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Plos One : De Novo Design and Synthesis of Candida Antarctica Lipase B Gene and Α-factor Leads to High-level Expression in Pichia Pastoris, Volume 7

By Sturtevant, Joy

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Book Id: WPLBN0003935817
Format Type: PDF eBook :
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Reproduction Date: 2015

Title: Plos One : De Novo Design and Synthesis of Candida Antarctica Lipase B Gene and Α-factor Leads to High-level Expression in Pichia Pastoris, Volume 7  
Author: Sturtevant, Joy
Volume: Volume 7
Language: English
Subject: Journals, Science, Medical Science
Collections: Periodicals: Journal and Magazine Collection
Historic
Publication Date:
Publisher: Plos

Description
Description : 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 a-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 a-factor and mature CALB gene (pPIC9KaM-CalBM) has the highest lipase production capacity. After fermentation parameters optimization, the lipase activity and protein content of the recombinant pPIC9KaM-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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