[Construction of an engineering strain producing alkaline pectate lyase with pHSh].
The structure gene PL from Bacillus subtilis WSHB04-02 encoding pectate lyase was amplified by PCR. The pET22b(+) vector, with leader sequence PelB, harboring PL gene was constructed. From pET22b(+) PL, the fragment of PL and leader sequence PelB was amplified by PCR together, which was transformed into E. coli JMI109. The expression of PL in E. coli JM109 was not evidently different from E. coli BL21DE3 (pET22b(+) PL) which promoter is T7. SDS-PAGE analysis showed that the molecular weight of expressed recombinant PL was about 43 kDa which was the same as calculated value. The results indicated the expression of pHsh PL in E. coli JM109, Hsh as a promoter, was satisfied and low-cost. It is significant for large-scale fermentation of pectate lyase.
Zhuge
Bin
B
Laboratory of Environmental Biotechnology, School of Biotechnology, Southern Yangtze University, China. bzhuge@yahoo.com.cn
Du
Guo-cheng
GC
Shen
Wei
W
Zhuge
Jian
J
Chen
Jian
J
D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008957:Models, Genetic; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D011133:Polysaccharide-Lyases / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics
D011133:Polysaccharide-Lyases; C023911:pectate lyase