CYP109E1 from Bacillus megaterium Acts as a 24‐ and 25‐Hydroxylase for Cholesterol

Abstract

In this study, the ability of CYP109E1 from Bacillus megaterium DSM319 to metabolize cholesterol was investigated. This steroid was identified as a new substrate to be converted by CYP109E1 with adrenodoxin and adrenodoxin reductase as redox partners in vitro. The biotransformation was successfully reproduced in vivo by using Bacillus megaterium cells that overexpressed CYP109E1. To enhance the production of cholesterol derivatives, an Escherichia coli based whole‐cell system that harbored CYP109E1 was established. This novel system showed a 3.3‐fold higher activity than that of the B. megaterium system, yielding about 45 mg L−1 of these products. Finally, the reaction products were isolated and identified to be the highly important cholesterol derivatives 24(S)‐ and 25‐hydroxycholesterol.

Plant growth-promoting activities and genomic analysis of the stress-resistant Bacillus megaterium STB1, a bacterium of agricultural and biotechnological interest

“…In addition, the genome of strain STB1 contains several cytochrome p450 encoding genes ( Table S7 ), including a cytochrome P450/NADPH--P450 reductase homolog (C0569_02245) (98.1 % identity to B. megaterium ATCC 14581 CYP102A1) involved in fatty acid hydroxylation [43]; a CYP109E1 homolog (C0569_04895) (98.5 % identity to B. megaterium DSM 319 CYP109E1) involved in the hydroxylation of steroids, vitamin D3 (cholecalciferol), cholesterol, statins and terpenes [[44], [45], [46],47]; a CYP106A1 homolog (C0569_20645) (95.4 % identity to B. megaterium DSM 319 CYP106A1) involved in the hydroxylation of the pentacyclic triterpene 11-keto-β-boswellic acid and several steroids [48,49]; a CYP109A2 homolog (C0569_21565) (97.3 % identity to B. megaterium DSM 319 CYP109A2) involved in vitamin D3 hydroxylation [50]; and a unknown cytochrome P450 (C0569_21365) with 96 % identity to a B. megaterium DSM 319 WP_080514752.1 cytochrome P450.…”

Evolution of Cytochrome P450 Enzymes and Their Redox Partners in Archaea

“…CYP109D1 of S. cellulosum was also shown to have highly regioselective hydroxylation of norisoprenoids, alpha- and beta-ionone [ 56 ]. CYP109E1 from Bacillus megaterium catalyzes a cholesterol and vitamin D2 two-step hydroxylation at positions C24 and C25 [ 57 , 58 ].…”

Rational development of mycobacteria cell factory for advancing the steroid biomanufacturing

“…3 ). For example, 25-hydroxycholesterol possesses a significant antiviral and counteract inflammation activity (Diczfalusy, 2013 ; Tricarico et al 2019 ), and could be produced from cholesterol by CYP109E1 from Bacillus megaterium (Putkaradze et al 2019 ). CYP11A1 can biosynthesize pregnenolone by diametrically catalyzing the side-chain hydroxylation and cleavage of cholesterol or phytosterols, which avoids multiple rounds of step-by-step degradation (Zhang et al 2019a ).…”