Construction of pdc-deficient Saccharomyces cerevisiae and its pyruvic acid fermentation characteristics
LI Yi1, QIN Yan1, SHEN Naikun2, ZHU Jing1, LIANG Ge1, WANG Qingyan1*
1 (National Engineering Research Center for Non-Food Bio-refinery, State Key Laboratory of Non-Food Biomass and Enzyme Technology, Guangxi Key Laboratory of Bio-refinery, Guangxi Biomass Engineering Technology Research Center, Guangxi Academy of Sciences, 98 Daling Road, Nanning 530007, China) 2 (School of Marine Sciences and Biotechnology, Guangxi Key Laboratory of Utilization of Microbial and Botanical Resources, Guangxi University for nationalities, Nanning 530008, China)
Abstract: Pyruvic acid is an important organic acid with wide applications in polymer, cosmetic, food additive, pharmaceutical and other fields. Saccharomyces cerevisiae is considered as the best candidate microorganism for pyruvic acid production. However, pyruvic acid produced during glycolysis could be catalyzed by pyruvate decarboxylase in the cytoplasm to degrade into CO2 and acetaldehyde, resulting in the loss of carbon metabolic flux. In order to redirect more carbon metabolic flux to pyruvic acid synthesis, the deletions of the three structural genes of pyruvate decarboxylase (pdc1, pdc5 and pdc6) were performed, which significantly promoted pyruvic acid accumulation. Nevertheless, poor growth of the mutant strain XY-156 was also observed. Compared with the non-evolved strain XY-156, the evolved strain XY-156A obtained by adaptive evolution showed significant improvement in cell growth, glucose consumption and pyruvate accumulation. Fed-batch fermentation was further performed in a 2 L stirred bioreactor, and up to 105 g/L pyruvic acid was obtained within 76 h, with a productivity of 1.38 g/(L·h) and a yield of 0.5 g/g glucose. The results showed that the combination of inactivating pyruvate decarboxylase with adaptive evolution could achieve efficient pyruvic acid accumulation in S. cerevisiae, and could lay a solid foundation for the industrial production of biological pyruvic acid.
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