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

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 α-factor by using a de novo design and synthesis strategy. Through comparative analysis of a s...

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Bibliographic Details
Published in:PLoS ONE
Main Authors: Jiang-Ke Yang, Li-Ying Liu, Jiang-Hong Dai, Qin Li
Format: Article in Journal/Newspaper
Language:English
Published: Public Library of Science (PLoS) 2013
Subjects:
Medicine
R
Science
Q
Antarc*
Antarctica
Online Access:https://doi.org/10.1371/journal.pone.0053939
https://doaj.org/article/9d92cdf040f34c269fed8d375fb80b96
Description
Summary: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 α-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 α-factor and mature CALB gene (pPIC9KαM-CalBM) has the highest lipase production capacity. After fermentation parameters optimization, the lipase activity and protein content of the recombinant pPIC9KαM-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.

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