Gene cloning and expression of an alkaline serine protease with dehairing function from Bacillus pumilus.
A new gene (named AP gene) encoding an alkaline serine protease with dehairing function was cloned from Bacillus pumilus UN-31-C-42 and its nucleotide sequence was determined. The expression of AP gene was induced with IPTG in Escherichia coli after the mature protease region was cloned into pET15b and SDS-PAGE showed expressed product clearly, but no alkaline protease activity was detected. In order to express the AP gene in B. subtilis, a recombinant expression plasmid was constructed which contained a promoter Bp53 (also from B. pumilus), the AP gene and an E. coli-B. subtilis shuttle vector pSUGV4. This plasmid was introduced into B. subtilis WB600 and the transformant displayed the hydrolyzed zone on a milk plate. The expressed product can be easily detected with SDS-PAGE and the fermentation fluid of the transformant showed low alkaline protease activity and dehairing activity. This is the first report of a gene cloned from B. pumilus, encoding an alkaline serine protease, which can alone accomplish the whole dehairing process.
Pan
Jiao
J
College of Life Science, Sichuan University and Sichuan Key Laboratory of Molecular Biology and Biotechnology, Chengdu 610064, China.
Huang
Qing
Q
Zhang
YiZheng
Y
D000818:Animals; D001407:Bacillus / Q000201:enzymology* / Q000235:genetics*; D001412:Bacillus subtilis / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D006197:Hair; D011994:Recombinant Proteins / Q000235:genetics; D012697:Serine Endopeptidases / Q000235:genetics*; D012867:Skin; D034421:Sus scrofa; D013633:Tanning
D011994:Recombinant Proteins; D012697:Serine Endopeptidases; C032923:microbial serine proteinases