酿酒酵母工程化外源途径的转录精细调控基础

项目名称： 酿酒酵母工程化外源途径的转录精细调控基础

项目编号： No.21476026

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 有机化学

项目作者： 周晓宏

作者单位： 北京理工大学

项目金额： 85万元

中文摘要： 针对外源代谢途径在酿酒酵母工程菌中转录过程难以精细调控的问题，本项目运用合成生物学方法开展酿酒酵母精细调控元件的发掘与功能应用研究。首先研究酿酒酵母启动子的结构与功能，发掘新的启动子，获得新的酿酒酵母启动子文库；其次对启动子序列进行分析，筛选出对启动子功能具有重要影响作用的转录因子结合位点，阐明转录因子结合位点对启动子特性影响的分子机制；然后以此为基础构建出启动子强度可控的启动子元件，将启动子强度的离散性转变为连续性；最后，在酿酒酵母中将精细调控启动子元件与课题组前期已构建好的初生代谢木糖醇合成途径和次生代谢β-香树脂醇合成途径进行组装，以进行启动子调控元件的功能验证。项目的实施可望通过启动子调控元件精确调控酿酒酵母中外源代谢途径或者分支代谢途径的代谢流，提高代谢转化效率，为代谢工程的途径调控研究提供思路。

中文关键词： 转录；调控；酿酒酵母；木糖醇；香树脂醇

英文摘要： For the difficulty in fine transcription regulation of heterologous metabolic pathway in Saccharomyces cerevisiae, the promoter regulatory elements and their functional appilication will be investigated by using synthetic biology in this research. First, the new promoters will be discovered through studying the structures and functions of the present promoters in S. cerevisiae and a new promoter library will be constructed. Second, after the analysis of the sequences of promoters, the transcription factor binding sites (TFBSs) which are important to the function of promoters will be screened and the molecular mechanisms of the effect of TFBSs on promoters will be clarified. Then, by using these TFBSs, a series of fine tunable promoters will be constructed, in which their strengths will not be discrete but continuous. At last, a typical primary synthetic pathway of xylitol and a typical secondary synthetic pathway of β-amyrin previously reconstructed in our group will be chosen to verify the function of the promoter regulatory elements in S. cerevisiae. In this research, the flux not only in heterologous pathway but also branched pathway will be balanced through the promoter regulatory elements and the biosynthetic efficiency will be expectantly improved, which will also provide ideas for the regulation of engineered metabolic pathway.

英文关键词： transcription;regulation;Saccharomyces cerevisiae;xylitol;amyrin