Collaborative Research: Genetic Analysis and Metabolic Engineering of Solventogenic Clostridia

9632217 Papoutsakis这项建议是为了从基因上改变厌氧产溶血性梭菌(乙酰丁基梭菌和相关物种)的新陈代谢，以确定它们在生产溶剂(丁醇和丙酮)和其他氧化剂方面的潜在应用。这需要进一步了解应用分子遗传学以及改变细胞代谢方向所需的相关方法。这项研究的目的是(1)建立产生稳定的高产菌株所需的技术(这种菌株是批量或连续生产所必需的，而不需要使用任何抗生素来维持遗传稳定性)；(2)对称为SO/基因座的一组基因的调节有更详细的分子理解；(3)开发允许一个人在其他宿主中表达梭状芽胞杆菌基因或在梭状芽胞杆菌中表达异源基因的方法，以便形成不是由各自的细胞正常产生的产品；以及(4)开发更多的灵活碳通量控制工具。具体地说，这项研究将开发以下方法：理想的基因表达(在强度和时间方面)，单个或多个染色体基因的失活以灭活或消除不希望的途径或特征，以及稳定的基因染色体整合(一个或多个副本)，以便增加理想途径的蛋白质活性(可能还有碳通量)。***

9632217 Papoutsakis 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 so/ 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 need 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. ***