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- Title
Characterization of Recombinant Glyoxylate Reductase from Thermophile Thermus thermophilusHB27.
- Authors
Hiroyasu Ogino; Hitoshi Nakayama; Hideyasu China; Takuya Kawata; Noriyuki Doukyu; Masahiro Yasuda
- Abstract
A glyoxylate reductase gene from the thermophilic bacterium Thermus thermophilusHB27 (TthGR) was cloned and expressed in Escherichia colicells. The recombinant enzyme was highly purified to homogeneity and characterized. The purified TthGR showed thermostability up to 70 °C. In contrast, the maximum reaction condition was relatively mild (45 °C and pH 6.7). Although the kcatvalues against co-enzyme NADH and NADPH were similar, the Kmvalue against co-enzyme NADH was ∼18 times higher than that against NADPH. TthGR prefers NADPH rather than NADH as an electron donor. These results indicate that a phosphate group of a co-enzyme affects the binding affinity rather than the reaction efficiency, and TthGR demands appropriate amount of phosphate for a high activity. Furthermore, it was found that the half-lives of TthGR in the presence of 25% dimethyl sulfoxide and diethylene glycol were significantly longer than that in the absence of an organic solvent.
- Subjects
GLYCOLS; ENZYMES; DIMETHYL sulfoxide; ETHYLENE glycol; ESCHERICHIA coli; ORGANIC solvents; ELECTRON donor-acceptor complexes; DIETHYLENE glycol
- Publication
Biotechnology Progress, 2008, Vol 24, Issue 2, p321
- ISSN
8756-7938
- Publication type
Article
- DOI
10.1021/bp0702469