Biosynthesis of a steroid metabolite by an engineered Rhodococcus erythropolis strain expressing a mutant cytochrome P450 BM3 enzyme

Harini Venkataraman, Evelien M. te Poele, Kamila Z. Rosłoniec, Nico Vermeulen, Jan N.M. Commandeur, Robert van der Geize, Lubbert Dijkhuizen*

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

In the present study, the use of Rhodococcus erythropolis mutant strain RG9 expressing the cytochrome P450 BM3 mutant M02 enzyme has been evaluated for whole-cell biotransformation of a 17-ketosteroid, norandrostenedione, as a model substrate. Purified P450 BM3 mutant M02 enzyme hydroxylated the steroid with >95 % regioselectivity to form 16-β-OH norandrostenedione, as confirmed by NMR analysis. Whole cells of R. erythropolis RG9 expressing P450 BM3 M02 enzyme also converted norandrostenedione into the 16-β-hydroxylated product, resulting in the formation of about 0.35 g/L. Whole cells of strain RG9 itself did not convert norandrostenedione, indicating that metabolite formation is P450 BM3 M02 enzyme mediated. This study shows that R. erythropolis is a novel and interesting host for the heterologous expression of highly selective and active P450 BM3 M02 enzyme variants able to perform steroid bioconversions.

Original languageEnglish
Pages (from-to)4713-4721
Number of pages9
JournalApplied Microbiology and Biotechnology
Volume99
Issue number11
DOIs
Publication statusPublished - 18 Jun 2015

Keywords

  • Cytochrome P450 BM3 M02 enzyme
  • Norandrostenedione
  • Rhodococcus erythropolis
  • Steroids
  • Whole-cell bioconversion

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