Objective Regio- and stereoselective hydroxylation of lithocholic acid (LCA) using CYP107D1 (OleP), a cytochrome P450 monooxygenase from the oleandomycin synthesis pathway of Streptomyces antibioticus . Results Co-expression of CYP107D1 from S. antibioticus and the reductase/ferredoxin system PdR/PdX from Pseudomonas putida was performed in Escherichia coli whole cells. In vivo hydroxylation of LCA exclusively yielded the 6β-OH product murideoxycholic acid (MDCA). In resting cells, 19.5% of LCA was converted to MDCA within 24 h, resulting in a space time yield of 0.04 mmol L −1 h −1 . NMR spectroscopy confirmed the identity of MDCA as the sole product. Conclusions The multifunctional P450 monooxygenase CYP107D1 (OleP) can hydroxylate LCA, forming MDCA as the only product.

中文翻译：

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多功能细菌 P450 单加氧酶 CYP107D1 (OleP) 的高选择性胆汁酸羟基化

目的 使用 CYP107D1 (OleP)，一种来自抗生素链霉菌的夹竹桃霉素合成途径的细胞色素 P450 单加氧酶，对石胆酸 (LCA) 进行区域和立体选择性羟基化。结果 来自抗生素链球菌的 CYP107D1 和来自恶臭假单胞菌的还原酶/铁氧还蛋白系统 PdR/PdX 的共表达在大肠杆菌全细胞中进行。LCA 的体内羟基化仅产生 6β-OH 产物鼠氧胆酸 (MDCA)。在静息细胞中，19.5% 的 LCA 在 24 小时内转化为 MDCA，导致时空产率为 0.04 mmol L -1 h -1 。核磁共振光谱证实了 MDCA 是唯一的产物。结论多功能P450单加氧酶CYP107D1（OleP）可以羟基化LCA，形成MDCA作为唯一产物。