GPD1过表达调控葡萄酒酵母酒精和风味物质的生成量

Regulating the Production of Ethanol and Flavor by Overexpression of GPD1 in Wine Yeast

摘要:

本研究以葡萄酒酵母WY1为出发菌株，探究GPD1基因过表达对葡萄酒酵母菌产酒精能力及风味物质的影响。利用分子生物学技术，采用PCR技术扩增葡萄酒酵母WY1中甘油-3磷酸脱氢酶基因GPD1，构建了单拷贝GPD1，两拷贝GPD1和三拷贝GPD1重组菌株。利用Real-Time PCR技术对GPD1基因表达量进行检测，与出发菌株WY1相比，单拷贝GPD1，两拷贝GPD1和三拷贝GPD1重组菌株中GPD1基因的表达水平分别提高了1.75倍、3.02倍和3.42倍。葡萄酒发酵实验数据显示，GPD1基因的过表达对乙醇的降低有显著效果，单拷贝GPD1，两拷贝GPD1和三拷贝GPD1重组菌株分别比WY1降低了8.07%，14.36%和15.34%；总高级醇含量分别减少了15.20%、17.11%和16.55%；乙酸乙酯的含量分别下降了17.63%、23.81%和27.42%，乙酸异戊酯含量分别下降了11.78%、15.87%、17.79%；甘油生成量分别提高了1.02倍、1.80倍和1.83倍。本研究表明过表达GPD1基因可以影响葡萄酒酵母产酒精的能力，同时对改善葡萄酒风味有重要意义。

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.