Enzymatic cascade biosynthesis reaction of musky macrolactones from fatty acids

Abstract

Musky macrolactones are an important group of compounds used in high-valued perfumery. An enzymatic cascade reaction including cytochrome P450 hydroxylase and lipase was explored to biosynthesize musky macrolactones. Firstly, fatty acids were hydroxylated by P450 hydroxylase to produce the corresponding ω-hydroxy fatty acids. Then ω-hydroxy fatty acids were lactonized by lipase. ω-Hydroxy fatty acids can difficultly be synthesized by traditional chemical methods, and the production of these compounds were key constraint factors during the whole reaction. To obtain enough precursors of macrolactones, an efficient production of ω-hydroxy fatty acids was explored. A mutant of P450 BM3 from Bacillus megaterium was used as terminal hydroxylases. To improve the yield of ω-hydroxy fatty acids, the coenzyme regeneration system and auxiliary organic solvent were optimized. The conversion using the P450 BM3 mutant under the biphase system was up to 42% towards ω-hydroxy pentadecanoic acid and 98% towards ω-hydroxy palmitic acid. The results reveal that the musky macrolactones, exaltolide and silvanone supra, could be synthesized in the hydroxylation-lactonization cascade reaction. Finally, 81 mg of exaltolide was obtained from 242 mg pentadecanoic acid, and 199 mg of silvanone supra from 256 mg palmitic acid.