Biosynthesis of a steroid metabolite by an engineered Rhodococcus erythropolis strain expressing a mutant cytochrome P450 BM3 enzyme
Biosynthesis of a steroid metabolite by an engineered Rhodococcus erythropolis strain expressing a mutant cytochrome P450 BM3 enzyme
复制标题
DOI：
10.1007/s00253-014-6281-7
复制
发表时间：
2015-06
影响因子：
5
通讯作者：
H. Venkataraman;E. T. Poele;Kamila Z. Rosłoniec;N. Vermeulen;J. Commandeur;R. Geize;L. Dijkhuizen
中科院分区：
文献类型：
--
作者：
H. Venkataraman;E. T. Poele;Kamila Z. Rosłoniec;N. Vermeulen;J. Commandeur;R. Geize;L. Dijkhuizen
在本研究中，使用ofRhodococcus erythropolis突变株RG 9表达的细胞色素P450 BM 3突变M02酶已被评估为全细胞生物转化的17-酮类固醇，去甲雄烯二酮，作为一个模型底物。纯化的P450 BM 3突变体M02酶以> 95%的区域选择性羟基化类固醇以形成16-β-OH去甲雄烯二酮，如通过NMR分析所证实的。R.表达P450 BM 3 M02酶的erythropolisRG 9也将去甲雄烯二酮转化为16-β-羟基化产物，导致形成约0.35 g/L。菌株RG 9本身的全细胞不转化去甲雄烯二酮，表明代谢物形成是P450 BM 3 M02酶介导的。研究表明，R.红细胞生成菌是一种新的和令人感兴趣的宿主，用于异源表达能够进行类固醇生物转化的高选择性和活性的P450 BM 3 M02酶变体。
In the present study, the use ofRhodococcus erythropolismutant 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 ofR. erythropolisRG9 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 thatR. erythropolisis a novel and interesting host for the heterologous expression of highly selective and active P450 BM3 M02 enzyme variants able to perform steroid bioconversions.