Collaborative Research: Metabolite-responsive regulators for polyketide pathway engineering

Cirino/TangPolyketides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由具有精致催化特性的专用酶进行。天然产物途径酶的定向进化受到缺乏对复杂目标化合物的一般高通量筛选以及表达这些途径的大多数生物体的遗传难治性的限制。拟议的研究通过设计合成遗传网络来解决这一技术差距，该网络可以对所需的聚酮化合物做出反应。许多细菌调节蛋白响应于特定的“效应物”分子而天然地控制基因表达。 我们将筛选调控蛋白的突变体文库，以分离出响应于感兴趣的聚酮化合物而激活报告基因表达的变体。然后，利用E.该项目由生物技术、生物化学和生物质工程项目、化学、生物工程、环境和运输系统部、工程理事会、和MPS化学部的生命过程化学计划。

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.