Highly selective bile acid hydroxylation by the multifunctional bacterial P450 monooxygenase CYP107D1 (OleP)

Sascha Grobe,Henrike Brundiek,Melinda Fekete,Agata Wszołek,Uwe T Bornscheuer

doi:10.1007/s10529-020-02813-4

Abstract

ObjectiveRegio- and stereoselective hydroxylation of lithocholic acid (LCA) using CYP107D1 (OleP), a cytochrome P450 monooxygenase from the oleandomycin synthesis pathway of Streptomyces antibioticus.ResultsCo-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.ConclusionsThe multifunctional P450 monooxygenase CYP107D1 (OleP) can hydroxylate LCA, forming MDCA as the only product.

Highlights

Bile acids and their derivatives are steroidal compounds and play an important biological role in digestion by the solubilization of vitamins and fatty acids (Mikov et al 2006).Besides their biological function, bile acids have become an interesting compound class for pharmaceutical applications (Hofmann and Hagey 2008)
We found that CYP107D1 stereo- and regioselectively hydroxylates lithocholic acid (LCA) and deoxycholic acid (DCA) at the 6b-position
Whole cells of E. coli C43 (DE3) co-expressing OleP (CYP107D1) and PdR/PdX converted LCA (2 mg mL-1, 5.31 mM) to a single product according to HPLC analysis

Summary

Introduction

Bile acids and their derivatives are steroidal compounds and play an important biological role in digestion by the solubilization of vitamins and fatty acids (Mikov et al 2006).Besides their biological function, bile acids have become an interesting compound class for pharmaceutical applications (Hofmann and Hagey 2008). CDCA (Roda et al 1982), MDCA (Cohen et al 1990, 1991) and UDCA (Leuschner et al 1985) are valuable pharmaceutical agents, because they can be used to solubilize gallstones These bile acid derivatives are applied in treatments of gallstones and liver disease (Chiang 2017). Up to today the synthesis of valuable bile acids and their derivatives is difficult and the efficiency of the processes is low as reviewed by Tonin and Arends (Tonin and Arends 2018). This is caused by multistep chemical or chemoenzymatic routes with toxic intermediates, protection and deprotection steps to control regio- and stereoselectivity of hydroxylations, and modifications of the steroid nucleus (Eggert et al 2014)

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Results

Conclusion