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PGK1 Promoter Library for the Regulation of Acetate Ester Production in Saccharomyces cerevisiae during Chinese Baijiu Fermentation

Cui, Dan-yao, Zhang, Yu, Xu, Jia, Zhang, Cui-ying, Li, Wei, Xiao, Dong-guang
Journal of agricultural and food chemistry 2018 v.66 no.28 pp. 7417-7427
Saccharomyces cerevisiae, acetate esters, acetyltransferases, alcohols, beta-galactosidase, ethyl acetate, fermentation, green fluorescent protein, mutants, polymerase chain reaction, promoter regions
Appropriate concentrations and proportion of acetate esters and higher alcohols improve the quality of Chinese Baijiu. To regulate the concentrations of acetate esters in Chinese Baijiu, we constructed a PGK1 promoter library through error-prone PCR. Then, we used an enhanced green fluorescent protein as a reporter to characterize the activities of PGK1p mutants. The PGK1p library contained 28 PGK1p mutants and spanned an activity that ranged between 0.1% and 141% of wild-type PGK1p. Seven PGK1p mutants were characterized by an additional reporter β-galactosidase and then used for the overexpression of ATF1 with BAT2 deletion in Saccharomyces cerevisiae a45. The production of ethyl acetate in strains A8, A17, A18, A27, A22, A25, A28, and AWT were 1.66-, 3.09-, 10.59-, 13.07-, 15.99-, 22.67-, 24.06-, and 27.22-fold higher than that of the parental strain. The results on alcohol acetyltransferase (AATase) activity showed that the PGK1p library precisely controlled ATF1 expression and regulated the acetate esters production.