[Construction of novel recombinant Escherichia coli capable of producing 1,3-propanediol].
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.
Zhang
Xiao-Mei
XM
Research Center of Industrial Microorganisms, School of Biotechnology, Southern Yangtze University, Wuxi 214036, China.
Tang
Xue-Ming
XM
Zhuge
Bin
B
Shen
Wei
W
Rao
Zhi-Ming
ZM
Fang
Hui-Ying
HY
Zhuge
Jian
J
D000332:Aerobiosis; D000426:Alcohol Dehydrogenase; D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D000449:Aldehyde Reductase / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D005818:Genetic Engineering / Q000379:methods; D007527:Isoenzymes; D011409:Propylene Glycols / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D029968:Escherichia coli Proteins; D007527:Isoenzymes; D011409:Propylene Glycols; D011994:Recombinant Proteins; C041787:1,3-propanediol; D000429:Alcohol Oxidoreductases; D000426:Alcohol Dehydrogenase; C052370:1,3-propanediol dehydrogenase; D000449:Aldehyde Reductase; C528732:YqhD protein, E coli