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

基本信息

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

项目摘要

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.

1159398/1159759 Cirino/Tang 聚酮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由具有精细催化特性的专用酶进行。由于缺乏对复杂目标化合物的通用高通量筛选，以及表达这些途径的大多数生物体的遗传难处理性，天然产物途径酶的定向进化受到限制。拟议的研究通过合成遗传网络的工程来解决这一技术差距，该网络可以响应所需的聚酮化合物。许多细菌调节蛋白天然地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体文库，以分离出响应目标聚酮化合物而激活报告基因表达的变体。然后，利用配备有合成调节回路的大肠杆菌，将设计芳香族聚酮化合物生物合成途径，以提高生物活性代谢物的产量和新的“非天然”化合物的生物合成。该项目由生物技术、生物化学和生物质工程计划共同资助、化学、生物工程、环境和运输系统部、工程局以及公安部化学部的生命过程化学项目。