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

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.

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Acknowledgments

This work was financially supported by the Netherlands Organization for Scientific Research (NWO) within the IBOS (Integration of Biosynthesis and Organic Synthesis) program.

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Correspondence to Lubbert Dijkhuizen.

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Harini Venkataraman and Evelien M. te Poele contributed equally.

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Venkataraman, H., te Poele, E.M., Rosłoniec, K.Z. et al. Biosynthesis of a steroid metabolite by an engineered Rhodococcus erythropolis strain expressing a mutant cytochrome P450 BM3 enzyme. Appl Microbiol Biotechnol 99, 4713–4721 (2015). https://doi.org/10.1007/s00253-014-6281-7

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Keywords

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