基于NCR效应黄酒酿造鸟氨酸氨甲酰基转移酶对氨基甲酸乙酯生成的作用及分子调控机制

To elucidate the biologically metabolic mechanism of traditional fermented foods processing is the key scientific problem that determines the product quality and safety. EC is a typical contaminant produced during the processing of chinese yellow rice wine. Due to its safety risk the presence of EC causes to heavy safety problem and it was paid more attention to. However, until now it is unknown how EC was formed concerning on the cell biology, metabolism characteristic and efficient molecular regulation strategy. The previous studies made clear that EC formation was positively regulated by intracellular OTC enzyme derived from rice wine yeast cell. As a consequence, under nitrogen catabolite stress condition that Chinese yellow rice wine brewing usually faced to as research background, in this project we plans to adopt different experimental strategies including comparative metabolomics, quantitative enzymology, transcriptomics and Western blotting to uncover regulatory effects and signals of nitrogen catabolite repression (NCR) on the key enzyme of arginine metabolism-intracellular OTC and arginine metabolites; by means of metabolomics and enzyme inhibition strategies to identify key target regulating OTC maturation and to construct a specially molecular repression target on wine yeast OTC in order to directionally control EC formation in response to NCR during the brewing of rice wine. The design and implement of this project will make clear the EC-producing mechanism and OTC-involved regulating mechanism during the vinification of Chinese yellow rice wine, and reveal microbial cell metabolic pattern affecting EC formation and its molecular repression mechanism on target EC. This research will provide a novel theory for elucidating EC production and gain an interesting strategy for industrial regulation of EC.

传统发酵食品的生物代谢机理解析是决定其品质和安全的关键科学问题。黄酒酿造过程氨基甲酸乙酯是典型性的代谢性污染因子，因其风险性给黄酒产品造成严峻的安全问题引起关注，但迄今黄酒发酵EC生成的细胞生物学、代谢学及有效的分子调控仍为空白。前期研究已明晰，黄酒发酵过程EC的生成主要受到酵母胞内OTC酶的正向调控。本项目拟以黄酒发酵所处的氮营养分解代谢胁迫(NCR)环境为背景，借助比较代谢学、定量酶学、转录组学和分子印迹技术等手段解析黄酒发酵精氨酸代谢关键酶-OTC及代谢产物对EC形成的调控作用；利用代谢组学和酶学抑制研究方法鉴定调控OTC酶成熟的核心靶标，构建特异调控酵母OTC酶的分子阻遏靶标，以达到定向控制黄酒发酵EC的形成。项目的开展可望明确我国黄酒发酵EC生成的代谢规律及精氨酸代谢酶-OTC酶介入的作用机制，揭示氮阻遏效应影响EC形成的代谢规律和靶向分子阻遏机制，具有重要的理论价值和实践意义。

迄今黄酒发酵EC生成的细胞生物学、代谢学及有效的分子调控仍为空白。前期研究已明晰，黄酒发酵过程EC的生成主要受到酵母胞内OTC酶的正向调控。本研究以黄酒发酵所处的氮营养分解代谢胁迫(NCR)环境为背景，借助比较代谢学、定量酶学、组学和分子技术等手段解析黄酒发酵精氨酸代谢关键酶-OTC及代谢产物对EC形成的调控作用，发现OTC酶具有重要的负向调控作用；利用代谢组学和酶学抑制研究方法鉴定出调控OTC酶成熟的核心靶标，构建特异调控酵母OTC酶的分子阻遏靶标和重要的调控方法，达到定向控制黄酒发酵EC的形成,在黄酒酿造中实现其EC形成的阻遏率达到45%。本项目的完成明确我国黄酒发酵EC生成的代谢规律及精氨酸代谢酶-OTC酶介入的作用机制，揭示氮阻遏效应影响EC形成的代谢规律和靶向分子阻遏机制，研究结果具有重要的理论价值和实践意义。