文献类型：期刊文献

英文题名：Characterization of a Novel NADPH-Dependent Oxidoreductase from Gluconobacter oxydans

作者：Chen, Minmin[1];Lin, Jinping[1];Ma, Yushu[1];Wei, Dongzhi[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, New World Inst Biotechnol, Shanghai 200237, Peoples R China

年份：2010

卷号：46

期号：2

起止页码：176

外文期刊名：MOLECULAR BIOTECHNOLOGY

收录：;EI(收录号：20104613386636);WOS:【SCI-EXPANDED(收录号:WOS:000281397500010)】；

基金：This study was financially supported by The National Key Basic Research Development Program of China (NO. 2009CB724703) and National Natural Science Foundation of China (Grant No. 20976053/B060804).

语种：英文

外文关键词：Gluconobacter oxydans; NADPH-dependent aldehyde reductase; GOX0644; Protein expression; Substrate specificity

摘要：A novel protein from Gluconobacter oxydans DSM2003 which shows 60-70% similarity with members of aldo-keto reductase (AKR) superfamily was overexpressed in Escherichia coli BL21 (DE3) and purified by one step affinity chromatography with a Ni-NTA agarose column. The recombinant protein (named GOX0644) consists of 279 amino acids with an apparent molecular mass of 32 kDa in the soluble fraction, and the gene sequence encoding the protein GOX0644 is 100% identical to the ORF of gox0644 in G. oxydans 621H (DSM2343). For a detailed analysis of its enzymatic activity, the substrate specificity of the recombinant protein GOX0644 was determined. With NADPH as a cofactor, GOX0644 exhibited better activity to aromatic aldehydes, especially o-chlorobenzaldehyde, compared to aliphatic aldehydes. It showed almost no activity toward glyceraldehyde, xylose, glucose, and ketones. The protein was unable to oxidize primary- or secondary alcohols. Based on these results, GOX0644 was defined as a novel NADPH-dependent aldehyde reductase. Kinetic parameters of the protein and the dependence of its activity on temperature and pH were also determined.