十嵐泰夫、児玉徹
日本発酵工学会大会, 1990年, 日本発酵工学会

絶対独立栄養性を示す好塩性水素細菌2菌株の分類　　　　　　　　　　　　　　　
西原宏史、五十嵐泰夫、児玉徹
日本農芸化学会大会, 1990年, 日本農芸化学会

海洋性水素細菌の培養特性と高濃度培養　　　　　　　　　　　　　　　
西原宏史、五十嵐泰夫、児玉徹
日本発酵工学会大会, 1989年, 日本発酵工学会

Isolation of obligately autotrophic hydrogen bacteria from saline environments　　　　　　　　　　　　　　　
Nishihara, H; Igarashi, Y; Kodama, T
6th International Symposium on Microbial growth on C1 compounds, 1989年

好熱性好塩性水素細菌の単離と性質　　　　　　　　　　　　　　　
西原宏史、五十嵐泰夫、児玉徹
日本農芸化学会大会, 1988年, 日本農芸化学会

Obligate autotrophic hydrogen bacteria from halophilic environment　　　　　　　　　　　　　　　
Kodama, T; Igarashi, Y; Nishihara, H
8th International Biotechnology Symposium, 1988年

海洋性水素細菌の単離と性質　　　　　　　　　　　　　　　
西原宏史、五十嵐泰夫、児玉徹
日本発酵工学会大会, 1987年, 日本発酵工学会

Saccharomyces cerevisiae細胞壁変異株の化学的性質(III)　　　　　　　　　　　　　　　
中島佑、西原宏史、松田和雄
日本農芸化学会大会, 1987年, 日本農芸化学会

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