Collaborative Research: Metabolite-responsive regulators for polyketide pathway engineering

合作研究：聚酮化合物途径工程的代谢物响应调节剂

• 批准号: 1159398
• 负责人: Patrick Cirino
• 金额: $ 29.91万
• 依托单位: University of Houston
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1159398&HistoricalAwards=false
• 关键词: Collaborative; Research; Metabolite; responsive; regulators

1159398/1159759 Cirino/TangPolyketides are an important class of bioactive natural products. The complexity of these compounds demands that their synthesis be conducted by dedicated enzymes with exquisite catalytic properties. Directed evolution of natural product pathway enzymes is limited by the lack of general high-throughput screens for the complex target compounds, as well as the genetic intractability of most organisms expressing these pathways. The proposed research addresses this technology gap through the engineering of synthetic genetic networks that can respond to a desired polyketide compound. Many bacterial regulatory proteins naturally control gene expression in response to a specific "effector" molecule. We will screen mutant libraries of regulatory proteins to isolate variants that activate expression of a reporter gene in response to polyketide compounds of interest. Then, using E. coli outfitted with the synthetic regulatory circuit, the aromatic polyketide biosynthetic pathway will be engineered towards enhanced production of bioactive metabolites and biosynthesis of new, "unnatural" compounds.This project is cofunded by the Biotehchnology, Biochemical and Biomass Engineering Program, Chemical, Bioengineerng, Environmental and Transport Systems Division, Engineering Directorate, and the Chemistry of Life Processes Program in the Chemistry Division of MPS.

聚酮类化合物是一类重要的生物活性天然产物。这些化合物的复杂性要求它们的合成必须由具有精细催化性能的专用酶来进行。天然产物途径酶的定向进化受到复杂目标化合物缺乏一般高通量筛选以及大多数表达这些途径的生物体的遗传难解性的限制。提出的研究解决了这一技术差距，通过工程合成的遗传网络，可以响应所需的聚酮化合物。许多细菌调节蛋白自然控制基因表达，以响应特定的“效应”分子。我们将筛选调节蛋白的突变文库，以分离对感兴趣的聚酮化合物激活报告基因表达的变异。然后，使用配备了合成调控电路的大肠杆菌，芳香聚酮生物合成途径将被设计成增强生物活性代谢物的生产和新的“非自然”化合物的生物合成。该项目由生物技术、生化和生物质工程项目、化学、生物工程、环境和运输系统部门、工程局和MPS化学部门的生命过程化学项目共同资助。