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This research illustrates the development of microbial production of fatty acid, which can be transformed into the bio-diesel for alternative energy. We developed a new green technology that can produce a fatty acid with high productivity from both strains, Bacillus subtilis (gram-positive) and Escherichia coli (gram-negative). To produce a fatty acid in bacteria, it is essential to generate a three-carbon precursor, malonyl-CoA, which is derived from acetyl-CoA by the action of acyl-CoA carboxylase (accA, accB, accC). The malonyl-CoA is transferred to the acyl carrier protein by malonyl-CoA:ACP acyltransferase (FabD). This process is the most essential step in the biosynthesis of long–-chain fatty acids. Therefore, overexpression of accA, accB and accC genes, encoding acetyl-CoA carboxylase subunits enzymes, is expected to increase the productivity of fatty acids. The single, double and triple organized artificial operons of accA, accB, accC and fabD were designed and constructed in a vector. By introducing four distinct genes obtained from Bacillus subtilis into E. coli, we have engineered E. coli strain for high productivity of fatty acids.