Analysis of the Amino Acid Residues Involved in the Thermal Properties of the Monomeric Isocitrate Dehydrogenases of the Psychrophilic Bacterium Colwellia maris and the Mesophilic Bacterium Azotobacter vinelandii

• KURIHARA Takayuki

  Division of Biological Sciences, Graduate School of Science, Hokkaido University

• TAKADA Yasuhiro

  Department of Biological Sciences, Faculty of Science, Hokkaido University

書誌事項

タイトル別名

• Analysis of the Amino Acid Residues Involved in the Thermal Properties of the Monomeric Isocitrate Dehydrogenases of the Psychrophilic Bacterium <i>Colwellia maris</i> and the Mesophilic Bacterium <i>Azotobacter vinelandii</i>

抄録

Cold-adapted monomeric isocitrate dehydrogenase of a psychrophilic bacterium, Colwellia maris, (CmIDH) showed a high degree of amino acid sequential identity (69.5%) to a mesophilic nitrogen-fixing bacterium, Azotobacter vinelandii, (AvIDH). In this study, three Ala residues of CmIDH and the corresponding Pro residues of AvIDH were exchanged by site-directed mutagenesis, and several properties of single, double, and triple mutants of the two enzymes were investigated. The mutated CmIDHs, which replaced Ala719 with Pro, showed increased activity and elevation of the optimum temperature and thermostability for activity. In contrast, mutants of AvIDH, in which Pro717 was replaced by Ala, decreased the thermostability for activity. These results indicate that Ala719 of CmIDH and Pro717 of AvIDH are involved in thermostability. On the other hand, mutated CmIDH, in which Ala710 was replaced by Pro, and the corresponding AvIDH mutant, which replaced Pro708 with Ala, showed higher and lower specific activity than the corresponding wild-type enzymes, suggesting that Pro708 of AvIDH is involved in its high catalytic ability. Furthermore, the exchange mutations between Ala740 in CmIDH and the corresponding Pro738 in AvIDH resulted in decreased and increased thermostability for CmIDH and AvIDH activity respectively, suggesting that the native Ala740 and Pro738 residues make the enzymes thermostable and thermolabile.

収録刊行物

• Ｂｉｏｓｃｉｅｎｃｅ，　Ｂｉｏｔｅｃｈｎｏｌｏｇｙ，　ａｎｄ　Ｂｉｏｃｈｅｍｉｓｔｒｙ

  Ｂｉｏｓｃｉｅｎｃｅ，　Ｂｉｏｔｅｃｈｎｏｌｏｇｙ，　ａｎｄ　Ｂｉｏｃｈｅｍｉｓｔｒｙ 76 (12), 2242-2248, 2012

  公益社団法人 日本農芸化学会

キーワード

• isocitrate dehydrogenase
• cold-adapted enzyme
• mesophilic enzyme
• site-directed mutagenesis

詳細情報

• CRID: 1390282681456710144
• NII論文ID: 130004137890
• NII論文ID: 10031146839
• NII書誌ID: AA10824164
• DOI: 10.1271/bbb.120527
• COI: 1:STN:280:DC%2BC3s7pvVSquw%3D%3D
• ISSN: 13476947
• ISSN: 09168451
• NDL書誌ID: 024165277
• PubMed: 23221716
• Web Site: http://id.ndl.go.jp/bib/024165277
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I024165277
• Web Site: http://www.tandfonline.com/doi/pdf/10.1271/bbb.120527
• 本文言語コード: en
• データソース種別:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
• 抄録ライセンスフラグ: 使用不可