De novo design and synthesis of Candida antarctica lipase B gene and ?-factor leads to high-level expression in Pichia pastoris.

ABSTRACT: Candida antarctica lipase B (CALB) is one of the most widely used and studied enzymes in the world. In order to achieve the high-level expression of CALB in Pichia, we optimized the codons of CALB gene and ?-factor by using a de novo design and synthesis strategy. Through comparative analysis of a series of recombinants with different expression components, we found that the methanol-inducible expression recombinant carrying the codon-optimized ?-factor and mature CALB gene (pPIC9K?M-CalBM) has the highest lipase production capacity. After fermentation parameters optimization, the lipase activity and protein content of the recombinant pPIC9K?M-CalBM reached 6,100 U/mL and 3.0 g/L, respectively, in a 5-L fermentor. We believe this strategy could be of special interest due to its capacity to improve the expression level of target gene, and the Pichia transformants carrying the codon-optimized gene had great potential for the industrial-scale production of CALB lipase.

SUBMITTER: Yang JK 

PROVIDER: S-EPMC3542265 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature