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. ***
