Premium
Structural characterization of CYP 260A1 from Sorangium cellulosum to investigate the 1α‐hydroxylation of a mineralocorticoid
Author(s) -
Khatri Yogan,
Carius Yvonne,
Ringle Michael,
Lancaster C. Roy D.,
Bernhardt Rita
Publication year - 2016
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1002/1873-3468.12479
Subject(s) - adrenodoxin , hydroxylation , chemistry , asparagine , stereochemistry , steroid , cytochrome p450 , mutant , crystal structure , biochemistry , enzyme , crystallography , hormone , gene
In this study, we report the crystal structure of the cytochrome P450 CYP 260A1 (PDB 5LIV ) from the myxobacterium Sorangium cellulosum So ce56. In addition, we investigated the hydroxylation of 11‐deoxycorticosterone by CYP 260A1 by reconstituting the enzyme with the surrogate redox partners adrenodoxin and adrenodoxin reductase. The major product of this steroid conversion was identified as 1α‐hydroxy‐11‐deoxycorticosterone, a novel Δ4 C‐21 steroidal derivative. Furthermore, we docked the substrate into the crystal structure and replaced Ser326, the residue responsible for substrate orientation, with asparagine and observed that the mutant S326N displayed higher activity and selectivity for the formation of 1α‐hydroxy‐11‐deoxycorticosterone compared to the wild‐type CYP 260A1. Thus, our findings highlight the usefulness of the obtained crystal structure of CYP 260A1 in identifying biotechnologically more efficient reactions.