Regulating the Production of Ethanol and Flavor by Overexpression of GPD1 in Wine Yeast
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Abstract:
The effects of metabolism regulation of GPD1 gene on ethanol-producing capacity and flavor production in wine yeast were studied, and wine yeast WY1 was used as the parental strain. In this work, three recombinant strains WY1-GPD1, WY1-2GPD1 and WY1-3GPD1 with overexpression of one, two and three copies of GPD1 gene, respectively, were constructed using molecular biology by PCR amplification of the glyceraldehyde-3-phosphate dehydrogenase gene GPD1 in Saccharomyces uvarum WY1. The expression level of GPD1 gene in the recombinant strains and the parental strain was measured by Real-time PCR, and compared with the parental strain WY1, the expression of GPD1 in recombinant strains WY1-GPD1, WY1-2GPD1 and WY1-3GPD1 increased to 1.75 times, 3.02 times and 3.42 times, respectively. The wine fermentation experiment revealed that the overexpression of GPD1 had a significant effect on the reduction of ethanol. Compared to that of WY1, ethanol yields of the recombinant strains WY1-GPD1, WY1-2GPD1 and WY1-3GPD1 were reduced by 8.07%, 14.36% and 15.34%, respectively, while total higher alcohols were decreased by 15.20%, 17.11% and 16.55%, respectively. The contents of ethyl acetate were decreased by 17.63%, 23.81% and 27.42%, and isoamyl acetate concentrations were decreased by 11.78%, 15.87% and 17.79%, respectively. The contents of glycerol were increased to1.02 times、1.80 times and 1.83 times, respectively. This study showed that overexpression of GPD1 gene can affect the ability of wine yeast to produce ethanol and is significant to improve wine flavor.
