Abstract

Saccharomyces cerevisiae has good reproductive ability in both haploid and diploid forms, a pyruvate decarboxylase plays an important role in S. cerevisiae cell metabolism. In this study, pdc1 and pdc5 double knockout strains of S. cerevisiae H14-02 (MATa type) and S. cerevisiae H5-02 (MATα type) were obtained by the Cre/loxP technique. The effects of the deletion of pdc1 and pdc5 on the metabolites of the two haploid S. cerevisiae strains were consistent. In S. cerevisiae H14-02, the ethanol conversion decreased by 30.19%, the conversion of glycerol increased by 40.005%, the concentration of acetic acid decreased by 43.54%, the concentration of acetoin increased by 12.79 times, and the activity of pyruvate decarboxylase decreased by 40.91% compared to those in the original H14 strain. The original S. cerevisiae haploid strain H14 produced a small amount of acetoin but produced very little 2,3-butanediol. However, S. cerevisiae H14-02 produced 1.420 ± 0.063 g/L 2,3-BD. This study not only provides strain selection for obtaining haploid strains with a high yield of 2,3-BD but also lays a foundation for haploid S. cerevisiae to be used as a new tool for genetic research and breeding programs.

摘要

Saccharomyces cerevisiae具有良好的单倍体和二倍体繁殖能力，丙酮酸脱羧酶在S. cerevisiae细胞代谢中起重要作用。本研究通过Cre/loxP技术获得了酿酒酵母H14-02（MAT a型）和酿酒酵母H5-02（MAT α型）的pdc 1和pdc 5双基因敲除菌株。pdc 1 和pdc 5的缺失对两种单倍体酿酒酵母菌株的代谢物的影响是一致的。在酿酒酵母中H14-02，乙醇转化率降低30.19%，甘油转化率提高40.005%，乙酸浓度降低43.54%，乙偶姻浓度提高12.79倍，丙酮酸脱羧酶活性降低40.91%与原始 H14 菌株中的那些相比。最初的酿酒酵母单倍体菌株 H14 产生少量的乙偶姻，但产生的 2,3-丁二醇非常少。然而，酿酒酵母H14-02 产生 1.420 ± 0.063 g/L 2,3-BD。该研究不仅为获得高产 2,3-BD 单倍体菌株提供了菌株选择，而且为单倍体酿酒酵母用作遗传研究和育种计划的新工具奠定了基础。