酵母菌混合培养对果酒发酵的影响及分子机理解析

The mail approach to improve the modern fruit wine products is to make full use of functional yeasts including the Saccharomyces cerevisiae and non-Saccharomyces cerevisiae. In this situation, it is necessary to understand the interaction between two kinds of yeasts under the mixed culture, it is also required to know whether the metabolites or other factors are the molecular signals to interface the gene expression and changes in the cell. It means that the molecular base of their actions should be elucidated between the cells in different strains under the mixed culture, therefore molecular mechanism of cell physiological function and its changes can be explained. In this program, the variety of gene expression and protein involved among different strains were conducted under the mixed culture or environment stress by RNA-seq transcriptome sequencing technology and proteomics, combined with the small molecular metabolites analysis by NMR methods. The gene of enzymatic reaction were compared and identified, it caused the changes of yeast metabolism, flavor and nutrient changes. The aim of this study is to elucidate the effect of environment factors on physiological function and the internal adjust mechanism in different strains under the molecular level. It is beneficial to improve the fermentation optimum and ability of cell metabolism regulation, as well as providing the theory basement of improvement of fermentation control and process efficiency. It also provides the molecular tool for design of metabolism engineering strategy of fruit wine yeast. Therefore, this program is important for practical application and theory study.

现代果酒酿造业产品质量提高的主要途径是不同功能酵母菌（酿酒酵母菌和非酿酒酵母）的挖掘和充分利用；在此背景下需要深入解析酵母菌混合培养条件下细胞间相互影响规律，探究不同菌株的代谢产物或其它因素是否作为信号分子影响细胞基因表达及变化，即酵母菌混合培养条件下菌株行为效应的分子基础需要阐明，以此揭示细胞生理功能等变化的分子机制。本项目通过RNA-seq转录组测序与蛋白质组学等分析技术，结合核磁共振技术表征小分子代谢产物等分析，研究不同酵母菌混合培养或环境胁迫条件下的差异表达基因及蛋白质表达情况，并对酵母菌风味、营养等变化相关酶基因及功能验证。研究结果可从分子水平上阐明果酒发酵不同酵母菌细胞生理功能的影响因素及内部调节机制，促进酵母菌发酵的全局优化和调控代谢能力，从而为果酒酵母菌代谢控制及提高果酒发酵效能提供理论基础，也为果酒酵母菌代谢工程策略设计提供分子生物学基础，具有重要的理论和应用价值。

本项目针对现代发酵果酒行业涉及到的重要果酒酵母菌，考察在混合培养条件下酵母菌生长相互影响及其代谢的变化规律，确定影响因素及影响规律。通过RNA-seq基因组测序与转录组和蛋白质组学等分析技术，结合小分子代谢产物等的测定，研究不同酵母菌混合培养或环境胁迫条件下的差异表达基因及相关蛋白质表达差异性，并对酵母菌风味、营养等变化相关酶基因及代谢功能的分析或验证。深入解析酵母菌混合培养条件下细胞间相互影响规律，探究不同菌株的代谢产物或其它因素是否作为影响分子引起细胞基因表达及变化，以此揭示细胞生理功能等变化的分子机制。从而为果酒酵母菌代谢控制及提高果酒发酵效能提供理论基础，也为果酒酵母菌代谢工程策略设计提供分子生物学基础；具有重要的理论和应用价值。项目发表论文12篇，其中SCI与EI收录论文8篇。