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NADP(+)-dependent isocitrate dehydrogenase from a psychrophilic bacterium, Psychromonas marina
Title: | NADP(+)-dependent isocitrate dehydrogenase from a psychrophilic bacterium, Psychromonas marina |
Authors: | Hirota, Ryo Browse this author | Tsubouchi, Kango Browse this author | Takada, Yasuhiro Browse this author →KAKEN DB |
Keywords: | Isocitrate dehydrogenase | Cold-adapted enzyme | Chimeric enzymes | Psychrophilic bacterium | Psychromonas marina |
Issue Date: | Jul-2017 |
Publisher: | Springer |
Journal Title: | Extremophiles |
Volume: | 21 |
Issue: | 4 |
Start Page: | 711 |
End Page: | 721 |
Publisher DOI: | 10.1007/s00792-017-0936-0 |
PMID: | 28447265 |
Abstract: | The gene encoding NADP+-dependent isocitrate dehydrogenase (IDH; EC 1.1.1.42) of a psychrophilic bacterium, Psychromonas marina, was cloned and sequenced. The open reading frame of the gene encoding IDH of P. marina (PmIDH) was 2229 bp in length and corresponded to a polypeptide composed of 742 amino acids. The molecular mass of IDH was calculated as 80,426 Da. The deduced amino acid sequence of PmIDH exhibited high degrees of homology with the monomeric IDH from other bacteria such as Colwellia maris (62% identity) and Azotobacter vinelandii (AvIDH) (64%). His-tagged PmIDH overexpressed in Escherichia coli cells was purified and characterized. The optimum temperature of PmIDH activity was about 35 degrees C; however, the enzyme lost 74% of the activity after incubation for 10 min at 30 degrees C, indicating that this enzyme is thermolabile. Chimeric enzymes produced through domain swapping between PmIDH and mesophilic AvIDH were constructed and their optimum temperatures and thermostability were determined. The results suggest that regions 2 and 3, especially region 3, of the two IDHs are involved in their catalytic activities and optimum temperature and thermostability for activity. |
Rights: | The final publication is available at link.springer.com |
Type: | article (author version) |
URI: | http://hdl.handle.net/2115/70883 |
Appears in Collections: | 生命科学院・先端生命科学研究院 (Graduate School of Life Science / Faculty of Advanced Life 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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