Construction of novel recombinant Escherichia coli capable of producing 1,3-propanediol.
- Author:
Xiao-Mei ZHANG
1
;
Xue-Ming TANG
;
Bin ZHUGE
;
Wei SHEN
;
Zhi-Ming RAO
;
Hui-Ying FANG
;
Jian ZHUGE
Author Information
1. Research Center of Industrial Microorganisms, School of Biotechnology, Southern Yangtze University, Wuxi 214036, China.
- Publication Type:Journal Article
- MeSH:
Aerobiosis;
Alcohol Dehydrogenase;
Alcohol Oxidoreductases;
genetics;
metabolism;
Aldehyde Reductase;
genetics;
metabolism;
Escherichia coli;
enzymology;
genetics;
Escherichia coli Proteins;
genetics;
metabolism;
Genetic Engineering;
methods;
Isoenzymes;
Propylene Glycols;
metabolism;
Recombinant Proteins;
genetics;
metabolism
- From:
Chinese Journal of Biotechnology
2005;21(5):743-747
- CountryChina
- Language:Chinese
-
Abstract:
The 1,3-propanediol oxidoreductase isoenzyme encoding gene (yqhD) from E. coli was amplified by PCR. yqhD was inserted in pEtac to yield the recombinant expression vector pEtac-yqhD. Over-expression of yqhD in E. coli JM109 was achieved with pEtac-yqhD. SDS-PAGE analysis showed an over-expressed recombinant product at about 43 kD, consistent with the molecular weight predicted from gene sequence. Compared with E. coli JM109 (pEtac), the 1,3-propanediol oxidoreductase isoenzyme activity of the recombinant E. coli (pEtac-yqhD) reached 120 u/mg protein under the induction of 1.0 mmol/L IPTG at 37 degrees C for 4 hours; at similar conditions, enzyme activity of E. coli JM109 (pEtac) was only 0.5 u/mg protein. The recombinant E. coli JM109 (pUCtac-dhaB, pEtac-yqhD) was constructed. After induction with 1.0 mmol/L IPTG, the recombinant strain could transform 50 g/L glycerol to 38 g/L 1,3-propanediol under aerobic conditions. This work demonstrated firstly that the 1,3-propanediol oxidoreductase isoenzyme could show high activity under aerobic conditions.
