Cloning of the lipase gene of Pseudomonas mendocina P10 and its expression in Pichia pastoris

The genomic sequence of P. mendocina P10 lipase (pml) was isolated by homologous cloning method. The result showed that full-length DNA sequence of P. mendocina P10 lipase (pml) was 801bp in size, encoding a protein of 267 amino acids with a calculated molecular mass of 29.95ku.The DNA fragment coding lipase was subcloned into the pPICZαA expression vector and the reconstructed expression plasmid pPICZαA -pml was then transformed into Pichia pastoris X33. The pml was actively expressed in the Pichia pastoris X33 and the recombinant lipase protein was secreted into the cultural broth, the maximum activity of the culture supernatant was 8U/mL, when the recombinant strain was cultured on a rotary shaker.