title: de novo design and synthesis of Candida antarctica lipase B gene and α-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 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.
authors: College of Biological and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan, Hubei, China.
authors: Jiang-Ke
authors: JK
authors: Yang
authors: Li-Ying
authors: LY
authors: Liu
authors: Jiang-Hong
authors: JH
authors: Dai
authors: Qin
authors: Q
authors: Li
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002175:Candida / Q000201:enzymology* / Q000235:genetics; D003062:Codon / Q000235:genetics; D005285:Fermentation; D005656:Fungal Proteins / Q000096:biosynthesis / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D008049:Lipase / Q000096:biosynthesis / Q000235:genetics; D000072235:Mating Factor; D000432:Methanol / Q000494:pharmacology; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics*; D010843:Pichia / Q000201:enzymology* / Q000235:genetics
chemical_list: D003062:Codon; D005656:Fungal Proteins; D010455:Peptides; D000072235:Mating Factor; D008049:Lipase; C431349:lipase B, Candida antarctica; D000432:Methanol