Efficient synthesis of malonyl-CoA by an acyl-CoA synthetase from Streptomyces sp.

Malonyl-CoA is a precursor of fatty acids, polyketides, and bio-based chemicals with potential applications in medicine, antibiotics, and fuels. However, its low intracellular concentration and high cost have led to difficulties in research and production. To develop an efficient method for producing malonyl-CoA, we screened the acyl-CoA synthetase (ACS) gene from Streptomyces sp. using sequence-structure alignment. This protein contains conserved sequences and active sites for malonyl-CoA synthetases. The purified recombinant enzyme ACS was heterologously expressed in Escherichia coli BL21 and characterised. The results showed that it converted the substrates malonate and CoA into malonyl-CoA. Under the optimal conditions, the specific activity of the purified ACS was 32.3 U·mg−1 and the conversion rate reached 98.8%. In addition, when the cell-free extracts were used as catalysts, the highest yield of malonyl-CoA was obtained after 4 h, yielding 24.2 g·L−1 with a conversion rate of 90.3%. After the product was purified and vacuum freeze-dried, a solid powder of malonyl-CoA was obtained. This study characterised and identified a new ACS and optimised the reaction conditions to efficiently synthesise pure malonyl-CoA in vitro in high yield using enzyme-mediated methods.