短小芽孢杆菌HZbp脂肪酶基因的克隆表达

以短小芽孢杆菌HZbp总DNA为模板以PCR的方式获得512 bp的脂肪酶基因,并在该基因的两端引入了EcoR1和Sal1的酶切位点,将该基因与大肠杆菌表达质粒pSE380连接,获得重组质粒pSE380-BPL。重组质粒转入大肠杆菌表达细胞株BL21,获得工程菌株BL21-BPL。序列分析显示所克隆的基因具有脂肪酶的保守G-X-S-X-G序列,SDS-PAGE电泳显示该脂脂肪酶的分子质量约为20 kDa。在LB培养基中,IPTG诱导浓度为1.0 mmol/L,33℃诱导培养10 h后,发酵液酶活达到8 U/mL。

Cloning and expression of Bacillus pumilus HZbp lipase gene

A Bacillus pumilus lipase gene BPL with 512 bp open reading frame was cloned from total DNA and inserted into vector pSE380 to construct a new plasmid pSE380-BPL, which was transformed into E. coli strain BL21. The recombinant strain was named as BL21-BPL. The multialignment assay of the putative amino acid revealed the enzyme had conserved peptide G-X-S-X-G sequence, SDS-PAGE analysis showed that the molecular weight of the lipase protein was about 20 kDa. The engineering strain was cultured with LB, lipase could be best induced by IPTG at 1.0 mmol/L, cultured at 33 ℃ for 10 hours, the enzyme activity was about to 8 U/mL.