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.

中文翻译：

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麝香大环内酯从脂肪酸的酶促级联生物合成反应

麝香大环内酯是高价值香水中使用的一组重要化合物。探索了包括细胞色素 P450 羟化酶和脂肪酶在内的酶促级联反应来生物合成麝香大环内酯。首先，脂肪酸被P450羟化酶羟基化，产生相应的ω-羟基脂肪酸。然后ω-羟基脂肪酸被脂肪酶内酯化。ω-羟基脂肪酸难以通过传统的化学方法合成，而这些化合物的产生是整个反应过程中的关键制约因素。为了获得足够的大环内酯前体，探索了 ω-羟基脂肪酸的有效生产。来自巨大芽孢杆菌的 P450 BM3 突变体用作末端羟化酶。为了提高ω-羟基脂肪酸的产量，优化了辅酶再生系统和辅助有机溶剂。在双相系统下使用 P450 BM3 突变体的转化率高达 42% 向 ω-羟基十五烷酸和 98% 向 ω-羟基棕榈酸。结果表明，麝香大环内酯、exaltolide和silvanone supra，可以在羟基化-内酯化级联反应中合成。最后，从 242mg 十五烷酸得到 81mg exaltolide，从 256mg 棕榈酸得到 199mg silvanone supra。