Expression, purification and enzymatic characteristics of aldehyde dehydrogenase from MP688
10.7644/j.issn.1674-9960.2017.05.012
- VernacularTitle:甲基营养菌MP688甲醛脱氢酶的表达、纯化及酶活性质
- Author:
Aixia JING
;
Bo BI
;
Tong LI
;
Xianghua XIONG
;
Jianhua WANG
;
Weicai ZHANG
- Keywords:
methylobacterium extorquens;
aldehyde dehydrogenase;
cloning;
purification;
enzymatic charateristics
- From:
Military Medical Sciences
2017;41(5):381-384
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone the aldehyde dehydrogenase (adhA) gene from Methylovorus glucosotrophus and study its expression,purification and enzymatic characteristics.Methods The adhA gene was amplified and cloned to the expression vector pTIG.The AdhA was successfully expressed with induction in Escherichia coli BL21(DE3).The enzymatic characteristics were investigated by AHMT,and AdhA was purified by Ni+ exchange chromatography.Results AdhA accounted for more than 50% of the total cell proteins,and the purity was about 95%.With methanol as the substrate,the optimal pH of AdhA was 7.0,while the optimal temperature was 30℃.The enzymatic activity of purified AdhA remained about 60% when stored at room temperature for 6 days.Conclusion AdhA from MP688 is expressed in vitro,and methanol is the optimal substrate among all the substrates investigated.
