Alteration of coenzyme specificity of malate dehydrogenase from Streptomyces coelicolor A3(2) by site-directed mutagenesis

被引：5
|
作者：
Ge, Y. D. [1 ]
Song, P. [1 ]
Cao, Z. Y. [1 ]
Wang, P. [1 ]
Zhu, G. P. [1 ]
机构：
[1] Anhui Normal Univ, Key Lab Biot Environm & Ecol Safety Anhui Prov, Key Lab Mol Evolut & Biodivers, Inst Mol Biol & Biotechnol, Wuhu, Anhui, Peoples R China
来源：
GENETICS AND MOLECULAR RESEARCH | 2014年 / 13卷 / 03期
基金：
中国国家自然科学基金; 高等学校博士学科点专项科研基金;
关键词：
Malate dehydrogenase; Coenzyme specificity; Streptomyces coelicolor; Site-directed mutagenesis; LACTATE-DEHYDROGENASE; XYLITOL DEHYDROGENASE; COMPLETE REVERSAL; HIGHER-PLANTS; EVOLUTION; PROTEIN; IDENTIFICATION; NADP(+); ENZYMES;
D O I：
10.4238/2014.July.29.3
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
We describe here for the first time the alteration of coenzyme specificity of malate dehydrogenase (MDH) from Streptomyces coelicolor A3(2) (ScMDH). In the present study, we replaced four amino acid residues in the Rossmann fold (beta B-alpha C) region of NADH-dependent ScMDH by site-directed mutagenesis with those of NADPH-dependent MDH (Glu42Gly, Ile43Ser, Pro45Arg, and Ala46Ser). The coenzyme specificity of the mutant enzyme (ScMDH-T4) was examined. Coenzyme specificity of ScMDH-T4 was shifted 2231.3-fold toward NADPH using k(cat)/K-m(coenzyme) as the measurement of coenzyme specificity. Accordingly, the effect of the replacements on coenzyme specificity is discussed. Our work provides further insight into the coenzyme specificity of ScMDH.
引用
收藏
页码：5758 / 5766
页数：9
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