| Title: | de novo design and synthesis of Candida antarctica lipase B gene and alpha-factor leads to high-level expression in Pichia pastoris |
| Author(s): | Yang JK; Liu LY; Dai JH; Li Q; |
| Address: | "College of Biological and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan, Hubei, China" |
| DOI: | 10.1371/journal.pone.0053939 |
| ISSN/ISBN: | 1932-6203 (Electronic) 1932-6203 (Linking) |
| 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 alpha-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 alpha-factor and mature CALB gene (pPIC9KalphaM-CalBM) has the highest lipase production capacity. After fermentation parameters optimization, the lipase activity and protein content of the recombinant pPIC9KalphaM-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" |
| Keywords: | "Amino Acid Sequence Base Sequence Candida/*enzymology/genetics Codon/genetics Fermentation *Fungal Proteins/biosynthesis/genetics Gene Expression Regulation, Fungal *Lipase/biosynthesis/genetics Mating Factor Methanol/pharmacology Molecular Sequence Data;" |
| Notes: | "MedlineYang, Jiang-Ke Liu, Li-Ying Dai, Jiang-Hong Li, Qin eng Research Support, Non-U.S. Gov't 2013/01/18 PLoS One. 2013; 8(1):e53939. doi: 10.1371/journal.pone.0053939. Epub 2013 Jan 10" |
