Cloning, expression and characterization of meso-2,3-butanediol dehydrogenase from Klebsiella pneumoniae

被引：24
|
作者：
Zhang, Gen Lin [1 ,2 ]
Wang, Cui Wei [1 ]
Li, Chun [1 ]
机构：
[1] Beijing Inst Technol, Sch Life Sci, Beijing 100081, Peoples R China
[2] Shihezi Univ, Key Lab Green Proc Chem Engn Xinjiang Bingtuan, Shihezi 832003, Peoples R China
来源：
BIOTECHNOLOGY LETTERS | 2012年 / 34卷 / 08期
基金：
中国国家自然科学基金;
关键词：
2,3-Butanediol dehydrogenase; Enzymatic properties; Expression; Klebsiella pneumoniae; Substrate specificity; ENHANCED 2,3-BUTANEDIOL PRODUCTION; ESCHERICHIA-COLI; GENE; PATHWAY;
D O I：
10.1007/s10529-012-0933-4
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
The budC gene encoding the meso-2,3-BDH from Klebsiella pneumoniae XJ-Li was expressed in E. coli BL21 (DE3) pLys. Hypothetical amino acid sequence alignments revealed that the enzyme belongs to the short chain dehydrogenase/reductase family. After purification and refolding, the recombinant enzyme had activities of 218 U/mg for reduction of acetoin and 66 U/mg for oxidation of meso-2,3-butanediol. Highest activities were at pH 8.0 and 9.0 respectively. These are higher than other meso-2,3-butanediol dehydrogenases from K. pneumoniae. The low K (m) value (0.65 mM) for acetoin indicated that the enzyme can easily reduce acetoin to meso-2,3-butanediol. There were no significant activities towards 2R,3R-2,3-butanediol, 1,4-butanediol and 2S,3S-2,3-butanediol, suggesting that the enzyme has a high stereospecificity for the meso-dihydric alcohol.
引用
收藏
页码：1519 / 1523
页数：5
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