Steroids hydroxylation catalyzed by the monooxygenase mutantfromBM3.
10.1016/j.apsb.2017.04.006
- Author:
Xing LIU
1
;
Jian-Qiang KONG
1
Author Information
1. State Key Laboratory of Bioactive Substance and Function of Natural Medicines & Ministry of Health Key Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.
- Publication Type:Journal Article
- Keywords:
139-3;
1α-OH-androstenedione;
BM3;
Biocatalyst;
Cytochrome P450;
Steroids hydroxylation
- From:
Acta Pharmaceutica Sinica B
2017;7(4):510-516
- CountryChina
- Language:English
-
Abstract:
The search of new substrates with pharmaceutical and industrial potential for biocatalysts including cytochrome P450 enzymes is always challenging. Cytochrome P450 BM3 mutant, a versatile biocatalyst, exhibited hydroxylation activities towards fatty acids and alkanes. However, there were limited reports about its hydroxylation activity towards steroids. Herein, an-based whole-cell extract containing the recombinant 139-3 protein was used as the biocatalyst to screen 13 steroids. Results revealed that 139-3 was able to specifically hydroxylate androstenedione () at 1-position, generating a hydroxylated steroid 1-OH-androstenedione (). To investigate whether C-1hydroxylation catalyzed by BM3 mutantcould be industrially used, an optimization of catalyzing conditions was performed. Accordingly, the BM3 mutant 139-3 enzyme was observed to display maximum activity at 37 °C, under pH 7.0 for 4 h, with 37% transformation rate. Moreover, fourvariants were generated by random mutagenesis with the aim of improving its activity and expanding substrate scope. Surprisingly, these mutants, sharing a common mutated site R379S, lost their activities towards androstenedione (). These data clearly indicated that arginine residue located at site 379 played key role in the hydroxylation activities of 139-3. Overall, these new findings broadened the substrate scope of 139-3 enzyme, thereby expanding its potential applications as a biocatalyst on steroids hydroxylation in pharmaceutical industry.
