High-level expression of lipase from Galactomyces geotrichum mafic-0601 by codon optimization in Pichia pastoris and its application in hydrolysis of various oils.

ABSTRACT: A Galactomyces geotrichum strain with lipolytic activity was isolated and identified by the analysis of internal transcribed spacer (ITS) sequence of 18 s rDNA. Full-length lipase gene of this stain is composed of 1692 base pairs (bp) without intron, which encodes a 563-amino-acid protein. A catalytic triad (Ser²¹⁷-Glu³⁵⁴-His⁴⁶³) was found by constructing the three-dimensional structure of the lipase. In shake flasks, the lipase (LIP) catalytic activity in the supernatant of the recombinant Pichia pastoris increased 48.7% by codon optimization. LIP purified by anion exchange column showed a single protein band on 12% SDS-PAGE. The molecular weight (MW) of LIP was approximately 62 kDa. The specific activity of purified LIP reached 1257.9 U/mg.

SUBMITTER: Wang J 

PROVIDER: S-EPMC6726727 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature