传统绍兴黄酒酿造过程中酵母菌的动态监测

Dynamic Monitoring of Saccharomyces cerevisiae during the Fermentation of Traditional Shaoxing Rice Wine

以绍兴黄酒大型生产企业加饭酒为样本,采用平板分离技术和26S r DNA D1/D2序列分析技术,对发酵醪液内酵母菌数量变化和种类组成进行动态监测。结果发现,在陶缸内进行前酵时醪液内酵母菌数量表现出"急升-稳定"的变化规律,在陶坛内进行后酵时醪液内酵母菌数量遵循"缓升-缓降"的发展特征;总的来看,头耙之后直至发酵结束的近90 d的生产周期中,醪液内酵母菌数量时有波动,生活酵母缓慢减少的总趋势未曾明显改变。绍兴加饭酒发酵醪液中存在的酵母菌均被鉴定为酿酒酵母。基于Clustal X软件进行实验菌株与模式菌株之间26S rDNA D1/D2序列的比对,醪液内酵母菌种类单一却并不单纯,加饭酒的传统生产工艺并不是纯种酿酒酵母的发酵过程,至少有3个株系酿酒酵母参与生产,其中第Ⅱ类株系可能在酿酒时发挥更加重要的作用。加饭酒是绍兴黄酒市场常见的大宗产品,跟踪整个绍兴加饭酒酿造周期,探讨发酵醪液内酵母菌变化规律,所得研究结果在传统绍兴黄酒酿造的酵母菌研究方面具有代表性;对参与黄酒生产的酵母菌进行分离鉴定并收集菌种资源,可以为最终揭示传统绍兴黄酒的酿造机理奠定基础。

The community structures of yeasts in fermented mash of Jiafan rice wine collected from some large enterprise were studied according to their changing numbers and species through plate screening techniques and 26 s r DNA D1/D2 sequence analysis technology. The results showed that the numbers of yeasts presented a changing rule of ‘rapid rise-stability’ during former fermenting in vat, and a characteristic of ‘slowly rising-slowly falling’ were shown during later fermenting in jar. On the whole, although the numbers of yeasts in fermented mash during a production cycle with nearly 90 days sometimes fluctuated, they would slowly reduce. All the yeasts in fermented mash in Jiafan rice wine were identified as Saccharomyces cerevisiae. Based on the comparison results of 26 S r DNA D1/D2 sequences between test strains and type strain, it can be concluded that the kinds of yeasts in fermented mash were single, and they were not pure. The traditional production process of Jiafan rice wine was not the fermentation of pure Saccharomyces cerevisiae.They were the fermentation of at least three strains, and strain II might play a more important role. That researches about the changing rule of yeasts numbers in fermented mash of Jiafan rice wines was representative during the fermentation of traditional Shaoxing rice wine. Yeasts participating in the brewing of Shaoxing Jiafan rice wine were separated and identified, at the same time strain resources were collected systematically. The results could be applied into basic researches for revealing the brewing mechanism of traditional Shaoxing rice wine.