Updated on 2024/11/08

写真a

 
ISHIBASHI Matsujiro
 
Organization
Research Field in Agriculture, Agriculture, Fisheries and Veterinary Medicine Area Faculty of Agriculture Department of Agriculture Professor
Research Field in Agriculture, Agriculture, Fisheries and Veterinary Medicine Area United Graduate School of Agricultural Sciences
Title
Professor

Degree

  • 博士（農学） ( 2003.3   鹿児島大学 )

Research Interests

  • 好塩性酵素

  • 異種蛋白の発現

  • 好塩菌

Research Areas

  • Others / Others  / 蛋白化学

  • Others / Others  / 微生物学

  • Others / Others  / 遺伝子工学

Research History

  • Kagoshima University   Professor

    1998.8

  • Kagoshima University

    1998.8

Professional Memberships

  • 農芸化学会

    2015.10

  • 生物工学会

    2015.10

  • 日本海水学会

    2015.2

 

Papers

  • TOKUNAGA Masao, AKUTA Teruo, TOKUNAGA Yuhei, TOMIOKA Yui, ARAKAWA Tsutomu, ISHIBASHI Matsujiro .  Agarose Gel Electrophoresis Analysis of Chemically Cross-linked ClpB Chaperone Protein from Moderate Halophile .  Salt and Seawater Science & Technology5 ( 0 ) 22 - 26   2024Agarose Gel Electrophoresis Analysis of Chemically Cross-linked ClpB Chaperone Protein from Moderate Halophile

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    Language：English   Publisher：The Society of Sea Water Science, Japan  

    Chemical cross-linking combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is a simple way to analyze oligomerization and complex formation of proteins. However, cross-linked proteins with a large molecular mass, for example with molecular weight greater than ∼400 kDa, show slow mobility and tend to stack in loading area with poor resolution. Agarose gel electrophoresis (AGE) may be more suitable, due to a large pore size of the agarose, to analyze proteins as well as DNA fragments, especially for large macromolecules and their complexes. Here, we expressed and purified recombinant ClpB chaperone protein (homologue of eukaryotic HSP104) derived from moderate halophile and analyzed the purified protein by SDS-PAGE and SDS-AGE. Chemical cross-linking and SDS-AGE analysis clearly demonstrated hexameric assembly of ClpB subunit protein. Heat-denaturation profile of the ClpB with native-AGE analysis done without SDS suggested that ClpB protein was denatured at around 50 °C.

    DOI： 10.11457/ssst.5.0_22

  • Htwe A.Z., Yamakawa T., Ishibashi M., Tsurumaru H. .  Isolation and characterization of mung bean (Vigna radiata L.) rhizobia in Myanmar .  Symbiosis   2024

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    Language：Japanese   Publisher：Symbiosis  

    We collected soil samples from six major mung bean cropping regions in Myanmar: Sagaing, Mandalay, Nay Pyi Taw, and Magway in the tropical savanna climate zone and Bago and Yangon in the tropical monsoon climate zone. All fields grew mung bean for at least 5 years and had no history of rhizobial inoculation. Mung bean ‘Yezin-11’, a popular cultivar in Myanmar, was inoculated with soil suspensions. From the nodules formed on the roots, we isolated 55 rhizobial strains. Identification of the isolates revealed the dominant species of indigenous rhizobia in each region. We identified 53 Bradyrhizobium strains and 2 Ensifer strains. Bradyrhizobium yuanmingense was dominant in the tropical savanna zone and Bradyrhizobium sp. (B. liaoningense or B. diversitatis) and B. centrosematis were dominant in the tropical monsoon zone. Principal component analysis indicates that the dominance of B. yuanmingense in the tropical savanna zone might be due to high concentration of NO3-N and P2O5 in the soil. It also indicates that the dominance of B. centrosematis in the tropical monsoon zone might be caused by drastically low pH and high concentration of NH4 in the soil. Bradyrhizobium centrosematis YGN-M9, B. yuanmingense SGG-M3, and Bradyrhizobium sp. BGO-M5 significantly increased nodulation (nodule number and nodule dry weight), acetylene reduction activity, and shoot dry weight, respectively, relative to Ensifer terangae MDY-M6. Co-inoculation with these three strains increased nodulation significantly compared with single inoculation of BGO-M5. The characterization of mung bean rhizobia and selection of microbial inoculant candidates will be useful for the development of microbial inoculants in Myanmar.

    DOI： 10.1007/s13199-024-01013-2

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  • 3-1-4 Isolation and Characterization of Mung bean Rhizobia in Myanmar（3-1 土壌生物の生態と機能 2023年度愛媛大会） .  日本土壌肥料学会講演要旨集69 ( 0 ) 28 - 28   2023.93-1-4 Isolation and Characterization of Mung bean Rhizobia in Myanmar（3-1 土壌生物の生態と機能 2023年度愛媛大会）

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    Language：Japanese   Publisher：一般社団法人 日本土壌肥料学会  

    DOI： 10.20710/dohikouen.69.0_28_1

  • Kirishima M., Yokoyama S., Matsuo K., Hamada T., Shimokawa M., Akahane T., Sugimoto T., Tsurumaru H., Ishibashi M., Mataki Y., Ootsuka T., Nomoto M., Hayashi C., Horiguchi A., Higashi M., Tanimoto A. .  Gallbladder microbiota composition is associated with pancreaticobiliary and gallbladder cancer prognosis .  BMC Microbiology22 ( 1 ) 147   2022.12

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    Language：Japanese   Publisher：BMC Microbiology  

    Background: The microbial population of the intestinal tract and its relationship to specific diseases has been extensively studied during the past decade. However, reports characterizing the bile microbiota are rare. This study aims to investigate the microbiota composition in patients with pancreaticobiliary cancers and benign diseases by 16S rRNA gene amplicon sequencing and to evaluate its potential value as a biomarker for the cancer of the bile duct, pancreas, and gallbladder. Results: We enrolled patients who were diagnosed with cancer, cystic lesions, and inflammation of the pancreaticobiliary tract. The study cohort comprised 244 patients. We extracted microbiome-derived DNA from the bile juice in surgically resected gallbladders. The microbiome composition was not significantly different according to lesion position and cancer type in terms of alpha and beta diversity. We found a significant difference in the relative abundance of Campylobacter, Citrobacter, Leptotrichia, Enterobacter, Hungatella, Mycolicibacterium, Phyllobacterium and Sphingomonas between patients without and with lymph node metastasis. Conclusions: There was a significant association between the relative abundance of certain microbes and overall survival prognosis. These microbes showed association with good prognosis in cholangiocarcinoma, but with poor prognosis in pancreatic adenocarcinoma, and vice versa. Our findings suggest that pancreaticobiliary tract cancer patients have an altered microbiome composition, which might be a biomarker for distinguishing malignancy.

    DOI： 10.1186/s12866-022-02557-3

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  • Ura T., Kameda T., Laksmi F.A., Ishibashi M., Arakawa T., Shiraki K., Hirano A. .  Affinity of phenolic compounds for transition metal ions immobilized on cation-exchange columns .  Journal of Chromatography A1676   463277   2022.8

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    Language：Japanese   Publisher：Journal of Chromatography A  

    Immobilized metal ion affinity chromatography (IMAC) is useful in purification of histidine-tagged or histidine-rich proteins and peptides from a variety of hosts. However, phenolic compounds including polyphenols interfere with IMAC due to their high affinities for the transition metals immobilized on the column resins, which hampers the purification of proteins from plant-based host systems. In contrast to extensive knowledge of the mechanism of the interactions between phenolic compounds and transition metal ions in solution, an understanding of the interactions on the columns, where transition metal ions are immobilized on the resins, remains elusive. This study systematically investigated the affinity of phenolic compounds for transition metal ions by varying the number and position of phenolic hydroxyl groups (OH groups) and using different transition metals—Fe(II), Cu(II) and Ni(II)—on various IMACs, in which the columns were fabricated by equilibrating the cation-exchange column with transition metal solutions. It was found that the more OH groups the aromatic compounds have, the higher the affinity for transition metal ions; in particular, methyl gallate and pyrogallol were permanently bound to the IMAC column, which reflected coordinate bond formation with the transition metal ions. Importantly, the phenolic compounds showed no obvious affinity for the Ni(II)-IMAC column, in contrast to the Fe(II)- and Cu(II)-IMAC columns, whereas imidazole and histidine-tagged proteins showed evident binding to the Ni(II)-IMAC column. Ni(II)-IMAC should thus be especially effective in isolating histidine-tagged and histidine-rich species from phenolic compound-containing systems. These results indicate that the affinity between phenolic compounds and transition metal ions on the column is consistent with the results in solution. They also provide a comprehensive view for devising strategies to improve IMAC purification of target proteins and peptides from samples containing phenolic compounds.

    DOI： 10.1016/j.chroma.2022.463277

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  • Tomioka Y., Arakawa T., Akuta T., Nakagawa M., Ishibashi M. .  Analysis of proteins by agarose native gel electrophoresis in the presence of solvent additives .  International Journal of Biological Macromolecules198   26 - 36   2022.2

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    Language：Japanese   Publisher：International Journal of Biological Macromolecules  

    Solvent additives, including NaCl, arginine hydrochloride (ArgHCl), glycine and sucrose, are used to enhance protein stability or reduce protein aggregation. Here, we studied the effects of these additives on proteins using agarose native gel electrophoresis. Since these additives are used at relatively high concentration, we first confirmed that they do not interfere with the performance of the native gel electrophoresis. Agarose native gel electrophoresis showed that aggregation of bovine serum albumin (BSA) induced by heating was slightly reduced by NaCl and ArgHCl. On the contrary, glycine and sucrose had marginal effects. ArgHCl and NaCl promoted heat aggregation of monoclonal antibody (mAb), while glycine and sucrose stabilized the native mAb. Arginine methyl ester inhibited heat aggregation of lysozyme and, to a much lesser extent, BSA. These results show that agarose native gel electrophoresis can be used to analyze the effects of solvent additives on proteins subjected to heat stresses. SYPRO Orange that stains only unfolded proteins confirmed unfolded structures of soluble aggregates.

    DOI： 10.1016/j.ijbiomac.2021.12.084

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  • Ishibashi M. .  Salt Mediated Modulation of Autolysis of Thermolysin-Like Proteinase, Salilysin, Isolated from a Moderate Halophile, Chromohalobacter salexigens DSM3043 .  Protein Journal40 ( 2 ) 223 - 233   2021.4

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    Language：Japanese   Publisher：Protein Journal  

    Halophilic salilysin is first synthesized as a pro-form, which has been shown autolysis activity to process pro-region (55 amino acids long) three times to form intermediate 1 (I1), intermediate 2 (I2) and final mature (M) salilysin. The autolysis of I1- to M-form salilysin in vitro was significantly accelerated with increasing NaCl concentration up to 4 M. Strong salting-out salts, (NH4)2SO4, Na2SO4 and MgSO4, were more effective, suggesting that autolysis is enhanced by inter-molecular association or structure compaction or both. However, MgCl2, a salting-in salt, was also effective, suggesting that other mechanisms, such as charge shielding and ionic binding to this halophilic protein, operated. Autolytic cleavage at site 3 resulted in mixed formation of correctly and incorrectly processed mature forms in the absence of salt, indicating that salt affected the accuracy of autolytic cleavage reaction. Far UV circular dichroism (CD) measurements indicated that E167A pro-salilysin showed an identical CD spectrum to the wild-type mature salilysin, suggesting pro-form has a proper fold for proteolytic activity. Thermal scanning indicated that E167A pro-salilysin was more heat-stable by ~ 10 °C than mature form. The CD spectra, thermal stability and modeling structure of salilysin clearly suggested that pro-salilysin is folded to the same structure as native form and is functional for autolysis.

    DOI： 10.1007/s10930-021-09964-x

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  • Matsujiro Ishibashi· Ryoichi Tanaka· Shunsuke Yamasaki· Hiroko Tokunaga1 · Tsutomu Arakawa·Masao Tokunaga .  Salt Mediated Modulation of Autolysis of Thermolysin-Like Proteinase,　Salilysin, Isolated from a Moderate Halophile, Chromohalobacter salexigens DSM3043 .  The Protein Journal   2021.1Salt Mediated Modulation of Autolysis of Thermolysin-Like Proteinase,　Salilysin, Isolated from a Moderate Halophile, Chromohalobacter salexigens DSM3043Reviewed

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

  • Tokunaga Masao, Arakawa Tsutomu, Tokunaga Yuhei, Ishibashi Matsujiro .  Efficient Expression and Refolding of Insolubilized Halophilic Starch-Binding Domain with Aggregation-Prone Peptide .  Salt and Seawater Science & Technology2 ( 0 ) 1 - 2   2021

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    Language：English   Publisher：The Society of Sea Water Science, Japan  

    Soluble expression of halophilic starch-binding domain （SBD1） from <i>Kocuria α-</i>amylase was low due to its instability in recombinant host <i>Escherichia coli</i> cells. Here, the stable SBD1 protein was expressed efficiently with an aggregation-prone peptide tag in insoluble <i>E. coli </i> inclusion bodies （IBs）. Insoluble SBD1 protein with the tag, thus obtained, was easily solubilized with 8 M urea and refolded into an active form by simple urea dilution. The yield of active SBD1 with the tag after the solubilization / dilution method was 15 ～ 30 times higher than that of direct expression of SBD1 in soluble fraction.

    DOI： 10.11457/ssst.2.0_1

  • Tanaka R. .  Salt-enhanced processing, proteolytic activity and stability of halophilic thermolysin-like proteinase, salilysin, isolated from a moderate halophile, Chromohalobacter salexigens DSM3043 .  International Journal of Biological Macromolecules164   77 - 86   2020.12

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    Publisher：International Journal of Biological Macromolecules  

    DOI： 10.1016/j.ijbiomac.2020.07.050

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  • Fina Amreta Laksmi, Shigeki Arai, Tsutomu Arakawa, Hirohito Tsurumaru,Yoshitaka Nakamura, Budi Saksono, Masao Tokunaga, Matsujiro Ishibashi .  Expression and characterization of L-arabinose isomerase from Geobacillus stearothermophilus for improved activity under acidic condition. .  Protein Expression and Purification175   2020.11 Expression and characterization of L-arabinose isomerase from Geobacillus stearothermophilus for improved activity under acidic condition.Reviewed

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  • Laksmi F.A. .  Expression and characterization of L-arabinose isomerase from Geobacillus stearothermophilus for improved activity under acidic condition .  Protein Expression and Purification175   105692   2020.11

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    Publisher：Protein Expression and Purification  

    DOI： 10.1016/j.pep.2020.105692

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  • Nakamura Y. .  Microbial Community Analysis of Digested Liquids Exhibiting Different Methane Production Potential in Methane Fermentation of Swine Feces .  Applied Biochemistry and Biotechnology191 ( 3 ) 1140 - 1154   2020.7

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    Publisher：Applied Biochemistry and Biotechnology  

    DOI： 10.1007/s12010-020-03228-7

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  • 中村　嘉孝 .  豚糞を基質とした高効率メタン発酵に関する研究 .      2020.3豚糞を基質とした高効率メタン発酵に関する研究Reviewed

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    Language：Japanese   Publishing type：Doctoral thesis  

  • Fina Amreta Laksmi .  Study on industrial application of enzymes from extremophiles .      2020.3Study on industrial application of enzymes from extremophilesReviewed

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    Language：English   Publishing type：Doctoral thesis  

  • Laksmi F.A. .  Expression, Folding, and Activation of Halophilic Alkaline Phosphatase in Non-Halophilic Brevibacillus choshinensis .  Protein Journal39 ( 1 ) 46 - 53   2020.2

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    Publisher：Protein Journal  

    DOI： 10.1007/s10930-019-09874-z

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  • Arai S. .  Catalytic mechanism and evolutionary characteristics of thioredoxin from Halobacterium salinarum NRC-1 .  Acta Crystallographica Section D: Structural Biology76 ( Pt 1 ) 73 - 84   2020.1

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    Publisher：Acta Crystallographica Section D: Structural Biology  

    DOI： 10.1107/S2059798319015894

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  • Fina Amreta Laksmi, Hikari Imamura, Hirohito Tsurumaru, Yoshitaka Nakamura, Hiroshi Hanagata, Shigeki Arai, Masao Tokunaga, Matsujiro Ishibashi .  Expression, folding, and activation of halophilic alkaline phosphatase in non-halophilic Brevibacillus choshinensis. .  The Protein Journal39   46 - 53   2020Expression, folding, and activation of halophilic alkaline phosphatase in non-halophilic Brevibacillus choshinensis.Reviewed

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

  • SHOBUDANI MIKU, AUNGZAWHTWE, YAMAKAWA TAKEO, ISHIBASHI MATSUJIRO, TSURUMARU HIROHITO .  Mutants Disrupted in the Type Ⅲ Secretion System of Bradyrhizobium elkanii BLY3-8 Overcame Nodulation Restriction by Rj₃-genotype Soybean .  Ｍｉｃｒｏｂｅｓ　ａｎｄ　ｅｎｖｉｒｏｎｍｅｎｔｓ35 ( 2 )   2020

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    Publisher：日本微生物生態学会 / 日本土壌微生物学会 / Taiwan Society of Microbial Ecology / 植物微生物研究会 / 極限環境微生物学会  

    <p><i>Bradyrhizobium elkanii</i> BLY3-8 does not form nodules on the roots of <i>Rj</i><sub>3</sub>-genotype soybean (cultivar D-51). This is a cultivar-specific nodulation restriction. The genes <i>A6X20_40975</i> and <i>A6X20_41030</i> in strain BLY3-8 were predicted to encode the transcriptional activator and apparatus of the type III secretion system (T3SS) (the proteins TtsI and RhcJ), respectively. Mutants disrupted in these genes overcame the nodulation restriction. These results suggest that an effector injected via T3SS into <i>Rj</i><sub>3</sub>-genotype soybean is involved in nodulation restriction by <i>Rj</i><sub>3</sub>-genotype soybean.</p>

    DOI： 10.1264/jsme2.ME19151

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  • Tokunaga M. .  Insoluble expression of highly soluble halophilic metal binding protein for metal ion biosorption: Application of aggregation-prone peptide from hen egg white lysozyme .  Protein Expression and Purification156   50 - 57   2019.4

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    Publisher：Protein Expression and Purification  

    DOI： 10.1016/j.pep.2019.01.001

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  • ARAKAWA Tsutomu, TOKUNAGA Hiroko, ISHIBASHI Matsujiro, TOKUNAGA Masao .  Metal Binding Properties of His-Asp Repeat Peptide from Halophilic His-rich Metal Binding Protein Monitored by Circular Dichroism Measurement .  日本海水学会誌73 ( 3 ) 159 - 164   2019

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    Publisher：日本海水学会  

    A His-Asp repeat peptide sequence was identified in a <i><u>H</u></i>is-rich metal binding <i><u>p</u></i>rotein （HP） produced by a halophilic marine bacterium, <i>Chromohalobacter salexigens</i> DSM 3043<sup>T</sup>. A synthetic peptide, PepB20, containing 8 His-Asp repeats showed binding to an Ni-loaded metal-chelating column and elution around 150-200 mM imidazole in the presence of both low （0.15 M） and high （1.5 M） concentrations of NaCl. Addition of Ni or Zn ions at 0.1 to 1 mM resulted in changes in secondary structure of PepB20, leading to metal-induced insoluble aggregation at 1 mM. PepB20 showed different circular dichroism （CD） profiles induced by the binding of Ni or Zn ions, suggesting a formation of different secondary structures by these metal ions. We described metal binding properties and consequent structure changes of PepB20 examined by CD measurements and discuss the possible physiological importance of the His-rich metal binding insertion sequence found in HP.

    DOI： 10.11457/swsj.73.3_159

  • Fina Amreta Laksmi, Shigeki Arai, Hirohito Tsurumaru, Yoshitaka Nakamura,Budi Saksono, Masao Tokunaga, Matsujiro Ishibashi .  Improved substrate specificity for D-galactose of L-arabinose isomerase for industrial application .  Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics1866 ( 11 ) 1084 - 1091   2018.11Improved substrate specificity for D-galactose of L-arabinose isomerase for industrial applicationReviewed

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

  • Laksmi F.A. .  Improved substrate specificity for D-galactose of L-arabinose isomerase for industrial application .  Biochimica et Biophysica Acta - Proteins and Proteomics1866 ( 11 ) 1084 - 1091   2018.11

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    Publisher：Biochimica et Biophysica Acta - Proteins and Proteomics  

    DOI： 10.1016/j.bbapap.2018.09.002

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  • Mizukami M. .  Efficient production of Trastuzumab Fab antibody fragments in Brevibacillus choshinensis expression system .  Protein Expression and Purification150   109 - 118   2018.10

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    Publisher：Protein Expression and Purification  

    DOI： 10.1016/j.pep.2018.05.013

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  • Ishibashi M. .  Expression and characterization of the Renilla luciferase with the cumulative mutation .  Protein Expression and Purification145   39 - 44   2018.5

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    Publisher：Protein Expression and Purification  

    DOI： 10.1016/j.pep.2017.12.010

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  • Matsujiro Ishibashi, Ryo Kawanabe, Norie Amaba, Shigeki Arai, Fina Amreta Laksmi, Kenta Komori, Masao Tokunaga .  Expression and characterization of the Renilla luciferase with the cumulative mutation .  Protein Expression and Purification145   39 - 44   2018Expression and characterization of the Renilla luciferase with the cumulative mutationReviewed

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

  • Megumi Shigehisa, Norie Amaba, Shigeki Arai, Chisato Higashi, Ryo Kawanabe, Ayano Matsunaga, Fina Amreta Laksmi, Masao Tokunaga, Matsujiro Ishibashi .  Stabilization of luciferase from Renilla reniformis using random mutations .  Protein Engineering, Design and Selection30 ( 1 ) 7 - 13   2017.1Stabilization of luciferase from Renilla reniformis using random mutationsReviewed

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  • Shigehisa M, Amaba N, Arai S, Higashi C, Kawanabe R, Matsunaga A, Laksmi FA, Tokunaga M, Ishibashi M .  Stabilization of luciferase from Renilla reniformis using random mutations. .  Protein engineering, design & selection : PEDS30 ( 1 ) 7 - 13   2017.1Stabilization of luciferase from Renilla reniformis using random mutations.

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  • 尾辻 大樹, 黒木 未知瑠, 徳永 廣子, 山口 類, 徳永 雄平, 石橋 松二郎, 荒川 力, 徳永 正雄 .  好塩性ヒスチジン高含有金属結合タンパク質の結合容量と塩環境下での重金属回収の可能性 .  日本海水学会誌71 ( 5 ) 308 - 314   2017

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    Publisher：日本海水学会  

    好塩性海洋細菌<i>Chromohalobacter salexigens</i>由来ヒスチジン高含有金属結合タンパク質（HP）は，<i>Treponema pallidum</i>由来のTroAなどの良く研究されている他の金属結合タンパク質のアミノ酸配列と比較した場合，46アミノ酸残基からなるユニークな挿入配列（多くのヒスチジン残基を含むIS46配列）を持つという特徴がある．この研究では，Zn，Cu，NiもしくはMnを含んだ緩衝液に対してHPの平衡透析を行い，HPの金属結合容量を定量した．その結果，飽和条件下で1分子のHPに最大約10分子のZn，Cu，もしくはNiイオンが結合することが分った．HPに対する金属イオンの親和性は，Zn＞Cu＞Ni＞Mnの順であった．さらに，HPを利用した塩環境下での重金属回収の可能性を試した．His-tagを付与したHP（His-tagged HP）を固定化したSepharoseカラムを作成し，金属結合量を定量したところ1分子のHis-tagged HPあたり約9.5個のNiイオンが結合した．これは塩環境下における重金属の吸着・回収にHPが利用できる可能性を示したものと考えられる．

    DOI： 10.11457/swsj.71.5_308

  • Shigehisa M. .  Stabilization of luciferase from Renilla reniformis using random mutations .  Protein Engineering, Design and Selection30 ( 1 ) 7 - 13   2017

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    Publisher：Protein Engineering, Design and Selection  

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  • NISHIMOTO Airi, OTSUJI Daiki, TOKUNAGA Hiroko, ISHIBASHI Matsujiro, ARAKAWA Tsutomu, TOKUNAGA Masao .  Characterization of Metal Binding Protein ZP and its His-rich Peptides from a Halophilic Marine Bacterium <i>Chromohalobacter salexigens</i> .  日本海水学会誌71 ( 4 ) 252 - 253   2017

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    Publisher：日本海水学会  

    We cloned a gene, Csal-0188, from genomic DNA of <i>Chromohalobacter salexigens</i> DSM 3043<sup>T</sup>. This gene was annotated to encode Zn-binding protein（ZP）, which showed high homology with the His-rich metal binding protein（HP）we reported. His-tag-ZP was successfully expressed in <i>Escherichia coli</i> and purified. ZP was constructed with a thrombin digestion site so that His-tag could be removed. ZP and chemically synthesized oligo-peptides from the His-rich insertion sequence of ZP showed several metal binding activities. This His-rich peptide was found to bind to Ni column in the presence of 8 M urea, showing clear binding under denaturation conditions, like conventional His-tag.

    DOI： 10.11457/swsj.71.4_252

  • TOKUNAGA Masao, TOKUNAGA Hiroko, ISHIBASHI Matsujiro, ARAKAWA Tsutomu .  Detergent-tolerant Halophilic &alpha;-amylase Isolated from a Moderate Halophile <i>Nesterenkonia halobia</i> .  日本海水学会誌71 ( 3 ) 189 - 190   2017

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    Publisher：日本海水学会  

    We examined the effects of several detergents on the stability of halophilic <i>Nesterenkonia halobia</i> &alpha;-amylase（NhAmy）. NhAmy was purified to homogeneity from the culture supernatant by simple one-step dextrin affinity column chromatography. The purified enzyme was found to be highly resistant to several detergents, including sodium dodecyl sulfate, which is one of the strongest protein denaturants.

    DOI： 10.11457/swsj.71.3_189

  • 石橋松二郎,大内山愉佳,井上由紀子,岩下彩香,千貫友莉,遠城道雄,徳永正雄 .  Dioscorea japonica Thunb.由来DJ1 タンパク質のクローニングと大腸菌による発現と精製 .  応用糖質科学4 ( 3 ) 241 - 248   2014.9Dioscorea japonica Thunb.由来DJ1 タンパク質のクローニングと大腸菌による発現と精製Reviewed

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

  • Keita OYAMA, Aya HAYASHI, Takekazu KOBAYASHI, Masao TOKUNAGA and Matsujiro ISHIBASHI .  Halo-tolerant Lactic Acid Bacteria from Traditional Salted Pickle Yamagawaduke in Kagoshima. .  Bull. Soc. Sea Water Sci., Jpn.68   264 - 265   2014.8Halo-tolerant Lactic Acid Bacteria from Traditional Salted Pickle Yamagawaduke in Kagoshima.Reviewed

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  • 石橋松二郎,大内山諭佳，坂部猛仁，田中玲子，大林朋子，遠城道雄，徳永正雄 .  Dioscorea japonica Thunb.で高生産されYam Viscous Polysaccharide (YVP) 画分に含まれるDJ1・タンパク質の精製 .  応用糖質科学4 ( 2 ) 167 - 172   2014.6Dioscorea japonica Thunb.で高生産されYam Viscous Polysaccharide (YVP) 画分に含まれるDJ1・タンパク質の精製Reviewed

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

  • Ishibashi M., Arake M., Tokunaga H., Arakawa T., Tokunaga M. .  Improved expression in Escherichia coli and stability of halophilic ・B-lactamase-firefly luciferase fusion protein .  Bull. Soc. Sea Water Sci.68   341 - 343   2014Improved expression in Escherichia coli and stability of halophilic ・B-lactamase-firefly luciferase fusion proteinReviewed

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  • Matsujiro Ishibashi, Tomoe Hayashi, Chiho Yoshida, Masao Tokunaga .  Increase of salt dependence of halophilic nucleoside diphosphate kinase caused by a single amino acid substitution. .  Extremophiles17 ( 4 ) 585 - 591   2013.7Increase of salt dependence of halophilic nucleoside diphosphate kinase caused by a single amino acid substitution.Reviewed

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  • Ishibashi, M., Uchino, M., Arai, S., Kuroki, R., Arakawa, T., Tokunaga, M. .  Reduction of salt-requirement of halophilic nucleoside diphosphate kinase by engineering S-S bond. .  ABB525   47 - 52   2012.6Reduction of salt-requirement of halophilic nucleoside diphosphate kinase by engineering S-S bond.Reviewed

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  • Matsujiro Ishibashia, Keiko Ida, Shuhei Tatsuda, Tsutomu Arakawa, Masao Tokunaga .  Interaction of hexa-His tag with acidic amino acids results in facilitated refolding of halophilic nucleoside diphosphate kinase. .  Int J Biol Macromol.49   778 - 783   2011.8Interaction of hexa-His tag with acidic amino acids results in facilitated refolding of halophilic nucleoside diphosphate kinase.Reviewed

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  • Matsujiro Ishibashi, Kazuki Oda, Tsutomu Arakawa and Masao Tokunaga .  Cloning, expression, purification and activation by Na ion of halophilic alkaline phosphatase from moderate halophile Halomonas sp. 593. .  Protein Expr. Purif.76   97 - 102   2011Cloning, expression, purification and activation by Na ion of halophilic alkaline phosphatase from moderate halophile Halomonas sp. 593.Reviewed

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  • Matsujiro Ishibashi, Tatsuya Iwasa, Kouko Kumeda, Tsutomu Arakawa, Masao Tokunaga .  Effects of mutations at Gly114 on the stability and refolding of haloarchaeal nucleoside diphosphate kinase in low salt solution. .  Int J Biol Macromol.44 ( 4 ) 361 - 364   2009Effects of mutations at Gly114 on the stability and refolding of haloarchaeal nucleoside diphosphate kinase in low salt solution.Reviewed

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  • 石橋松二郎、徳永正雄 .  好塩性酵素の分子メカニズム-高度好塩菌由来ヌクレオシド二リン酸キナーゼから学ぶ-． .  生化学81 ( 12 ) 1080 - 1086   2009好塩性酵素の分子メカニズム-高度好塩菌由来ヌクレオシド二リン酸キナーゼから学ぶ-．

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  • Ishibashi, M., Tatsuda, S., Izutsu, K., Kumeda, K., Arakawa, T., Tokunaga .  A single Gly114Arg mutation stabilizes the hexameric subunit assembly and changes the substrate specificity of halo-archaeal nucleoside diphosphate kinase. .  FEBS Lett.581 ( 21 ) 4073 - 4079   2007.8A single Gly114Arg mutation stabilizes the hexameric subunit assembly and changes the substrate specificity of halo-archaeal nucleoside diphosphate kinase.Reviewed

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  • Tamada, T., Honjo, E., Maeda, Y., Okamoto, T., Ishibashi, M., Tokunaga, M., Kuroki, R .  Homodimeric crossover structure of the human GCSF-receptor signaling complex. .  Proc. Natl. Acad. Sci. USA103   3135 - 3140   2006.2Homodimeric crossover structure of the human GCSF-receptor signaling complex.Reviewed

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  • Ishibashi, M., Tsumoto, K., Ejima, D., Arakawa, T., Tokunaga, M. .  Characterization of arginine as a solvent additive .  Protein Pept. Lett.12   649 - 653   2005.10Characterization of arginine as a solvent additiveReviewed

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  • Ishibashi, M., Yamashita, S., Tokunaga, M. .  Characterization of halophilic alkaline phosphatase from Halomonas sp. 593, a moderately halophilic bacterium. .  Biosci. Biotechnol. Biochem.69   1213 - 1216   2005.6Characterization of halophilic alkaline phosphatase from Halomonas sp. 593, a moderately halophilic bacterium.Reviewed

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  • Ishibashi, M., Arakawa, T., Tokunaga, M. .  Facilitated folding and subunit assembly in Escherichia coli and in vitro of nucleoside diphosphate kinase from extremely halophilic archaeon conferred by amino-terminal extension containing hexa-His-tag. .  FEBS Lett.570   87 - 92   2004.7Facilitated folding and subunit assembly in Escherichia coli and in vitro of nucleoside diphosphate kinase from extremely halophilic archaeon conferred by amino-terminal extension containing hexa-His-tag.Reviewed

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  • Mine, S., Koshiba, T., Honjo, E., Okamoto, T., Tamada, T., Maeda, Y., Matsukura, Y., Horie, A., Ishibashi, M., Sato, M., Azuma, M., Tokunaga, M., Nitta, K., Kuroki, R. .  hermodynamic analysis of the activation mechanism of the GCSF receptor induced by ligand binding. .  Biochemistry43   2458 - 2464   2004.3hermodynamic analysis of the activation mechanism of the GCSF receptor induced by ligand binding.Reviewed

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MISC

  • Is arginine a protein-denaturant?

    Ishibashi, M., Tsumoto, K., Tokunaga, M., Ejima, D., Kita, Y., Arakawa, T.

    Protein Expr. Purif.   42   1 - 6   2005.7

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    Language：English   Publishing type：Article, review, commentary, editorial, etc. (scientific journal)  

Presentations

  • Stabilization of luciferase from Renilla reniformis  

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    Event date： 2017.3

    Language：Japanese   Presentation type：Oral presentation (general)  

  • 石橋松二郎   好塩性酵素の特性とその利用   Invited International conference

    日本農芸化学会西日本支部会 

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    Event date： 2016.9

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

  • 石橋松二郎、内野真奈美、徳永正雄   高度好塩菌由来nucleoside diphosphate kinaseの構造形成と塩の役割について   International conference

    日本農芸化学会  日本農芸化学会

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    Event date： 2014.3

    Language：English  

    Venue：東京  

    国際学会

  • 石橋 松二郎、徳永 正雄   好塩性微生物と好塩性酵素  

    日本微生物生態学会  日本微生物生態学会

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    Event date： 2013.11

    Language：Japanese  

    Venue：鹿児島  

    国内学会

  • 石橋松二郎、小田一輝、荒川 力、徳永正雄   中度好塩菌Halomonas sp. 593由来好塩性アルカリフォスファターゼのクローニング・発現・精製とNaイオンによる活性化  

    日本農芸化学会  日本農芸化学会

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    Event date： 2012.3

    Language：Japanese  

    Venue：京都  

    国内学会

  • 石橋松二郎、岩佐達也、徳永正雄   高度好塩菌由来nucleoside diphosphate kinase 変異体G114Rの解析  

    日本農芸化学会  日本農芸化学会

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    Event date： 2010.3

    Language：Japanese  

    Venue：東京  

    国内学会

  • 石橋松二郎   好塩性酵素の分子メカニズム  

    九州シンポジウム  九州シンポジウム

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    Event date： 2009.9

    Language：Japanese  

    Venue：佐賀県唐津  

    国内学会

  • 石橋松二郎   好塩性古細菌由来nucleoside diphosphate kinase (NDK) Gly114Arg変異による６量体の安定化と基質特異性の変化  

    生化学会  生化学会

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    Event date： 2007.12

    Language：Japanese  

    Venue：神奈川  

    国内学会

  • 石橋松二郎   高度好塩菌由来好塩性nucleoside diphosphate kinase (HsNDK)の結晶構造解析  

    日本農芸化学会  日本農芸化学会

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    Event date： 2007.3

    Language：Japanese  

    Venue：東京  

    国内学会

  • 石橋松二郎，山下清佳，徳永正雄   中度好塩菌由来アルカリフォスファターゼ (HaALPase)の諸性質検討  

    日本農芸化学会  日本農芸化学会

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    Event date： 2006.3

    Language：Japanese  

    Venue：京都  

    国内学会

  • Tokunaga Masao, Ishibashi Matsujiro   Electrostatic and hydrophobic interactions play a major role in the stability and refolding of halophilic proteins   International conference

    The International Society for Extremophiles  The International Society for Extremophiles

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    Event date： 2005.12

    Language：English  

    Venue：東京  

    国際学会

  • 石橋松二郎、荒川力、徳永正雄   Facilitated folding in Escherichia coli and in vitro of nucleoside diphosphate kinase from extremely halophilic archaeon conferred by amino-terminal extension containing hexa-His-tag  

    日本生化学会  日本生化学会

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    Event date： 2004.10

    Language：Japanese  

    Venue：神奈川  

    国内学会

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Intellectual Property

  • 安定性が増加したウミシイタケ(Renilla reniformis)由来ルシフェラーゼ変異体

    石橋松二郎, 徳永正雄

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    Patent/Registration no：特許第6688499号  Date registered：2020.4 

    Country of applicant：Domestic