AI Article Synopsis
- The study examines the ability of a Rhodococcus erythropolis mutant strain, RG9, to biotransform norandrostenedione using a cytochrome P450 BM3 mutant enzyme (M02).
- The purified enzyme demonstrated over 95% regioselectivity in hydroxylating the steroid, producing 16-β-OH norandrostenedione, which was confirmed through NMR analysis.
- Whole cells of RG9 achieved the conversion of norandrostenedione to the hydroxylated product without any activity from the strain alone, highlighting the potential of R. erythropolis for effective steroid bioconversions using this enzyme.
Article 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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| http://dx.doi.org/10.1007/s00253-014-6281-7 | DOI Listing |
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