Disruption of PDC1 Gene to Enhance Pyruvic Acid Accumulation of Saccharomyces cerevisiae

摘要

The PDC1 gene encoding a pyruvate decarboxylase in S. cerevisiae and Kanr gene were respectively amplified by PCR using oligonucleotides which contained the restriction of NotI, EcoRI, and BglII. The two genes were digested with the same EcoRI and BglII, and linked together for inserting the Kanr gene into the PDC1 gene while the gene disruption cassette was constructed. After transformation of the linear disruption cassette (P1K) into S. cerevisiae Y2, selected transformants were checked by PCR for correct recombination. The haploid mutant Y2-1△ was obtained, which could increase the yield of pyruvic acid by 176% by shaking flask cultivation as compared with the parental strain Y2.