Collaborative Research: Genetic Analysis and Metabolic Engineering of Solventogenic Clostridia

合作研究：产溶剂梭菌的遗传分析和代谢工程

• 批准号: 9604562
• 负责人: George Bennett
• 金额: --
• 依托单位: William Marsh Rice University
• 依托单位国家: 美国
• 项目类别: Continuing grant
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-15 至 2000-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9604562&HistoricalAwards=false
• 关键词: Collaborative; Research; Genetic; Analysis; Metabolic

9604562 Bennett This proposal is on research to genetically alter the metabolism of anaerobic solventogenic clostridia (Clostridium acetobutylicum and related species) to determine their potential applications for producing solvents (butanol and acetone) and other oxychemicals. This requires further understanding of the applied molecular genetics and associated methods needed to redirect the cellular metabolism. This research is aiming to (1) establish the techniques needed for the generation of stable overproducing strains (such strains are necessary for batch or continuous processes without using any antibiotics to maintain genetic stability), (2) develop a more detailed molecular understanding of the regulation of a cluster of genes known as the sol locus, (3) develop the means which would allow one to express clostridia genes in other hosts or heterologous genes in clostridia in order to form products not normally produced by the respective cells, and(4) develop additional tools for flexible carbon-flux control. Specifically, this research will develop methods needed for: desirable gene expression (in terms of strength and timing), single or multiple chromosomal gene inactivations in order to inactivate or eliminate undesirable pathways or traits, and stable chromosomal integration (of one or more copies) of genes in order to increase the protein activities (and presumably carbon fluxes) of desirable pathways. ***

9604562 Bennett 该提案旨在研究从基因角度改变厌氧产溶剂梭菌（丙酮丁醇梭菌及相关物种）的代谢，以确定其在生产溶剂（丁醇和丙酮）和其他含氧化学品方面的潜在应用。 这需要进一步了解应用分子遗传学和重定向细胞代谢所需的相关方法。 这项研究的目的是（1）建立产生稳定过量生产菌株所需的技术（此类菌株是批量或连续过程所必需的，无需使用任何抗生素来维持遗传稳定性），（2）对调节称为 sol 位点的一组基因，(3) 开发允许在其他宿主中表达梭菌基因或在梭菌中表达异源基因的方法，以形成各个细胞通常不产生的产物，以及（4 ）开发用于灵活碳通量控制的附加工具。 具体来说，这项研究将开发以下所需的方法：所需的基因表达（在强度和时间方面）、单个或多个染色体基因失活以失活或消除不需要的途径或性状，以及稳定的染色体整合（一个或多个拷贝）基因以增加所需途径的蛋白质活性（以及可能的碳通量）。 ***