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Biofuels are produced from living organisms by metabolic products. We are in the process of developing a new green technology that can produce a biodiesel from gram-positive bacteria, Bacillus subtilis 168. Naturally, most bacteria produce fatty acids as a cell envelope
precursor, and multiple steps are needed to synthesize fatty acids. Among them, acetyl-CoA carboxylase (encoded by accA, accB, accC) is a multi-subunit key enzyme for the biosynthesis of fatty acids. The main function of an acetyl-CoA carboxylase is catalyzing the biotindependent
carboxylation of acetyl-CoA to provide malonyl-CoA. This process is the most essential step in the biosynthesis of long-chain fatty acids. Therefore, overexpression of the acetyl-CoA carboxylase subunits increases the productivity of fatty acids. We have engineered
a competent producer of fatty acids via introducing three different genes into the Bacillus subtilis genome. In this study, we constructed two recombinant plasmids, pET22bA and pET22bBC. These two plasmids were transformed into Bacillus subtilis 168 and tried to express the target genes consecutively. The compounds produced by two distinct recombinant strains have been analyzing by GC. This study implies that these genes encode the key enzyme for fatty acid
biosynthesis in Bacillus subtilis, and ultimately indicates that a large amount of biodiesel can be produced by metabolic engineering.