红曲菌中嗜氮酮型和萘酮型黄色素生物合成的研究

红曲色素（Monascus pigment，MP）是红曲菌（Monascus spp., 又称红曲霉）产生的我国和东南亚地区的传统食品着色剂，已有上千年应用历史。MP是由红、橙、黄三类色素成分组成的混合物，其中黄色MP不仅种类多，且除着色功能外还有多种生物活性，具有广阔的应用前景。但目前黄色MP生物合成途径仍存在争议。申请人前期研究中发现黄色MP分为嗜氮酮型和萘酮型两类，并根据基因敲除、体外酶反应以及黄色MP产物分析等实验结果，推导了这两类黄色MP的生物合成过程，为本项目奠定了基础。据此，本申请项目拟通过组学联用的方法推导参与黄色MP合成的基因，利用基因敲除和体外酶反应方法表征酶的功能，通过构建体外多酶共催化体系和多基因异源共表达菌株来验证其生物合成过程。本项目不仅能够解析两类黄色MP的生物合成机制，为黄色MP生产菌株的改良奠定理论基础，也为其他真菌类似代谢产物合成途径的研究提供借鉴意义。

Monascus pigment (MP), produced by Monascus spp. as a traditional food colorant in China and Southeast Asia, has been utilized for more than thousands of years. MP is the mixture of red, orange and yellow pigment constituents. Among them, yellow MP has wide application prospects because of its structure diversity, good coloring effect and various biological functions. However, the biosynthetic pathway of yellow MP is still a debatable question. In our previous research, yellow MP was classified into two kinds: azaphilone and naphthalenone. Based on the results of gene knock-out, enzymatic reaction in vitro, and yellow MP metabolites analysis, the possible biosynthetic pathway was proposed. This research will firstly search the genes involved in biosynthesis through metabolomics-proteomics analysis. Then, the catalytic function of those enzymes will be determined by gene knock-out and in vitro enzymatic reaction. An in vitro multi-enzymatic system and a heterologous co-expression system will be developed, respectively, for yellow MP biosynthesis. This research will elucidate the biosynthetic pathway of yellow MP, and establish theory foundation to develop new yellow MP producing Monascus strains. Furthermore, this will provide reference for studying the biosynthetic pathway of analogous metabolites in other fungi.