Development of efficient production platform of novel antimicrobial agents by malonyl-CoA overproduction and studying the antimicrobial efficacy
Shah, Karan Prakash
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Genetically tractable microorganisms, such as Escherichia coli , serve as ideal platform for recombinant protein expression and heterologous biosynthesis of a variety of small molecules with varying health benefits. However, in many cases protein activities and biosynthetic efficiency are greatly limited by precursor and co-factor availabilities in the recombinant host. Malonyl-CoA is a critical metabolite for the biosynthesis of numerous fine chemicals of industrial significance including fatty acids, flavonoids and polyketides to name a few. In the present work we alter intracellular levels of Malonyl-CoA by deleting and/or over expressing fatty acid synthesis pathways and regulators. These selected targets should lead to inhibition of Malonyl-CoA consumption in the biosynthetic pathway. To assess the impact of the genetic modifications, the plant-derived pathway for biosynthesis of flavanones, the primary flavonoid, was introduced on co-replicable vectors to then examine flavanone production by shake flask fermentation. One important aspect in developing efficient recombinant producers of natural products such as flavonoids is their toxic effects on the recombinant host. Especially in the case of flavonoids, it is known that they have antibiotic properties and are used by plants to fight microbial infections. We are synthesizing various non-natural flavonoid compounds with substituent halogens at different positions in the structure in order to create compounds with novel properties. Therefore we are also interested in testing the anti-microbial properties by determining Minimum Inhibitory Concentration (MIC) values of the various novel flavonoid compounds synthesized Based on the results obtained, we aim to synthesize compounds with high antibiotic activity, either by themselves or in combination with efflux pump inhibitors similar to that of potent antibiotics.