Pyruvate Decarboxylase Gene (pdc1) of Haploid Saccharomyces Cerevisiae: Knockout and Identification

doi:10.11924/j.issn.1000-6850.casb20190600332

Abstract

Abstract:

The aim is to inhibit the synthesis of ethanol, a by-product of haploid Saccharomyces cerevisiae H14, and increase the yield of 2,3-butanediol. The vector pWCL-pdc1 was constructed by genetic engineering to obtain the homologous recombinant fragment loxP-kanMX-loxP containing 40 bp pdc1 at both ends. The pdc1 deleted strain S. cerevisiae H14-01 (Δpdc1) was obtained by Cre/loxP technique. The shaking flask fermentation experiment was carried out with the wild type strain S. cerevisiae H14 as the control. The growth of S. cerevisiae H14-01 was significantly lower than that of the original strain. During the whole fermentation period, the maximum yield and conversion rate of 2,3-butanediol were 0.373 ±0.016 g/L and 0.005 g/g, respectively, which were 37.30% and 4.66% higher than that of the original strain, respectively. However, acetoin and 2,3-BD production was not detect in the original strain. In addition, the ethanol conversion of S. cerevisiae H14-01 decreased by 33.24%, but the glycerol production increased by 15.76%. It showed that after the carbon flow to ethanol was blocked, the production of 2, 3-butanediol could be increased, and part of the carbon flow will flow to glycerol. Therefore, the study lays a good foundation for further obtaining an engineering microbial population with high yield of 2,3-butanediol.

Key words: Saccharomyces cerevisiae, 2,3-butanediol, Cre/loxP technology, pyruvate decarboxylase, gene knockout

CLC Number:

Q933

Kang Jie, Wang Changli, Ge Jingping. Pyruvate Decarboxylase Gene (pdc1) of Haploid Saccharomyces Cerevisiae: Knockout and Identification[J]. Chinese Agricultural Science Bulletin, 2020, 36(24): 91-98.

Figures/Tables 12

References 31

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