ME: Redirecting Cellular Metabolism for the Biodegradation of Mixtures of Chlorinated Solvents

ME：重定向细胞代谢以实现氯化溶剂混合物的生物降解

• 批准号: 0004248
• 负责人: Kenneth Reardon
• 金额: $ 36.3万
• 依托单位: Colorado State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-15 至 2003-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0004248&HistoricalAwards=false
• 关键词: ME; Redirecting; Cellular; Metabolism; Biodegradation

The objective of this project is to create a new paradigm in bioremediation of chlorinated solvents by metabolically engineering microorganisms for the rapid, long-term, aerobic biodegradation of perchloroethylene (PCE), trichloroethylene (TCE), and mixtures of these and other solvents. No organisms are known to have these abilities, and development of such a strain would provide a new, cost-effective option for solving some of the world's most pervasive soil and groundwater pollution problems. This research has four specific objectives: (1) use DNA shuffling to create an optimized monooxygenase that degrades PCE, TCE, and mixtures of chlorinated solvents; (2) clone this monooxygenase and epoxide hydrolase into two host strains; (3) evaluate the effects of this genetic engineering on host cell physiology using 2-dimensional protein electrophoresis; and (4) evaluate the fate of modified cells and introduced genes in soil and bioreactor settings.

本项目的目标是通过代谢工程微生物对四氯乙烯（PCE）、三氯乙烯（TCE）以及这些溶剂和其他溶剂的混合物进行快速、长期、好氧生物降解，从而在氯化溶剂的生物修复方面创建一个新的范例。 目前还没有已知的生物具有这些能力，开发这样一种菌株将为解决世界上一些最普遍的土壤和地下水污染问题提供一种新的、具有成本效益的选择。 本研究有四个具体的目的：（1）利用DNA改组技术优化单加氧酶基因，以降解PCE、TCE和氯化溶剂的混合物;（2）将该单加氧酶和环氧化物水解酶基因克隆到两种宿主菌中;（3）利用蛋白质双向电泳技术评价该基因工程对宿主细胞生理的影响;和（4）评估修饰细胞和引入的基因在土壤和生物反应器环境中的命运。