Updated on 2024/11/10

写真a

 
AZAI Chihiro
 
Organization
Faculty of Science and Engineering Associate Professor
Other responsible organization
Biological Sciences Course of Graduate School of Science and Engineering, Master's Program
Biological Sciences Course of Graduate School of Science and Engineering, Doctoral Program
Contact information
The inquiry by e-mail is 《here
External link

Degree

  • 博士(理学) ( 大阪大学 )

  • 修士(理学) ( 大阪大学 )

Education

  • 2010.6
     

    Osaka University   doctor course   completed

  • 2007.3
     

    Osaka University   master course   completed

  • 2005.3
     

    Kobe University   graduated

Research History

  • 2023.4 - Now

    Chuo University   Department of Biological Sciences, Faculty of Science and Engineering   Associate Professor

  • 2018.4 - 2023.3

    Ritsumeikan University   Department of Bioinformatics, College of Llife Sciences   Lecturer (Assistant Professor)

  • 2013.4 - 2018.3

    Ritsumeikan University   Department of Bioinformatics, College of Llife Sciences   Assistant Professor

  • 2011.4 - 2013.3

    Nagoya University   Graduate School of Science   Research Fellow (PD) of the Japan Society for the Promotion of Science

  • 2009.4 - 2011.3

    Osaka University   Graduate School of Science   Research Fellow (DC) of the Japan Society for the Promotion of Science

Professional Memberships

  • International Society of Photosynthesis Research

  • The Japanese Society of Plant Physiologists

  • The Japanese Society of Photosynthesis Research

  • The Biophysical Society of Japan

Research Interests

  • Photosynthesis, Electron transfer, Molecular evolution, Photosynthetic bacteria

Research Areas

  • Life Science / Evolutionary biology

  • Life Science / Plant molecular biology and physiology

  • Life Science / Applied microbiology

  • Life Science / Applied biochemistry

  • Life Science / Functional biochemistry

  • Life Science / Genome biology

  • Life Science / Structural biochemistry

  • Life Science / Molecular biology

  • Life Science / Biophysics

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Papers

  • Genes for the type-I reaction center and galactolipid synthesis are required for chlorophyll a accumulation in a purple photosynthetic bacterium. Invited Reviewed International journal

    Yusuke Tsukatani, Chihiro Azai, Tomoyasu Noji, Shigeru Kawai, Saori Sugimoto, Shigeru Shimamura, Yasuhiro Shimane, Jiro Harada, Tadashi Mizoguchi, Hitoshi Tamiaki, Shinji Masuda

    Plant & cell physiology   2024.7

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

    Anoxygenic photosynthesis is diversified into two classes: chlorophototrophy based on a bacterial type-I or type-II reaction center (RC). Whereas the type-I RC contains both bacteriochlorophyll and chlorophyll, type-II RC-based phototrophy relies only on bacteriochlorophyll. However, type-II phototrophic bacteria theoretically have the potential to produce chlorophyll a by the addition of an enzyme, chlorophyll synthase, because the direct precursor for the enzyme, chlorophyllide a, is produced as an intermediate of BChl a biosynthesis. In this study, we attempted to modify the type-II proteobacterial phototroph Rhodovulum sulfidophilum to produce chlorophyll a by introducing chlorophyll synthase, which catalyzes the esterification of a diterpenoid group to chlorophyllide a thereby producing chlorophyll a. However, the resulting strain did not accumulate chlorophyll a, perhaps due to absence of endogenous chlorophyll a-binding proteins. We further heterologously incorporated genes encoding the type-I RC complex to provide a target for chlorophyll a. Heterologous expression of type-I RC subunits, chlorophyll synthase, and galactolipid synthase successfully afforded detectable accumulation of chlorophyll a in Rdv. sulfidophilum. This suggests that the type-I RC can work to accumulate chlorophyll a and that galactolipids are likely necessary for the type-I RC assembly. The evolutionary acquisition of type-I RCs could be related to prior or concomitant acquisition of galactolipids and chlorophylls.

    DOI: 10.1093/pcp/pcae076

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  • Illuminating the coevolution of photosynthesis and Bacteria Reviewed International journal

    Arisa Nishihara, Yusuke Tsukatani, Chihiro Azai, Masaru K. Nobu

    Proceedings of the National Academy of Sciences   121 ( 25 )   2024.6

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Proceedings of the National Academy of Sciences  

    Life harnessing light energy transformed the relationship between biology and Earth—bringing a massive flux of organic carbon and oxidants to Earth’s surface that gave way to today’s organotrophy- and respiration-dominated biosphere. However, our understanding of how life drove this transition has largely relied on the geological record; much remains unresolved due to the complexity and paucity of the genetic record tied to photosynthesis. Here, through holistic phylogenetic comparison of the bacterial domain and all photosynthetic machinery (totally spanning >10,000 genomes), we identify evolutionary congruence between three independent biological systems—bacteria, (bacterio)chlorophyll-mediated light metabolism (chlorophototrophy), and carbon fixation—and uncover their intertwined history. Our analyses uniformly mapped progenitors of extant light-metabolizing machinery (reaction centers, [bacterio]chlorophyll synthases, and magnesium-chelatases) and enzymes facilitating the Calvin–Benson–Bassham cycle (form I RuBisCO and phosphoribulokinase) to the same ancient Terrabacteria organism near the base of the bacterial domain. These phylogenies consistently showed that extant phototrophs ultimately derived light metabolism from this bacterium, the last phototroph common ancestor (LPCA). LPCA was a non-oxygen-generating (anoxygenic) phototroph that already possessed carbon fixation and two reaction centers, a type I analogous to extant forms and a primitive type II. Analyses also indicate chlorophototrophy originated before LPCA. We further reconstructed evolution of chlorophototrophs/chlorophototrophy post-LPCA, including vertical inheritance in Terrabacteria, the rise of oxygen-generating chlorophototrophy in one descendant branch near the Great Oxidation Event, and subsequent emergence of Cyanobacteria. These collectively unveil a detailed view of the coevolution of light metabolism and Bacteria having clear congruence with the geological record.

    DOI: 10.1073/pnas.2322120121

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  • Experimental and Theoretical Mutation of Exciton States on the Smallest Type-I Photosynthetic Reaction Center Complex of a Green Sulfur Bacterium Chlorobaclum tepidum Reviewed International journal

    Akihiro Kimura, Hirotaka Kitoh-Nishioka, Hirozo Oh-oka, Shigeru Itoh, Chihiro Azai

    The Journal of Physical Chemistry B   2024.1

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    DOI: 10.1021/acs.jpcb.3c07424

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  • 緑色硫黄細菌の光合成による水素およびメタンの生成 Invited Reviewed

    浅井智広

    触媒(Catalysts and Catalysis)   65 ( 5 )   282 - 286   2023.10

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    Authorship:Lead author, Corresponding author   Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:一般社団法人 触媒学会  

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  • Soluble domains of cytochrome c-556 and Rieske iron–sulfur protein from Chlorobaculum tepidum: Crystal structures and interaction analysis Reviewed International journal

    Hiraku Kishimoto, Chihiro Azai, Tomoya Yamamoto, Risa Mutoh, Tetsuko Nakaniwa, Hideaki Tanaka, Yohei Miyanoiri, Genji Kurisu, Hirozo Oh-oka

    Current Research in Structural Biology   5   100101 - 100101   2023.4

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

    DOI: 10.1016/j.crstbi.2023.100101

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  • Photosynthesis | Green Bacteria – Energy Transfer and Electron Transport Invited Reviewed

    Hirozo Oh-oka, Jiro Harada, Chihiro Azai

    Encyclopedia of Biological Chemistry III   2   333 - 351   2021

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    Language:English   Publishing type:Part of collection (book)   Publisher:Elsevier  

    DOI: 10.1016/b978-0-12-819460-7.00031-1

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  • Anaerobic energy dissipation by glycosylated carotenoids in the green sulfur bacterium Chlorobaculum tepidum Invited Reviewed International journal

    Chihiro Azai, Jiro Harada, Shogo Fujimoto, Shinji Masuda, Daisuke Kosumi

    Journal of Photochemistry and Photobiology A: Chemistry   403   112828 - 112828   2020.12

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    Authorship:Lead author, Corresponding author   Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    DOI: 10.1016/j.jphotochem.2020.112828

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  • Energy transfer and primary charge separation upon selective femtosecond excitation at 810 nm in the reaction center complex from Heliobacterium modesticaldum Reviewed

    Risa Kojima, Hayata Yamamoto, Chihiro Azai, Chiasa Uragami, Hideki Hashimoto, Daisuke Kosumi, Hirozo Oh-oka

    Journal of Photochemistry and Photobiology A: Chemistry   401   112758   2020.10

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

    DOI: 10.1016/j.jphotochem.2020.112758

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  • Mutation of alanine-422 in KaiC leads to a low amplitude of rhythm in the reconstituted cyanobacterial circadian clock. Reviewed

    Kazuki Nagata, Katsuaki Oyama, Atsushi Ota, Chihiro Azai, Kazuki Terauchi

    The Journal of general and applied microbiology   66 ( 2 )   140 - 146   2020.6

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    The cyanobacterial circadian oscillator can be reconstituted by mixing the purified clock proteins KaiA, KaiB, and KaiC with ATP in vitro, leading to a 24-h oscillation of KaiC phosphorylation. The cyanobacterial mutant pr1 carrying valine instead of alanine at position 422 of KaiC (KaiC-A422V) lost the ability to shift the phase of the circadian rhythm. In this study, we analyzed KaiC-A422V to investigate the effect of this single-residue substitution on the in vitro reconstitution of KaiC oscillation. KaiC-A422V exhibited low amplitude oscillations of phosphorylation with a smaller amount of Kai complex than wild-type KaiC (KaiC-WT). Although KaiA can stimulate KaiC phosphorylation, the phosphorylation level of KaiC-A422V is much lower than that of KaiC-WT even at higher KaiA concentrations. It has been suggested that monomer shuffling of KaiC is involved in entraining the in vitro rhythm. To examine whether KaiC-A422V has the capacity for monomer shuffling, we used the difference in the amplitude of the phosphorylation rhythms between KaiC-WT and KaiC-A422V as the indicator of monomer shuffling. When KaiC-A422V and KaiC-WT were mixed, the amplitude of the phosphorylation rhythm changed according to the mixing ratio. This suggests that KaiC-A422V has a reduced ability to shuffle monomers in hexameric KaiC. In addition, the A422V mutation resulted in a change of the stability of the KaiC protein.

    DOI: 10.2323/jgam.2020.01.008

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  • Synechocystis KaiC3 Displays Temperature- and KaiB-Dependent ATPase Activity and Is Important for Growth in Darkness Reviewed International coauthorship

    Anika Wiegard, Christin Köbler, Katsuaki Oyama, Anja K. Dörrich, Chihiro Azai, Kazuki Terauchi, Annegret Wilde, Ilka M. Axmann

    Journal of Bacteriology   202 ( 4 )   e00478-19   2020.1

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

    The circadian clock influences the cyanobacterial metabolism, and deeper understanding of its regulation will be important for metabolic optimizations in the context of industrial applications. Due to the heterogeneity of cyanobacteria, characterization of clock systems in organisms apart from the circadian model Synechococcus elongatus PCC 7942 is required. Synechocystis sp. strain PCC 6803 represents a major cyanobacterial model organism and harbors phylogenetically diverged homologs of the clock proteins, which are present in various other noncyanobacterial prokaryotes. By our in vitro studies we unravel the interplay of the multiple Synechocystis Kai proteins and characterize enzymatic activities of the nonstandard clock homolog KaiC3. We show that the deletion of kaiC3 affects growth in constant darkness, suggesting its involvement in the regulation of nonphotosynthetic metabolic pathways.

    DOI: 10.1128/jb.00478-19

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  • Heliobacterium modesticaldum光合成反応中心への選択励起における励起エネルギー移動および電子移動反応 Invited

    小島理沙, 山元颯太, 浅井智広, 小澄大輔, 大岡宏造

    2019.12

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  • X-ray structure of the type-I reaction center from Heliobacterium modesticaldum at 3.2 A resolution Invited

    NAKANIWA Tetsuko, MUTOH Risa, FUSHIMI Kokoro, YASUDA Aya, MIZOGUCHI Tadashi, TAMIAKI Hitoshi, AZAI Chihiro, TANAKA Hideaki, ITOH Shigeru, OH-OKA Hirozo, KURISU Genji

    生物物理(Web)   58 ( Supplement 1-2 )   2018.9

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    Language:English   Publishing type:Research paper (other academic)  

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  • Soft X-Ray Imaging of Cellular Carbon and Nitrogen Distributions in Heterocystous Cyanobacteria Reviewed

    Takahiro Teramoto, Chihiro Azai, Kazuki Terauchi, Masashi Yoshimura, Toshiaki Ohta

    Plant Physiology   177 ( 1 )   52 - 61   2018.3

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

    DOI: 10.1104/pp.17.01767

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  • Light-Induced Electron Spin-Polarized (ESP) EPR Signal of the P800+ Menaquinone Radical Pair State in Oriented Membranes of Heliobacterium modesticaldum: Role/Location of Menaquinone in the Homodimeric Type I Reaction Center Reviewed

    Toru Kondo, Masahiro Matsuoka, Chihiro Azai, Masami Kobayashi, Shigeru Itoh, Hirozo Oh-oka

    The Journal of Physical Chemistry B   122 ( 9 )   2536 - 2543   2018.2

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:American Chemical Society (ACS)  

    DOI: 10.1021/acs.jpcb.7b12171

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  • Rapid C8-vinyl reduction of divinyl-chlorophyllide a by BciA from Rhodobacter capsulatus Reviewed

    Chihiro Azai, Manami Kobayashi, Tadashi Mizoguchi, Hitoshi Tamiaki, Kazuki Terauchi, Yusuke Tsukatani

    Journal of Photochemistry and Photobiology A: Chemistry   353   661 - 666   2018.2

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

    DOI: 10.1016/j.jphotochem.2017.09.010

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  • 軟X線顕微鏡による生きたシアノバクテリア中の細胞内元素分布の可視化

    寺本高啓, 浅井智広, 吉村真史, 寺内一姫, 太田俊明

    日本放射光学会年会・放射光科学合同シンポジウム(Web)   31st   2018

  • Phosphorylation at Thr432 induces structural destabilization of the CII ring in the circadian oscillator KaiC Reviewed

    Katsuaki Oyama, Chihiro Azai, Jun Matsuyama, Kazuki Terauchi

    FEBS Letters   592 ( 1 )   36 - 45   2017.12

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

    DOI: 10.1002/1873-3468.12945

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    Other Link: https://onlinelibrary.wiley.com/doi/full-xml/10.1002/1873-3468.12945

  • Determination of carbon-to-nitrogen ratio in the filamentous and heterocystous cyanobacterium Anabaena sp. PCC 7120 with single-cell soft X-ray imaging Reviewed

    T Teramoto, M Yoshimura, C Azai, K Terauchi, T Ohta

    Journal of Physics: Conference Series   849   012005 - 012005   2017.6

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    Language:English   Publishing type:Research paper (international conference proceedings)   Publisher:IOP Publishing  

    DOI: 10.1088/1742-6596/849/1/012005

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    Other Link: http://stacks.iop.org/1742-6596/849/i=1/a=012005?key=crossref.17476d78fccf1c31f9ed592a8f34b2fd

  • BN-PAGE analysis of cyanobacterial clock protein KaiC Invited Reviewed

    Kazuki Terauchi, Katsuaki Oyama, Chihiro Azai

    Electrophoresis Letters   61 ( 2 )   107 - 110   2017

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    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:Japanese Electrophoresis Society  

    DOI: 10.2198/electroph.61.107

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  • Conversion between two conformational states of KaiC is induced by ATP hydrolysis as a trigger for cyanobacterial circadian oscillation Reviewed

    Katsuaki Oyama, Chihiro Azai, Kaori Nakamura, Syun Tanaka, Kazuki Terauchi

    Scientific Reports   6 ( 1 )   32443   2016.9

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

    Abstract

    The cyanobacterial circadian oscillator can be reconstituted in vitro by mixing three clock proteins, KaiA, KaiB and KaiC, with ATP. KaiC is the only protein with circadian rhythmic activities. In the present study, we tracked the complex formation of the three Kai proteins over time using blue native (BN) polyacrylamide gel electrophoresis (PAGE), in which proteins are charged with the anionic dye Coomassie brilliant blue (CBB). KaiC was separated as three bands: the KaiABC complex, KaiC hexamer and KaiC monomer. However, no KaiC monomer was observed using gel filtration chromatography and CBB-free native PAGE. These data indicate two conformational states of KaiC hexamer and show that the ground-state KaiC (gs-KaiC) is stable and competent-state KaiC (cs-KaiC) is labile and degraded into monomers by the binding of CBB. Repeated conversions from gs-KaiC to cs-KaiC were observed over 24 h using an in vitro reconstitution system. Phosphorylation of KaiC promoted the conversion from gs-KaiC to cs-KaiC. KaiA sustained the gs-KaiC state, and KaiB bound only cs-KaiC. An E77Q/E78Q-KaiC variant that lacked N-terminal ATPase activity remained in the gs-KaiC state. Taken together, ATP hydrolysis induces the formation of cs-KaiC and promotes the binding of KaiB, which is a trigger for circadian oscillations.

    DOI: 10.1038/srep32443

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    Other Link: https://www.nature.com/articles/srep32443

  • Orientations of Iron–Sulfur Clusters FA and FB in the Homodimeric Type-I Photosynthetic Reaction Center of Heliobacterium modesticaldum Reviewed

    Toru Kondo, Masahiro Matsuoka, Chihiro Azai, Shigeru Itoh, Hirozo Oh-oka

    The Journal of Physical Chemistry B   120 ( 18 )   4204 - 4212   2016.5

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    DOI: 10.1021/acs.jpcb.6b01112

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  • Nitrogen mapping of the filamentous and heterocystous cyanobacterium Anabaena sp. PCC 7120 cells by soft X-ray microscopy Invited

    T. Teramoto, M. Yoshimura, C. Azai, K. Terauchi, H. Namba, T. Ohta

    Memoirs of the SR center Ritsumeikan University   18   125 - 126   2016.5

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  • Rethinking about the evolution toward oxygenic photosynthesis and experiments to demonstrate it in the laboratory Invited Reviewed

    C. Azai, Y. Tsukatani

    News letter of the Japanese Society of Photosynthesis Research   26 ( 1 )   43 - 58   2016.4

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    Authorship:Lead author, Corresponding author   Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:The Japanese Society of Photosynthesis Research  

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  • Mutation-induced perturbation of the special pair P840 in the homodimeric reaction center in green sulfur bacteria Reviewed

    Chihiro Azai, Yuko Sano, Yuki Kato, Takumi Noguchi, Hirozo Oh-oka

    Scientific Reports   6 ( 1 )   19878   2016.1

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    Authorship:Lead author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Springer Science and Business Media LLC  

    Abstract

    Homodimeric photosynthetic reaction centers (RCs) in green sulfur bacteria and heliobacteria are functional homologs of Photosystem (PS) I in oxygenic phototrophs. They show unique features in their electron transfer reactions; however, detailed structural information has not been available so far. We mutated PscA-Leu688 and PscA-Val689 to cysteine residues in the green sulfur bacterium Chlorobaculum tepidum; these residues were predicted to interact with the special pair P840, based on sequence comparison with PS I. Spectroelectrochemical measurements showed that the L688C and V689C mutations altered a near-infrared difference spectrum upon P840 oxidation, as well as the redox potential of P840. Light-induced Fourier transform infrared difference measurements showed that the L688C mutation induced a differential signal of the S-H stretching vibration in the P840+/P840 spectrum, as reported in P800+/P800 difference spectrum in a heliobacterial RC. Spectral changes in the 131-keto C=O region, caused by both mutations, revealed corresponding changes in the electronic structure of P840 and in the hydrogen-bonding interaction at the 131-keto C=O group. These results suggest that there is a common spatial configuration around the special pair sites among type 1 RCs. The data also provided evidence that P840 has a symmetric electronic structure, as expected from a homodimeric RC.

    DOI: 10.1038/srep19878

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    Other Link: https://www.nature.com/articles/srep19878

  • In vitro analysis of the cyanobacterial circadian oscillator composed of three Kai proteins

    K. Oyama, C. Azai, K. Terauchi

    News letter of the Japanese Society of Photosynthesis Research   25 ( 3 )   175-180 - 180   2015.12

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    Language:Japanese   Publisher:The Japanese Society of Photosynthesis Research  

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  • Menaquinone as the Secondary Electron Acceptor in the Type I Homodimeric Photosynthetic Reaction Center of Heliobacterium modesticaldum Reviewed

    Toru Kondo, Shigeru Itoh, Masahiro Matsuoka, Chihiro Azai, Hirozo Oh-oka

    The Journal of Physical Chemistry B   119 ( 27 )   8480 - 8489   2015.6

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    DOI: 10.1021/acs.jpcb.5b03723

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  • Direct observation of nitrogen fixation in filamentous cyanobacteria by using soft X-ray microscopy Invited

    T. Teramoto, M. Yoshimura, C. Azai, K. Terauchi, H. Namba, T. Ohta

    Memoirs of the SR center Ritsumeikan University   17   147 - 148   2015.5

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  • 3P103 Isolation of Rieske/cytochrome b complex from a green sulfur bacterium Chlorobaculum tepidum(03. Membrane proteins,Poster,The 52nd Annual Meeting of the Biophysical Society of Japan(BSJ2014))

    Oh-oka Hirozo, Yamamoto Kazuya, Mutoh Risa, Azai Chihiro, Kurisu Genji

    Seibutsu Butsuri   54 ( 1 )   S266   2014

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    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.54.S266_1

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  • 3P263 Two Disconnected Antenna Chlorophyll Pools in Type-1 Photosynthetic Reaction Center of Green Sulfur Bacteria(Photobiology: Photosynthesis,Poster,The 52th Annual Meeting of the Biophysical Society of Japan(BSJ2014))

    Chihiro Azai, Toru Kondo, Shigeru Itoh, Hirozo Oh-oka

    Seibutsu Butsuri   54 ( 1 )   S292   2014

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    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.54.S292_5

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  • 1P250 Structural studies on the maturation mechanism of [FeFe] hydrogenase maturation from green alga Chlamydomonas reinhardtii(18B. Photobiology:Photosynthesis,Poster,The 52nd Annual Meeting of the Biophysical Society of Japan(BSJ2014))

    Kiyota Daiki, Mutoh Risa, Azai Chihiro, Oh-oka Hirozo, Kurisu Genji

    Seibutsu Butsuri   54 ( 1 )   S182   2014

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    DOI: 10.2142/biophys.54.S182_4

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  • Molecular Construction and Reaction Mechanism of Homodimeric Photosynthetic Reaction Center

    C. Azai, H. Oh-oka

    BIOINDUSTRY   30 ( 12 )   40-47 - 47   2013.12

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    Language:Japanese  

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  • Gene Expression System in Green Sulfur Bacteria by Conjugative Plasmid Transfer Reviewed International journal

    Chihiro Azai, Jiro Harada, Hirozo Oh-oka

    PLoS ONE   8 ( 11 )   e82345 - e82345   2013.11

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Public Library of Science (PLoS)  

    DOI: 10.1371/journal.pone.0082345

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  • Site-directed mutagenesis on the photosynthetic reaction center of green sulfur bacteria

    C. Azai, H. Oh-oka

    News letter of the Japanese Society of Photosynthesis Research   23 ( 2 )   44-47 - 47   2013.8

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  • Reaction of A1-menaquinone in Type I Reaction Center of Heliobacterium modesticaldum at Cryogenic Temperature Reviewed

    Toru Kondo, Masahiro Matsuoka, Chihiro Azai, Hiroyuki Mino, Hirozo Oh-oka, Shigeru Itoh

    Advanced Topics in Science and Technology in China   21 - 24   2013

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    Language:English   Publishing type:Research paper (international conference proceedings)   Publisher:Springer Berlin Heidelberg  

    DOI: 10.1007/978-3-642-32034-7_5

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  • 3PT235 Local Structural Modifications around the Primary Electron Donor of the Green Sulfur Bacterial Photosynthetic Reaction Center(The 50th Annual Meeting of the Biophysical Society of Japan)

    Azai Chihiro, Sano Yuko, Noguchi Takumi, Oh-oka Hirozo

    Seibutsu Butsuri   52   S181   2012

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    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.52.S181_2

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  • Expression systems of exogenous gene in the obligatory anaerobic photosynthetic bacterium Chlorobaculum tepidum

    C. Azai, H. Oh-oka

    News letter of the Japanese Society of Photosynthesis Research   21 ( 3 )   95-101 - 101   2011.12

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  • A heterogeneous tag-attachment to the homodimeric type 1 photosynthetic reaction center core protein in the green sulfur bacterium Chlorobaculum tepidum Reviewed

    Chihiro Azai, Kwang Kim, Toru Kondo, Jiro Harada, Shigeru Itoh, Hirozo Oh-oka

    Biochimica et Biophysica Acta (BBA) - Bioenergetics   1807 ( 7 )   803 - 812   2011.7

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

    DOI: 10.1016/j.bbabio.2011.03.007

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  • The molecular biological method for construction of artificial heterodimeric photosynthetic reaction centers in green sulfur bacteria

    C. Azai, T. Kondo, K. Kim, J. Harada, S. Itoh, H. Oh-oka

    2011.7

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  • The photosynthetic linear electron transfer pathway in green sulfur bacteria

    Y. Tsukatani, C. Azai, H. Oh-oka

    News letter of the Japanese Society of Photosynthesis Research   20 ( 2 )   100-108 - 108   2010.8

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  • Crystal Structure of the Electron Carrier Domain of the Reaction Center Cytochrome cz Subunit from Green Photosynthetic Bacterium Chlorobium tepidum Reviewed

    Yu Hirano, Makoto Higuchi, Chihiro Azai, Hirozo Oh-oka, Kunio Miki, Zheng-Yu Wang

    Journal of Molecular Biology   397 ( 5 )   1175 - 1187   2010.4

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    DOI: 10.1016/j.jmb.2010.02.011

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  • C-type cytochromes in the photosynthetic electron transfer pathways in green sulfur bacteria and heliobacteria Invited Reviewed

    Chihiro Azai, Yusuke Tsukatani, Shigeru Itoh, Hirozo Oh-oka

    Photosynthesis Research   104 ( 2-3 )   189 - 199   2010.1

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Springer Science and Business Media LLC  

    DOI: 10.1007/s11120-009-9521-4

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  • Sulfur oxidation in mutants of the photosynthetic green sulfur bacterium Chlorobium tepidum devoid of cytochrome c-554 and SoxB Reviewed

    Chihiro Azai, Yusuke Tsukatani, Jiro Harada, Hirozo Oh-oka

    Photosynthesis Research   100 ( 2 )   57 - 65   2009.5

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    DOI: 10.1007/s11120-009-9426-2

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  • 3P-223 Spectroscopic characterization of the His-tagged homodimeric photosynthetic reaction center(Photobiology:Photosynthesis,The 47th Annual Meeting of the Biophysical Society of Japan)

    Azai Chihiro, Kondo Toru, Harada Jiro, Itoh Shigeru, Oh-oka Hirozo

    Seibutsu Butsuri   49   S188   2009

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    DOI: 10.2142/biophys.49.S188_6

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  • Accumulation of chlorophyllous pigments esterified with the geranylgeranyl group and photosynthetic competence in the CT2256-deleted mutant of the green sulfur bacterium Chlorobium tepidum Reviewed

    Jiro Harada, Shohei Miyago, Tadashi Mizoguchi, Chihiro Azai, Kazuhito Inoue, Hitoshi Tamiaki, Hirozo Oh-Oka

    Photochemical & Photobiological Sciences   7 ( 10 )   1179 - 1187   2008.10

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

    DOI: 10.1039/b802435a

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  • Parallel electron donation pathways to cytochrome cz in the type I homodimeric photosynthetic reaction center complex of Chlorobium tepidum Reviewed

    Yusuke Tsukatani, Chihiro Azai, Toru Kondo, Shigeru Itoh, Hirozo Oh-oka

    Biochimica et Biophysica Acta (BBA) - Bioenergetics   1777 ( 9 )   1211 - 1217   2008.9

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

    DOI: 10.1016/j.bbabio.2008.05.002

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  • Detection of Quinone Function in the Homodimeric Type-I Reaction Center of Heliobacterium modesticaldum Reviewed

    Toru Kondo, Hiroyuki Mino, Masahiro Matsuoka, Chihiro Azai, Hirozo Ohoka, Shigeru Itoh

    Photosynthesis. Energy from the Sun   123 - 126   2008

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    Language:English   Publishing type:Research paper (international conference proceedings)   Publisher:Springer Netherlands  

    DOI: 10.1007/978-1-4020-6709-9_28

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  • 3P-273 Arrangements of the electron transfer cofactor A_1 (menaquinone) molecule in the homodimer reaction center of Heliobacterium modesticaldum(The 46th Annual Meeting of the Biophysical Society of Japan)

    Kondo Toru, Matsuoka Masahiro, Azai Chihiro, Yoshioka Tetsunari, Mino Hiroyuki, Oh-oka Hirozo, Itoh Shigeru

    Seibutsu Butsuri   48   S169   2008

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    DOI: 10.2142/biophys.48.S169_5

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  • Bifurcated Electron Donations from Quinol Oxidoreductase and Soluble CycA to Cytochrome cz of the Photosynthetic Reaction Center Complex in the Green Sulfur Bacterium Chlorobium tepidum Reviewed

    Chihiro Azai, Yusuke Tsukatani, Ryo Miyamoto, Toru Kondo, Hiroumi Murakami, Shigeru Itoh, Hirozo Oh-oka

    Photosynthesis. Energy from the Sun   549 - 552   2008

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    DOI: 10.1007/978-1-4020-6709-9_124

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  • Electron transfer in the homodimeric reaction center core complex isolated from Heliobacterium modesticaldum Reviewed

    Kondo Toru, Miyamoto Ryo, Mino Hiroyuki, Matsuoka Masahiro, Azai Chihiro, Ohoka Hirozo, Itoh Shigeru

    Plant and Cell Physiology   48   S126   2007

  • 2P329 Arrangements of the electron transfer cofactors in the homodimer reaction center of Heliobacterium modesticaldum(Photobiology-photosynthesis,Poster Presentations)

    Kondo Toru, Matsuoka Masahiro, Azai Chihiro, Miyamoto Ryo, Mino Hiroyuki, Ohoka Hirozo, Itoh Shigeru

    Seibutsu Butsuri   47   S195   2007

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    DOI: 10.2142/biophys.47.S195_2

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  • 2P349 Fast electron donation from CycA to the homodimeric type-I reaction center of the green sulfur bacterium Chlorobium tepidum(43. Photosynthesis,Poster Session,Abstract,Meeting Program of EABS & BSJ 2006)

    Azai Chihiro, Tsukatani Yusuke, Itoh Shigeru, Oh-oka Hirozo

    Seibutsu Butsuri   46 ( 2 )   S383   2006

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    DOI: 10.2142/biophys.46.S383_1

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  • 3P278 Sulfur oxidation pathways linked with photosynthetic electron transfer pathway in the green sulfur bacterium Chlorobium tepidum

    Azai C., Tsukatani Y., Itoh S., Oh-oka H.

    Seibutsu Butsuri   45   S273   2005

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    DOI: 10.2142/biophys.45.S273_2

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▼display all

Books

  • Anoxygenic Phototrophic Bacteria

    Chihiro Azai, Jiro Harada, Akira Hiraishi, Kazuhiro Inoue, Kenji Nagashima, Keizo Shimada, Shinichi Takaichi, Yusuke Tsukatani, Zheng-Yu Wang-Otomo( Role: Contributor)

    Elsevier  2024.3  ( ISBN:9780443159824

  • 独立栄養微生物によるCO2資源化技術

    新井, 博之, 亀谷, 将史, 石井, 正治( Role: Contributor第16章 光合成によるCO2の異化的な資源化:緑色硫黄細菌を利用した実例)

    シーエムシー出版  2023.12  ( ISBN:9784781317564

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    Total pages:vii, 232p   Language:Japanese  

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  • 光合成細菌 : 酸素を出さない光合成

    嶋田, 敬三, 高市, 真一, 平石, 明, 永島, 賢治, 浅井, 智広, 原田, 二朗, 塚谷, 祐介, 王, 征宇, 井上, 和仁( Role: Contributor)

    裳華房  2020.10  ( ISBN:9784785358709

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    Total pages:xvi, 302p   Language:Japanese  

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  • 光合成のエネルギー利用と環境応用

    浅井 智広, 大岡 宏造( Role: Contributor第6章 ホモダイマー型光合成反応中心の分子構築と反応機構)

    シーエムシー出版  2014.1  ( ISBN:9784781309279

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    Total pages:305   Responsible for pages:53-61   Language:Japanese   Book type:Scholarly book

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  • 低温科学 第67巻『光合成研究法』

    原田 二朗, 浅井 智広, 大岡 宏造( Role: Contributor光合成細菌の光合成膜)

    北海道大学 低温科学研究所  2009.3 

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    Responsible for pages:205-208   Language:Japanese   Book type:Scholarly book

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    Other Link: https://eprints.lib.hokudai.ac.jp/dspace/handle/2115/39140

MISC

Presentations

  • Construction of FX-less Mutant Photosynthetic Reaction Center Complex in the Green Sulfur Bacterium Chlorobaculum tepidum

    Tomomi Inagaki, Yukie Kojima, Kazuki Terauchi, Chihiro Azai

    The 65th Annual Meeting of the Japanese Society of Plant Physiologists  ( Kobe International Conference Center (Hybrid style) )   2024.3  Japanese Society of Plant Physiologists

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    Event date: 2024.3    

    Language:English   Presentation type:Oral presentation (general)   Country:Japan  

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  • 嫌気的な生化学的操作のいろは Invited

    浅井智広

    第三回光合成タンパク質勉強会  ( オンライン )   2023.12 

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    Event date: 2023.12    

    Language:Japanese   Presentation type:Public lecture, seminar, tutorial, course, or other speech   Country:Japan  

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  • Relationship between light-harvesting function and microscopic structural heterogeneity in the photosynthetic antenna chlorosome

    Shun Arai, Tomomi Inagaki, Jiro Harada, Chihiro Azai, Toru Kondo

    The 61st Annual Meeting of the Biophysical Society of Japan  ( Nagoya Congress Center )   The Biophysical Society of Japan

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    Event date: 2023.11    

    Language:English   Presentation type:Oral presentation (general)   Country:Japan  

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  • 遺伝子から紐解く光合成とバクテリアと地球の共進化 Invited

    西原 理沙, 塚谷 祐介, 浅井 智広, 延 優

    第13回日本光合成学会年会  2023.6 

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    Event date: 2023.6    

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

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  • Electrostatic Binding of Water-soluble Subunits on the Photosynthetic Reaction Center of Green Sulfur Bacteria

    Tomomi Inagaki, Kazuki Terauchi, Chihiro Azai

    The 64th Annual Meeting of the Japanese Society of Plant Physiologists 

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    Event date: 2023.3    

    Language:Japanese   Presentation type:Oral presentation (general)   Country:Japan  

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  • Sulfur-Dependent Photoinhibition in Anaerobic Culture of Green Sulfur Bacteria

    Masahiko Higashiguchi, Kazuki Terauchi, Chihiro Azai

    The 64th Annual Meeting of the Japanese Society of Plant Physiologists  2023.3 

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    Language:Japanese   Presentation type:Oral presentation (general)   Country:Japan  

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  • Light harvesting mechanism of the smallest type-I reaction center of green sulfur bacteria: mutational modifications and theoretical analysis

    Chihiro Azai, Hirozo Oh-oka, Shigeru Itoh, Hirotaka Kito, Akihiro Kimura

    The 64th Annual Meeting of the Japanese Society of Plant Physiologists  2023.3 

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    Event date: 2023.3    

    Language:Japanese   Presentation type:Oral presentation (general)   Country:Japan  

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  • 原核生物のゲノム情報から紐解く光合成の起源と進化 Invited

    西原 亜理沙, 塚谷 祐介, 浅井 智広, 延 優

    第2回原核光合成生物シンポジウム  ( 東北大学・川内キャンパス(ハイブリッド) )   2023.3 

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    Event date: 2023.3    

    Language:Japanese   Presentation type:Oral presentation (invited, special)   Country:Japan  

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  • 緑色硫黄細菌で見つかった嫌気的な光エネルギー散逸過程 Invited

    浅井 智広

    藍藻の分子生物学2022  ( かずさアカデミアホール )   2022.12 

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    Event date: 2022.12    

    Language:Japanese   Presentation type:Oral presentation (invited, special)   Country:Japan  

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  • Revisiting the function of special pair chlorophylls in type-1 reaction centers Invited International conference

    Chihiro Azai

    The International Symposium on Photosynthesis and Chloroplast Regulation  ( Kobe Fashion Mart, Kobe )   2022.11  The Grant-in-Aid for Scientific Research on Innovative Areas, New Photosynthesis

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    Event date: 2022.11    

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

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  • 昨日の帰納はキノンの機能:タイプ1光合成反応中心にキノンは要るのか? Invited

    浅井智広

    第二回光合成タンパク質勉強会  ( オンライン )   2022.10 

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    Event date: 2022.10    

    Language:Japanese   Presentation type:Symposium, workshop panel (nominated)   Country:Japan  

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  • 緑色硫黄細菌の光合成反応中心複合体での三重項カロテノイドの生成

    浅井 智広, 木田 雅俊, 稲垣 知実, 小澄 大輔

    第34回カロテノイド研究談話会  ( 関西学院大学・西宮上ヶ原キャンパス(ハイブリッド) )   2022.9  日本カロテノイド研究会

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    Event date: 2022.9    

    Language:Japanese   Presentation type:Oral presentation (general)   Country:Japan  

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  • Charge separation induced by a lower-energy bacteriochlorophyll in the reaction center complex of Heliobacterium modesticaldum International conference

    Chihiro Azai, Risa Kojima, Kazuki Hinago, Masatoshi Kida, Tetsuya Yamamoto, Hirozo Oh-oka, Daisuke Kosumi, Yutaka Nagasawa

    The 17th International Symposium on Phototrophic Prokaryotes (ISPP2022)  ( ACC Liverpool, Liverpool )   2022.8 

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    Event date: 2022.8    

    Language:English   Presentation type:Oral presentation (general)   Country:United Kingdom  

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  • 緑色硫黄細菌Ⅰ型反応中心の励起状態の理論計算と主要BChl-a除去実験

    伊藤 繁, 木村 明洋, 鬼頭 宏任, 浅井 智広, 大岡 宏造

    第12回日本光合成学会年会  ( オンライン )   2022.5  日本光合成学会

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    Event date: 2022.5    

    Language:Japanese   Presentation type:Oral presentation (general)   Country:Japan  

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  • 緑色硫黄細菌の光合成反応中心複合体の穏和な解体

    稲垣 知実, 寺内 一姫, 浅井 智広

    第12回日本光合成学会年会  ( オンライン )   2022.5  日本光合成学会

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    Event date: 2022.5    

    Language:Japanese   Presentation type:Oral presentation (general)  

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  • 緑色硫黄細菌とメタン生成菌の人工共生系の構築 Invited

    浅井 智広

    新学術領域研究「革新的光物質変換」最終公開シンポジウム  ( 淡路夢舞台国際会議場(ハイブリッド) )   2022.3  新学術領域研究「革新的光物質変換」総括班

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    Event date: 2022.3    

    Language:Japanese   Presentation type:Oral presentation (general)   Country:Japan  

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  • Numerical Modeling for Electron Transfer Reactions in the Bacterial Type-I Photosynthetic Reaction Centers

    高橋実佳, 中庭哲津子, 武藤梨沙, 武藤梨沙, 寺内一姫, 田中秀明, 大岡宏造, 栗栖源嗣, 浅井智広

    日本植物生理学会年会(Web)  2021 

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  • Photosynthetic hydrogen production in green sulfur bacteria

    3rd International Solar Fuels Conference & International Conference on Artificial Photosynthesis-2019  2019.11 

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  • Time-Resolved Spectroscopy of Electron Transfer Processes upon Selective Excitation in the Reaction Center Complex from Heliobacterium modesticaldum

    3rd International Solar Fuels Conference & International Conference on Artificial Photosynthesis-2019  2019.11 

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  • Energy Transfer Pathways from Carotenoids in The Photosynthetic Reaction Center Complex of Green Sulfur Bacteria

    3rd International Solar Fuels Conference & International Conference on Artificial Photosynthesis-2019  2019.11 

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  • Femtosecond Time-Resolved Transient Absorption Spectroscopy of Reaction Center of Green Sulfur Bacteria: Elucidation of Primary Dynamics of Photosynthesis

    3rd International Solar Fuels Conference & International Conference on Artificial Photosynthesis-2019  2019.11 

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  • Functional implication of carotenoid glucosides in the anaerobic photosynthesis of green sulfur bacteria

    Young meeting of 3rd International Solar Fuels Conference  2019.11 

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  • Anaerobic energy dissipation by carotenoid glycoside in photosynthetic green sulfur bacteria

    1st US-Japan Binational Meeting  2019.10 

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  • Improved preparation of the reaction center complex from the green sulfur bacterium Chlorobaculum tepidum

    The 57th Annual Meeting of the Biophysical Society of Japan  2019.9 

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  • Carotenoid glycoside quenches bacteriochlorophyll a fluorescence in the photosynthetic reaction center complex of green sulfur bacteria

    The 57th Annual Meeting of the Biophysical Society of Japan  2019.9 

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  • Isolation of the Rieske/cytochrome b complex from green sulfur bacteria and interaction of the Rieske protein with cytochrome c-556

    The 57th Annual Meeting of the Biophysical Society of Japan  2019.9 

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  • 緑色硫黄細菌の光合成反応中心に結合したカロテノイドのアンテナ機能

    第33回カロテノイド研究談話会  2019.9 

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  • Functional Consequence of Glycosyl Transfer to Carotenoid in Photosynthesis of Green Sulfur Bacteria

    15th International Workshop on Supramolecular Nanoscience of Chemically Programmed Pigments (SNCPP19)  2019.6 

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  • 緑⾊硫⻩細菌のRieske/cytb複合体の精製検討とcyt c-556との相互作⽤部位の解析

    第10回 日本光合成学会年会  2019.5 

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  • 緑⾊硫⻩細菌から均⼀な反応中⼼複合体標品を得るための⼯夫

    第10回 日本光合成学会年会  2019.5 

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  • 緑⾊硫⻩細菌反応中⼼の光合成初期過程ダイナミクス

    第10回 日本光合成学会年会  2019.5 

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  • 酸素によるシアノバクテリア時計タンパク質KaiCのATPase活性阻害

    第10回 日本光合成学会年会  2019.5 

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  • 光応答性CheAタンパク質の創⽣ 〜異なる2つのクラスのヒスチジンキナーゼのキメラ化〜

    第10回 日本光合成学会年会  2019.5 

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  • カロテノイド配糖体エステルの緑⾊硫⻩細菌における光合成機能

    第10回 日本光合成学会年会  2019.5 

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  • 合成生物学時代のタンパク質の“ファイン”アート:ドメイン構造を超えたヒスチジンキナーゼのキメラ化

    圧力が拓く生命科学の新領域「圧力生命科学」研究成果報告会 〜あっと驚くタンパク質研究〜  2019.3 

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  • Primary dynamics of photosynthetic process in the reaction center of green sulfur bacteria

    The Chemical Society of Japan  2019.3 

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  • Defect of photosynthetic activity in the cruC gene mutant of the green sulfur bacterium Chlorobaculum tepidum, which lost the ability to synthesize carotenoid glucoside ester

    第60回 日本植物生理学会年会  2019.3 

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  • Interaction analysis between the Rieske/cyt b complex and c-type cytochromes in green sulfur bacteria

    第60回 日本植物生理学会年会  2019.3 

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  • Enhancement of charge separation in the photosynthetic reaction center of the green sulfur bacterium Chlorobaculum tepidum with external phylloquinones

    第60回 日本植物生理学会年会  2019.3 

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  • Creation of blue-light-responsive CheA by fusion of two different-class histidine kinases

    第60回 日本植物生理学会年会  2019.3 

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  • Circadian oscillation of cyanobacterial clock protein KaiC under anaerobic conditions

    第60回 日本植物生理学会年会  2019.3 

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  • 緑色硫黄光合成細菌におけるシアノバクテリア生物時計の細胞内再構成

    第13回日本ゲノム微生物学会年会  2019.3 

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  • 概日振動発振における時計タンパク質KaiCのC末端領域の機能解析

    第13回日本ゲノム微生物学会年会  2019.3 

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  • [FeFe]型ヒドロゲナーゼHydA1 の緑⾊硫⻩細菌Chlorobaculum tepidum細胞内での成熟化に対する鉄イオン添加の促進効果

    新学術領域研究「⾰新的光物質変換」第2回公開シンポジウム  2019.1 

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  • 緑⾊硫⻩細菌の光合成反応中⼼コアアンテナ系におけるカロテノイド配糖体エステルによる超⾼速消光

    新学術領域研究「⾰新的光物質変換」第2回公開シンポジウム  2019.1 

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  • Solubilization and purification of the Rieske/cytochrome b complex in green sulfur bacteria

    The 56th Annual Meeting of the Biophysical Society of Japan  2018.9 

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  • The orientation of menaquinone in the heliobacterial reaction center analyzed with the EPR spectroscopy

    The 56th Annual Meeting of the Biophysical Society of Japan  2018.9 

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  • Symmetry or asymmetry? - Site-specific structural modification of the homodimeric photosynthetic reaction center of green sulfur bacteria

    The 56th Annual Meeting of the Biophysical Society of Japan  2018.9 

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  • X-ray structure of the type-I reaction center from Heliobacterium modesticaldum at 3.2 Å resolution

    The 56th Annual Meeting of the Biophysical Society of Japan  2018.9 

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  • 緑色硫黄細菌Chlorobaculum tepidumのカロテノイド配糖体エステル欠損の光合成活性への影響

    第32回カロテノイド研究談話会  2018.9 

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  • Fluorescence lifetime analysis of the green sulfur bacterium Chlorobium tepidum mutant devoid of a reaction center specific carotenoid glycoside

    ISPR Conference on Microbial Photosynthesis  2018.8 

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  • Glycolipid and chlorophyll a biosyntheses are required for the heterologous assembly of the type-I photochemical reaction center in purple bacteria

    16th International Symposium on Phototrophic Prokaryotes  2018.8 

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  • Reconstitution of circadian oscillation of KaiC from the nonheterocystous nitrogen-fixing cyanobacterium Leptolyngbya boryana

    16th International Symposium on Phototrophic Prokaryotes  2018.8 

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  • BciA works as an auxiliary but highly active divinyl reductase in the purple photosynthetic bacterium Rhodobacter capsulatus

    16th International Symposium on Phototrophic Prokaryotes  2018.8 

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  • Menaquinone (MQ) in the reaction center of Heliobacterium modesticaldum studied by ESP-EPR signal of P800+MQ–: its implication in the 3D structure

    16th International Symposium on Phototrophic Prokaryotes  2018.8 

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  • Creation of phototactic CheA - Chimerization of two different classes of histidine kinases -

    14th International Workshop on Supramolecular Nanoscience of Chemically Programmed Pigments (SNCPP18)  2018.6 

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  • 緑色硫黄細菌のRieske/cyt b型シトクロム複合体の可溶化・精製の試み

    第9回 日本光合成学会年会  2018.5 

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  • 紅色光合成細菌Rhodobactor capsulatusのBciAによる迅速なクロロフィリドaのC8ビニル基還元

    第9回 日本光合成学会年会  2018.5 

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  • シアノバクテリアの時計タンパク質KaiCにおけるCII六量体リングのリン酸化と構造変化

    第59回 日本植物生理学会年会  2018.3 

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  • 異なるクラスのヒスチジンキナーゼを融合した人工青色光受容体タンパク質の作製

    第59回 日本植物生理学会年会  2018.3 

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  • 光化学系I型反応中心の紅色光合成細菌への導入

    第59回 日本植物生理学会年会  2018.3 

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  • 軟X線顕微鏡を用いた糸状性シアノバクテリアにおける光合成発生酸素の直接観察

    第59回 日本植物生理学会年会  2018.3 

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  • 光応答性CheA タンパク質の設計:異なるクラスのヒスチジンキナーゼのキメラ化

    第12回日本ゲノム微生物学会年会  2018.3 

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  • Analyses of electron transfer reactions on donor and acceptor sides in heliobacterial photosynthetic reaction center

    The 55th Annual Meeting of the Biophysical Society of Japan  2017.9 

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  • Visualization of intracellular element concentration in cyanobacteria with soft x-ray live cell imaging

    The 55th Annual Meeting of the Biophysical Society of Japan  2017.9 

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  • Conformational change by phosphorylation and ATP hydrolysis in the cyanobacterial circadian oscillator KaiC

    The 55th Annual Meeting of the Biophysical Society of Japan  2017.9 

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  • 緑色硫黄細菌Chlorobaculum tepidumの異種遺伝子発現系による、緑藻由来[FeFe]型ヒドロゲナーゼの高発現と水素生産

    第8回 日本光合成学会年会  2017.5 

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  • ヘリオバクテリア反応中心コアタンパクとFd1/Fd2の再構成実験

    第8回 日本光合成学会年会  2017.5 

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  • 分子状酸素の直接観察を目指したシアノバクテリア細胞内の元素分布の軟X線顕微鏡イメージング

    第8回 日本光合成学会年会  2017.5 

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  • 緑⾊硫⻩光合成細菌におけるシアノバクテリア時計タンパク質の強制発現

    第8回 日本光合成学会年会  2017.5 

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  • 環境の酸素レベルによるシアノバクテリア⽣物時計再構成系の周期⻑変化

    第8回 日本光合成学会年会  2017.5 

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  • シアノバクテリア時計タンパク質KaiC のCIドメインのATPase 活性に依存した2状態変換

    第58回 日本植物生理学会年会  2017.3 

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  • 軟X線顕微鏡による糸状性シアノバクテリアAnabaena sp. PCC 7120内の窒素分布の可視化

    第58回 日本植物生理学会年会  2017.3 

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  • 絶対嫌気性の光合成細菌Chlorobaculum tepidumの異種遺伝子発現系を利用した緑藻[FeFe]型ヒドロゲナーゼの細胞内成熟化

    第58回 日本植物生理学会年会  2017.3 

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  • Reconstitution experiment of the PshB protein with the photosynthetic reaction center core protein of Heliobacteriaum modesticaldum

    第58回 日本植物生理学会年会  2017.3 

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  • N-terminal nucleotide sequence dependency of anaerobic protein overexpression in the photosynthetic green sulfur bacterium Chlorobaculum tepidum

    2017 International Conference on Artificial Photosynthesis  2017.3 

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  • 窒素固定性シアノバクテリアLeptolyngbya boryanaにおける時計タンパク質KaiCの生化学的解析

    第11回 ゲノム微生物学会年会  2017.3 

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  • 光合成反応中心複合体に潜む構造機能相関の実験的探索

    第9回 BKCバイオインフォマティクス研究会  2017.2 

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  • 透過型軟X線顕微鏡によるシアノバクテリアの元素選択的な細胞内構造の観察

    PF研究会「PF挿入光源ビームラインBL-19の戦略的利用に関する研究会」  2017.1 

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  • Analyses of transient absorption changes and low-temperature fluorescence in the photosynthetic reaction center of heliobacteria

    第54回 日本生物物理学会  2016.11 

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  • Conversion between two hexameric states of KaiC for cyanobacterial circadian oscillation

    IGER International Symposium on Biological Rhythms  2016.11 

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  • Circadian Clock in the Nonheterocystous Nitrogen Fixing Cyanobacterium Leptolyngbya boryana

    4th Asian Conference on Plant-Microbe Symbiosis and Nitrogen Fixation  2016.10 

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  • シアノバクテリア時計タンパク質KaiCにおけるATP加水分解に伴う構造変換

    生物資源研究センター第1回シンポジウム  2016.9 

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  • Determination of Carbon-to-Nitrogen Ratio in the Filamentous and Heterocystous Cyanobacterium Anabaena sp. PCC 7120 with Single-cell Soft X-ray Imaging

    13th International Conference on X-ray Microscopy  2016.8 

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  • Functions of Evolutionally Conserved Antenna Chlorophylls in Type-1 Photosynthetic Reaction Centers

    Photosynthetic and Respiratory Complexes: From Structure to Function  2016.8 

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  • Mutagenic analysis of conserved antenna chlorophylls in type-1 photosynthetic reaction centers

    The 17th International Congress on Photosynthesis Research  2016.8 

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  • Orientations of FA/FB and temperature dependencies of their signal intensities in the homodimeric type 1 reaction center of Heliobacterium modesticaldum

    The 17th International Congress on Photosynthesis Research  2016.8 

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  • Structural homology around the special pair chlorophylls among type-1 reaction centers

    Satellite Meeting on Photosynthesis  2016.7 

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  • Influence of codon bias on the anaerobic protein overproduction in the phototrophic green sulfur bacterium Chlorobaculum tepidum

    Satellite Meeting on Photosynthesis  2016.7 

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  • The conformational change and protomer exchange of KaiC hexamer in the cyanobacterial circadian clock

    The 16th Annual Meeting of the Protein Science Society of Japan  2016.6 

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  • ヘリオバクテリア光合成反応中心のエネルギー移動経路に関する解析

    第7回 日本光合成学会年会  2016.5 

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  • 緑色硫黄細菌での異種遺伝子発現におけるコドン使用頻度の影響

    第7回 日本光合成学会年会  2016.5 

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  • ホモダイマー型光合成反応中心の構造と機能:光合成の古くて新しいお話

    第16回光科学若手研究会  2016.4 

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  • ヘリオバクテリア光合成反応中心の低温蛍光解析

    第57回 日本植物生理学会年会  2016.3 

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  • ヘリオバクテリア由来の1 型光合成反応中心の活性測定

    第57回 日本植物生理学会年会  2016.3 

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  • 緑色硫黄細菌Chlorobaculum tepidumにおけるGFP 発現の塩基配列依存性

    第10回 日本ゲノム微生物学会年会  2016.3 

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  • シアノバクテリア生物時計再構成系における周期長決定機構の解析

    第10回 日本ゲノム微生物学会年会  2016.3 

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  • 緑藻型[FeFe]ヒドロゲナーゼとH クラスター生合成系の緑色硫黄光合成細菌における嫌気的共発現

    第4回公開シンポジウム 新学術領域研究「人工光合成による太陽光エネルギーの物質変換:実用化に向けての異分野融合」  2016.1 

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  • 軟X線顕微鏡による糸状性シアノバクテリアの炭素窒素比の決定

    第29回 日本放射光学会年会・放射光科学合同シンポジウム  2016.1 

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  • Stability and spectroscopic characterization of heliobacterial reaction center core protein incorporated into nanoporous silica particles

    The 53rd Annual Meeting of the Biophysical Society of Japan  2015.9 

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  • Direct observation of nitrogen fixation in filamentous cyanobacteria by using soft X-ray microscopy

    The 53rd Annual Meeting of the Biophysical Society of Japan  2015.9 

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  • Construction of artificial heterodimeric photosynthetic reaction center in the green sulfur bacterium Chlorobaculum tepidum

    15th International Symposium on Phototrophic Prokaryotes  2015.8 

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  • Bacteriochlorophyll a synthesis in the green sulfur bacterium Chlorobaculum tepidum mutant lacking BchX, a subunit of chlorophyllide a oxidoreductase

    15th International Symposium on Phototrophic Prokaryotes  2015.8 

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  • A Rieske/Cytochrome b Complex in the Green Sufur Bacterium Chlorobaculum tedium

    15th International Symposium on Phototrophic Prokaryotes  2015.8 

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  • Correlation between hexameric states and phosphorylation levels of the cyanobacterial circadian clock protein KaiC

    15th International Symposium on Phototrophic Prokaryotes  2015.8 

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  • Heliobacterial type-I homodimer reaction center uses menaquinone as the secondary electron acceptor, at a little different location from that of phylloquinone in PSI

    15th International Symposium on Phototrophic Prokaryotes  2015.8 

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  • The roles of two structural states of the KaiC hexamer in the cyanobacterial clock protein complex

    第15回 日本蛋白質科学会年会  2015.6 

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  • Characterization of C8-vinyl reduction activities of BciA from Rhodobacter capsulatus

    11th International Workshop on Supramolecular Nanoscience of Chemically Programmed Pigments (SNCPP15)  2015.5 

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  • Ion exchange chromatography and gel filtration chromatography analyses of cyanobacterial clock protein KaiA

    11th International Workshop on Supramolecular Nanoscience of Chemically Programmed Pigments (SNCPP15)  2015.5 

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  • Insight into the function of alanine-422 residue of KaiC involved in autonomous synchronization of circadian phosphorylation rhythm

    11th International Workshop on Supramolecular Nanoscience of Chemically Programmed Pigments (SNCPP15)  2015.5 

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  • Temperature compensation of KaiC phosphorylation in cyanobacterial circadian clock: Biochemical analysis of two homologous domains of KaiC

    11th International Workshop on Supramolecular Nanoscience of Chemically Programmed Pigments (SNCPP15)  2015.5 

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  • ホモダイマー型光合成反応中心の分子生物学的な構造機能解析

    第6回 日本光合成学会年会およびシンポジウム  2015.5 

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  • ホモダイマー光合成反応中心複合体の人工的ヘテロダイマー化

    第62回 日本生化学会近畿支部例会  2015.5 

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  • シアノバクテリア生物時計再構成系におけるKaiCの二状態変化とATPase活性

    第62回 日本生化学会近畿支部例会  2015.5 

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  • タイプ1 光合成反応中心の保存されたアンテナクロロフィルの役割

    第56回 日本植物生理学会年会  2015.3 

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  • A cytochrome bc complexe is Rieske/cytochrome b-type in photosynthetic green sulfur bacteria

    第56回 日本植物生理学会年会  2015.3 

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  • 光合成細菌Rhodobacter capsulatusにおけるジビニル還元酵素BciAのC8ビニル基還元活性解析

    第56回 日本植物生理学会年会  2015.3 

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  • 大腸菌におけるシアノバクテリア時計遺伝子の発現とその制御解析

    第9回 日本ゲノム微生物学会年会  2015.3 

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  • 緑色硫黄細菌におけるシアノバクテリア時計遺伝子の異種発現

    第9回 日本ゲノム微生物学会年会  2015.3 

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  • 糸状シアノバクテリアの元素選択的観察

    第28回 日本放射光学会年会  2015.1 

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  • Discrimination between Two Structural States of the KaiC Hexamer in the Cyanobacterial Clock Protein Complex

    The 3rd International Symposium "Dynamical ordering of biomolecular systems for creation of integrated functions"  2015.1 

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  • Isolation of Rieske/cytochrome b complex from a green sulfur bacterium Chlorobaculum tepidum

    The 52nd Annual Meeting of the Biophysical Society of Japan  2014.9 

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  • Two Disconnected Antenna Chlorophyll Pools in Type-1 Photosynthetic Reaction Center of Green Sulfur Bacteria

    The 52nd Annual Meeting of the Biophysical Society of Japan  2014.9 

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  • Structural studies of [FeFe] hydrogenase maturation from green alga Chlamydomonas reinhardtii

    The 52nd Annual Meeting of the Biophysical Society of Japan  2014.9 

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  • Effect of redox state of KaiA on the cyanobacterial circadian clock in vitro

    10th International Workshop on Supramolecular Nanoscience of Chemically Programmed Pigments (SNCPP14)  2014.5 

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  • ヘリオバクテリアI型反応中心のキノン反応・部位はPSIと異なる

    第5回 日本光合成学会年会  2014.5 

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  • シアノバクテリア生物時計再構成系におけるKaiCの周期的構造変化解析

    第5回 日本光合成学会年会  2014.5 

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  • 緑色硫黄細菌のタイプI型RC はPS Iとは異なるエネルギー移動系をもつ

    第5回 日本光合成学会年会  2014.5 

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  • 緑色硫黄細菌Chlorobaculum tepidumにおいてbchX遺伝子はバクテリオクロロフィルa生合成に必須ではない

    第55回 日本植物生理学会年会  2014.3 

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  • 多孔質ガラス中のナノ空間における光化学系IIコア複合体からcyt c6への電子伝達

    第55回 日本植物生理学会年会  2014.3 

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  • 多孔性シリカ粒子に吸着したヘリオバクテリア反応中心コアタンパクの分光学的諸性質

    第55回 日本植物生理学会年会  2014.3 

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  • Site-specific structural modification around the special pair of homodimeric photosynthetic reaction center in green sulfur bacteria

    The 16th International Congress on Photosynthesis Research  2013.8 

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  • Studies on the photosynthetic reaction centre from the green sulfur bacterium Chlorobaculum tepidum

    The 16th International Congress on Photosynthesis Research  2013.8 

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  • Heliobacterial Reaction Center Core Complex in Nanoporous Silica Material

    The 16th International Congress on Photosynthesis Research  2013.8 

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  • Inactivation of BchX, a subunit of chlorophyllide a oxidoreductase, in the green sulfur bacterium Chlorobaculum tepidum

    Light Harvesting Satellite Meeting 2013  2013.8 

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  • Studies on the photosynthetic reaction centre from the green sulphur bacterium Chlorobaculum tepidum

    Light Harvesting Satellite Meeting 2013  2013.8 

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  • 光合成反応中心複合体における非対称な電子移動

    CREST有機太陽電池シンポジウム  2013.7 

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  • 緑色硫黄細菌の一次電子供与体P840の酸化還元差スペクトルの帰属再検討

    第4回 日本光合成学会年会  2013.5 

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  • 部位特異的変異導入によるホモダイマ―型光合成反応中心の構造改変

    第54回 日本植物生理学会年会  2013.3 

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  • 光合成反応中心の構造機能相関の解明に向けた分子生物学的なアプローチ

    第1回 立命館大学BKC 生体分子ネットワークセミナー  2013.1 

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  • 緑色硫黄細菌のホモダイマ—光合成反応中心において部位特異的変異が一次電子供与体周辺の構造に与える影響

    第50回 日本生物物理学会年会  2012.9 

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  • Studies on the reduction processes from chlorophyll a estrified with geranylgeraniol to that with ∆2,6-phytadienol in the green sulfur bacterium Chlorobaculum tepidum

    14th International Symposium on Phototrophic Prokaryotes  2012.8 

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  • The gene expression system in the Green Sulfur Bacterium Chlorobaculum tepidum by Conjugative Plasmid Transfer

    14th International Symposium on Phototrophic Prokaryotes  2012.8 

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  • 緑色硫黄細菌の光合成反応中心の部位特異的変異体:一次電子供与体P840周辺の水素結合構造の改変を例として

    第3回 日本光合成学会大会  2012.6 

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  • 人工ヘテロダイマー光合成反応中心複合体の作製と精製

    第53回 日本植物生理学会年会  2012.3 

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  • The molecular biological method for the construction of artificial heterodimeric photosynthetic reaction center in green sulfur bacteria

    The 5th Asia and oceania Conference on Photobiology  2011.8 

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  • 絶対嫌気性の光合成細菌Chlorobaculum tepidumの外来遺伝子発現系

    第2回 日本光合成学会大会  2011.6 

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  • Reaction of A1-menaquinone in type I reaction center of Heliobacterium modesticaldum at cryogenic temperature

    15th International Congress of Photosynthesis (PS2010)  2010.8 

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  • The His-tagged reaction center complex of the green sulfur bacterium Chlorobaculum tepidum

    15th International Congress of Photosynthesis (PS2010)  2010.8 

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  • 人工的ヘテロダイマー化を可能にするホモダイマー型光合成反応中心への部位特異的変異導入法

    第18回 光合成の色素系と反応中心に関するセミナー  2010.7 

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  • コアタンパク質の偽二倍体化によるホモダイマー型光合成反応中心のヘテロダイマー化

    第1回 日本光合成学会大会  2010.6 

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  • ホモダイマー型光合成反応中心のアフィニティ精製と分光学的性質

    第47回 日本生物物理学会年会  2009.10 

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  • The role of cytochrome c-554 in the electron transfer pathways in the green sulfur bacterium Chlorobium tepidum

    13th International Symposium on Phototrophic Prokaryotes  2009.8 

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  • 緑色硫黄細菌Chlorobium tepidumの光合成反応中心複合体のアフィニティクロマトグラフィによる調製

    第50回 日本植物生理学会年会  2009.3 

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  • 緑色硫黄細菌Chlorobium tepidumのホモダイマー型光化学系反応中心への変異導入法の検討

    第49回 日本植物生理学会年会  2008.3 

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  • 緑色硫黄光合成細菌の電子伝達経路の分岐とその生理的意義

    第14回 日本光生物学協会年会  2007.7 

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  • Detection of quinone function in the homodimeric type-I reaction center of Heliobacterium modesticaldum

    14th International Congress on Photosynthesis (PS2007)  2007.7 

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  • Bifurcated electron donations from quinol oxidoreductase and soluble CycA to cytochrome cz of the photosynthetic reaction center complex in the green sulfur bacterium Chlorobium tepidum

    14th International Congress on Photosynthesis (PS2007)  2007.7 

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  • 緑色硫黄細菌における反応中心への競合的な電子供与反応

    第15回 光合成の色素系と反応中心に関するセミナー  2007.6 

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  • Fast electron donation from CycA to the homodimeric type-I reaction center of the green sulfur bacteria Chlorobium tepidum

    5th East Asian Biophysics Symposium & 44th Annual Meeting of the Biophysical Society of Japan  2006.11 

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  • Involvement of CycA in thiosulfate oxidatiton in the green sulfur bacteria Chlorobium tepidum

    12th International Symposium on Phototrophic Prokaryotes  2006.8 

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  • 緑色硫黄細菌Chlorobium tepidumの光合成電子伝達経路とリンクした硫黄酸化経路

    第43回 日本生物物理学会年会  2005.11 

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  • 紅藻オキツノリ着生藻の形態的、生理的特性

    第29回 日本藻類学会大会  2005.3 

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Awards

  • The 16th International Congress on Photosynthesis Research Poster Award

    2013.8   International Society of Photosynthesis Research  

  • 第3回日本光合成学会大会 優秀発表賞

    2012.6   日本光合成学会  

  • 第2回日本光合成学会大会 優秀発表賞

    2011.6   日本光合成学会  

Research Projects

  • 光合成における超硫黄分子の貢献

    Grant number:21H05266  2021.9 - 2026.3

    日本学術振興会  科学研究費助成事業  学術変革領域研究(A)  東北大学

    石丸 泰寛, 河野 優, 浅井 智広

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    Grant amount: \81900000 ( Direct Cost: \63000000 、 Indirect Cost: \18900000 )

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  • 超高速1分子スペクトル分光法で探る生体光反応の機能的ロバスト性

    Grant number:22H02030  2022.4 - 2025.3

    日本学術振興会  科学研究費助成事業  基盤研究(B)  東京工業大学

    近藤 徹, 浅井 智広

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    Grant amount: \17550000 ( Direct Cost: \13500000 、 Indirect Cost: \4050000 )

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  • Exploring photoprotective function of carotenoid under strictly anaerobic conditions

    Grant number:21K06220  2021.4 - 2024.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)  Ritsumeikan University

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

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  • Development of photosynthetic methane production by hydrogen production in green sulfur bacteria

    Grant number:20H05119  2020.4 - 2022.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)  Ritsumeikan University

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    Grant amount: \6500000 ( Direct Cost: \5000000 、 Indirect Cost: \1500000 )

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  • Identification of pH-dependent electron transfer reactions in homodimeric reaction center complex

    Grant number:19H04734  2019.4 - 2021.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)  Ritsumeikan University

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    Grant amount: \5330000 ( Direct Cost: \4100000 、 Indirect Cost: \1230000 )

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  • Development of photosynthetic methane production by hydrogen production in green sulfur bacteria

    Grant number:18H05181  2018.4 - 2020.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)  Ritsumeikan University

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    Grant amount: \5200000 ( Direct Cost: \4000000 、 Indirect Cost: \1200000 )

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  • Unveiling proton-motive force generation by bacterial type-1 photosynthetic reaction ceters

    Grant number:17H05731  2017.4 - 2019.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)  Ritsumeikan University

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    Grant amount: \6240000 ( Direct Cost: \4800000 、 Indirect Cost: \1440000 )

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  • Construction of artificial blue light receptor for synthetic phototaxis of purple photosynthetic bacteria

    Grant number:16K14907  2016.4 - 2018.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Challenging Exploratory Research  Ritsumeikan University

    Azai Chihiro, SHIMOJI Mamiko

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    Grant amount: \3640000 ( Direct Cost: \2800000 、 Indirect Cost: \840000 )

    Through the course of this project, we tried to construct a chimera of class I and II histidine kinases (HK) to construct a photoresponsive HK that enable us switch the rotation direction of bacterial flagellar motors by light irradiation. We designed an artificial blue-light receptor protein “YC”, which is a chimera of LOV-HK and CheA, based on homology in their atomic structures. Swarming assay of the YC-expressed E. coli showed acquired phototactic response by the light-dependent autophosphorylation activity of YC. The recombinant YC protein had a photoconvertable flavin cofactor that adducts to the conserved Cys residue of the LOV domain. This is the first successful report for functional fusion of the class I and II HKs.

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  • The mechanisms to control electron transfer reactions in the homodimeric photosynthetic reaction center based on its molecular structure

    Grant number:15K07026  2015.4 - 2018.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)  Osaka University

    Oh-oka Hirozo, Itoh Shigeru, Kurisu Genji, Azai Chihiro

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    Grant amount: \5200000 ( Direct Cost: \4000000 、 Indirect Cost: \1200000 )

    Light-dependent charge separation in photosynthesis is driven by the reaction center (RC) membrane protein complex that is classified into four types; the green bacterial or heliobacterial, the purple bacterial, the photosystem (PS) I and II RCs. Compared to the latter three types of RCs, the molecular architecture of heliobacterial RC is still poorly understood, in part due to the lack of high-resolution structural information. Structure of the RC from Heliobacterium modesticaldum has been solved to a resolution of 3.2 angstrom. Although the X-ray structure, which was reported by Arizona group in 2017, contained no quinone molecule within its, our crystal structure has indicated that it was contained and seemed to function as a secondary acceptor A1. The ESP-EPR signals, which was derived from the charge separated state of P800+A1-, suggested that the quinone molecule had an almost similar orientation to but a little bit different location from that in PS I.

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  • 高い水素発生活性を実現する[FeFe]型ヒドロゲナーゼの構造デザイン

    Grant number:15H00886  2015.4 - 2017.3

    日本学術振興会  科学研究費助成事業  新学術領域研究(研究領域提案型)  立命館大学

    浅井 智広

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    Grant amount: \3120000 ( Direct Cost: \2400000 、 Indirect Cost: \720000 )

    緑藻Chlamydomonas reinhardtiiの[FeFe]ヒドロゲナーゼHydA1 は、コンパクトで有りながら非常に高い水素発生活性をもつ酵素として知られているが、活性中心であるHクラスターの形成と保持に厳密に嫌気的な環境を必要とし、生化学的な解析や応用研究が困難である。本研究課題では、酸素分圧1 ppm 以下の嫌気環境でのタンパク質生産を容易に実現できる、絶対嫌気性の光合成細菌Chlorobaculum tepidumの外来遺伝子発現系を利用し、高い水素発生活性を保持したHydA1の大量生産系を確立することを主な目的としていた。
    28年度までの研究により、C. tepidumにおいて高レベルのタンパク質発現は遺伝子のN末端領域の塩基配列に強く依存することを明らかにした。29年度はこの研究成果を利用し、Hクラスター形成に必要な成熟化タンパク質であるHydE、HydF、HydGのHydA1との高レベル共発現を試みた。全長にわたるコドン使用頻度の最適化と、N末端に付加するアフィニティータグの塩基配列を検討した結果、Strepタグ付きの遺伝子の導入によってC. tepidum内でHydEFGを高発現できることがわかった。HydA1と共発現させた結果、ほぼ全てのHydA1にHクラスターを保持させることに成功した。
    驚くべきことに、HydA1とHydEFGのC. tepidum共発現株は、培養中に光依存的に水素ガスを発生させていることがわかった。HydA1は本来、光合成伝達系で働くフェレドキシンPetFをプロトン還元の電子供与体としているが、その分子認識は厳密ではなく、C. tepidumの細胞内では別の電子供与体が機能している可能性が示唆された。

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  • Functional analysis of quinone in the photosynthetic reaction center complex of green sulfur bacteria

    Grant number:25891025  2013.8 - 2015.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Research Activity Start-up  Ritsumeikan University

    AZAI Chihiro

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    Grant amount: \2730000 ( Direct Cost: \2100000 、 Indirect Cost: \630000 )

    A gene expression system by a broad host range plasmid was established in the green sulfur bacterium Chlorobaculum tepidum, which is a strictly anaerobic photosynthetic bacterium. Large-scale preparation of the homodimeric photosynthetic reaction center (RC) complex of green sulfur bacteria was easily achieved by over-expression of the His-tagged RC core-protein and its affinity purification.
    Electron transfer reactions in the green sulfur bacterial RC complex were observed by transient absorption spectroscopy from sub-nanosecond to millisecond time domains. The electron transfer pathway was affected by additives in the reaction solution; it was suggested that there is an unidentified cofactor serving as an alternate terminal electron acceptor of the normal iron-sulfur cluster in the green sulfur bacterial RC complex.

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  • In vivo reconstitution of circadian system by heterologously expression of the cyanobacterial Kai proteins

    Grant number:23570066  2011.4 - 2015.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)  Ritsumeikan University

    TERAUCHI Kazuki, AZAI Chihiro

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    Grant amount: \5460000 ( Direct Cost: \4200000 、 Indirect Cost: \1260000 )

    The cyanobacterial circadian oscillator composed of only three proteins, KaiA, KaiB, and KaiC sustains an ordered pattern of KaiC phosphorylation. In vitro reconstitution of the KaiC phosphorylation cycle, achieved by simple mixing of the three Kai proteins with ATP provided a means of studying the detailed mechanisms of the Kai-proteins oscillator. The expressions of many genes showed the circadian rhythm in the cyanobacterium Synechococcus elongates PCC 7942. In this study, we examined whether the cyanobacterial circadian clock composed of Kai proteins operate in a heterologous host. Three kai genes of the cyanobacterium S. elongates PCC 7942 were introduced to Escherichia coli or Chlorobuim tepidum, and KaiA, KaiB, KaiC proteins were co-expressed in the cells. The regulation of the molar ratio of three Kai proteins in the cell was found to be important for the exhibition of circadian rhythm in a heterologous host.

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  • 人工的ヘテロダイマー化による光合成反応中心の電子移動非対称化機構の解析

    Grant number:11J06665  2011 - 2013

    日本学術振興会  科学研究費助成事業  特別研究員奨励費  名古屋大学

    浅井 智広

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    Grant amount: \1600000 ( Direct Cost: \1600000 )

    本研究課題は、緑色硫黄細菌Chlorobaculum(Cba.)tepidumのホモダイマー光合成反応中心(RC)を、コアタンパク質遺伝子(pscA)の偽二倍体化と部位特異的変異導入によりヘテロダイマー化し、RC内で起こる非対称な電子移動のメカニズムを明らかにすることを目指している。本年度は、2種類の変異体RCについて分光学的な解析を行った。また昨年度末の時点で難航していた人工ヘテロダイマーRCの精製方法の確立を進めた。
    昨年度までに確立したCba.tepidumの発現プラスミド発現系を利用し、P840近傍の水素結合可能な位置に存在するLeuとValの変異体RCを作製した。HisタグとStrepタグのタンデムアフィニティクロマトグラフィにより変異体ホモダイマーRCを精製し、フーリエ変換赤外分光法によりP840^+/P840の差スペクトルを解析したところ、どちらの変異体でもクロロフィル色素のケトカルボニル基の伸縮振動バンドが変化した。さらに分光電気化学的解析から、どちらの変異体RCもP840^+/P840の電位を上昇させることがわかった。これは緑色硫黄細菌のRCにおいて部特異的変異体の作製と解析に成功した初めての例である。
    一方、人工ヘテロダイマーRCの精製については、タンデムアフィニティクロマトグラフィの条件最適化を検討した。その結果、StrepタグとHisタグをもつヘテロダイマーRCのStrep-tactin固定化樹脂への吸着は塩濃度に強く依存しており、わずか100mM程度であっても回収率が劇的に低下することがわかった。
    Hisタグ精製で得られたHisタグ付きRCの画分の脱塩することにより、最適化したタンデムアフィニティ精製で分光解析に十分な収量の変異体ヘテロダイマーRCが得られることがわかった。

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  • 光合成反応中心における非対称な電子移動の発生機構の解明

    Grant number:09J01578  2009 - 2010

    日本学術振興会  科学研究費助成事業  特別研究員奨励費  大阪大学

    浅井 智広

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    Grant amount: \1400000 ( Direct Cost: \1400000 )

    本研究課題は、緑色硫黄細菌Chlorobaculum tepidumのホモダイマー型光合成反応中心(RC)を任意の部位特異的変異で人工的にヘテロダイマー化し、RCにおける非対称な電子移動反応の原理の解明を目的としている。
    前年までの研究から、「RCコアタンパク質遺伝子の偽二倍体化」によって人工ヘテロダイマー型RCを実際に創出できることがわかった。本年度は、人工ヘテロダイマー型RCの特異的な精製方法の確立を目指し、C.tepidumが元からもつpscA遺伝子をN末端にStrepタグをもつpscA遺伝子と組換えた変異株を作製した。偽二倍体化で新たに導入するpscA遺伝子にはHisタグが付加されているので、この変異株を偽二倍体化用のホストとして利用すれば、Hisタグ/Strepタグのタンデムアフィニティクロマトグラフィによって人工ヘテロダイマー型RCの特異的な精製が可能になると期待される。現在、その最適な精製条件を検討中である。また、PSIとのホモロジーモデルにより電子移動への影響が予想された24のアミノ酸残基のうち、19残基について部位特異的変異を導入したpscA遺伝子発現用DNAコンストラクトの作製を完了した。現在、pscA偽二倍体化変異株を単離中である。
    これらと並行して、Hisタグによって高純度のC.tepidum RCを大量に調製可能になったことを利用し、英国グラスゴー大学との共同研究でHisタグ付きRCの結晶化条件のスクリーニングを開始した。C.tepidum RCのX線構造解析を目指し、結晶化ロボットを利用した網羅的な条件探索を進めている。

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Allotted class

  • 2024   Graduation Thesis Ⅰ   Department

  • 2024   Graduation Thesis Ⅱ   Department

  • 2024   Applied Biology   Department

  • 2024   Plant Molecular Physiology   Department

  • 2024   Basic Biotechnology   Department

  • 2024   English for Bioscience - Advanced Course 1   Department

  • 2024   English for bioscience - advanced course 2   Department

  • 2024   English for bioscience - basic course   Department

  • 2024   bio-physiolochemistry laboratory   Department

  • 2024   Molecular Cell Biology   Graduate school

  • 2024   Doctoral Research Ⅰ   Graduate school

  • 2024   Doctoral Research Ⅱ   Graduate school

  • 2024   Doctoral Research Ⅲ   Graduate school

  • 2024   Doctoral Research Ⅳ   Graduate school

  • 2024   Doctoral Research Ⅴ   Graduate school

  • 2024   Doctoral Research Ⅵ   Graduate school

  • 2024   Special lecture on Biological Sciences   Graduate school

  • 2024   Master's Research Ⅰ   Graduate school

  • 2024   Master's Research Ⅲ   Graduate school

  • 2024   Master's Research Ⅱ   Graduate school

  • 2024   Master's Research Ⅳ   Graduate school

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Teaching Experience

  • English JP2

    Institution:Ritsumeikan University

  • 英語JP2

    Institution:立命館大学

  • Graduation Research 2

    Institution:Ritsumeikan University

  • 卒業研究2

    Institution:立命館大学

  • Experiments in Molecular Biology

    Institution:Ritsumeikan University

  • 分子生物学実験

    Institution:立命館大学

  • Experiments in Basic Biology

    Institution:Ritsumeikan University

  • 生物学基礎実験

    Institution:立命館大学

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