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Hokkaido University Collection of Scholarly and Academic Papers >
Graduate School of Science / Faculty of Science >
Peer-reviewed Journal Articles, etc >
Analysis of amino acid residues involved in cold activity of monomeric isocitrate dehydrogenase from psychrophilic bacteria, Colwellia maris and Colwellia psychrerythraea
| Title: | Analysis of amino acid residues involved in cold activity of monomeric isocitrate dehydrogenase from psychrophilic bacteria, Colwellia maris and Colwellia psychrerythraea |
| Authors: | Yasuda, Wataru Browse this author | | Kobayashi, Miyuki Browse this author | | Takada, Yasuhiro Browse this author →KAKEN DB |
| Keywords: | Isocitrate dehydrogenase | | Cold-adapted enzyme | | Site-directed mutagenesis | | Colwellia maris | | Colwellia psychrerythraea |
| Issue Date: | Nov-2013 |
| Publisher: | Soc bioscience bioengineering japan |
| Journal Title: | Journal of bioscience and bioengineering |
| Volume: | 116 |
| Issue: | 5 |
| Start Page: | 567 |
| End Page: | 572 |
| Publisher DOI: | 10.1016/j.jbiosc.2013.05.012 |
| PMID: | 23830032 |
| Abstract: | Monomeric isocitrate dehydrogenases from psychrophilic bacteria, Colwellia maris and Colwellia psychrerythraea (CmIDH-II and CpIDH-M, respectively) are cold-adapted enzymes and show a high degree of amino acid sequential identity to each other (77%). However, maximum activity of CpIDH-M at optimum temperature is much less than that of CmIDH-II. In the C-terminal region 3 of these enzymes, which was suggested from previous study to be responsible for their distinct catalytic ability, several sequential differences of amino acid residue are present. Among them, ten amino acid residues were exchanged between them by site-directed mutagenesis and several properties of the mutated enzymes were examined in this study. The mutated enzymes of CmIDH-II substituted its Gln671, Leu724 and Phe735 residues with the corresponding residues of CpIDH-M (termed Q671K, L724Q and F735L, respectively) showed lower specific activity and thermostability for activity than the wild-type enzyme. Furthermore, the decreased specific activity was also observed in L693F. In contrast, the corresponding mutants of CpIDH-M, F693L, Q724L and L735F, showed the increased specific activity and thermostability for activity. The catalytic efficiency (K-cat/K-m) values of these mutated CmIDH-II and CpIDH-M were lower and higher than those of their wild-type IDHs, respectively. These results suggest that the Gln671, Leu693, Leu724 and Phe735 residues of CmIDH-II are important for exerting its high catalytic ability. (C) 2013, The Society for Biotechnology, Japan. All rights reserved. |
| Rights: | © 2013 The Society for Biotechnology, Japan |
| Relation: | http://www.sbj.or.jp/jbb/jbb_copyright.html |
| Type: | article (author version) |
| URI: | http://hdl.handle.net/2115/54722 |
| Appears in Collections: | 理学院・理学研究院 (Graduate School of Science / Faculty of Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 高田 泰弘
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