Journal article 311 views 86 downloads
Structure–Function Analysis of the Steroid-Hydroxylating Cytochrome P450 109 (CYP109) Enzyme Family
,
Thembeka Khumalo,
David R. Nelson ,
David Lamb ,
Khajamohiddin Syed
International Journal of Molecular Sciences, Volume: 26, Issue: 13, Start page: 6219
Swansea University Author:
David Lamb
-
PDF | Version of Record
© 2025 by the authors. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license.
Download (5.23MB)
DOI (Published version): 10.3390/ijms26136219
Abstract
Steroids are found in bacteria and eukaryotes, and genes potentially encoding steroid metabolic enzymes have also been identified in giant viruses. For decades, hydroxylated steroids have been utilized in medicine to treat various human diseases. The hydroxylation of steroids can be achieved using m...
| Published in: | International Journal of Molecular Sciences |
|---|---|
| ISSN: | 1422-0067 |
| Published: |
MDPI
2025
|
| Online Access: |
Check full text
|
| URI: | https://cronfa.swan.ac.uk/Record/cronfa69940 |
| Abstract: |
Steroids are found in bacteria and eukaryotes, and genes potentially encoding steroid metabolic enzymes have also been identified in giant viruses. For decades, hydroxylated steroids have been utilized in medicine to treat various human diseases. The hydroxylation of steroids can be achieved using microbial enzymes, especially cytochrome P450 monooxygenases (CYPs/P450s) and is well documented. Understanding the structural determinants that govern the regio- and stereoselectivity of steroid hydroxylation by P450s is essential in order to fully exploit their potential. Herein, we present a comprehensive analysis of the steroid-hydroxylating CYP109 family across the domains of life and delineate the structural determinants that govern steroid hydroxylation. Data mining, annotation, and phylogenetic analysis revealed that CYP109 family members are highly populated in bacteria, and indeed, these members passed from bacteria to archaea by horizontal gene transfer, leading to the evolution of P450s in archaea. Analysis of twelve CYP109 crystal structures revealed large, flexible, and dynamic active site cavities that can accommodate multiple ligands. The correct positioning and orientation of the steroid in the active site cavity and the nature of the C17 substituent on the steroid molecule influence catalysis. In an analogous fashion to the CYP107 family, the amino acid residues within the CYP109 binding pocket involve hydrophilic and hydrophobic interactions, influencing substrate orientations and anchoring and determining the site of hydroxylation and catalytic activity. A handful of amino acids, such as Val84, Val292, and Ser387 in CYP109B4, have been found to play a role in determining the catalytic regiospecificity, and a single amino acid, such as Arg74 in CYP109A2, has been found to be essential for the enzymatic activity. This work serves as a reference for the precise understanding of CYP109 structure–function relationships and for P450 enzymes in general. The findings will guide the genetic engineering of CYP109 enzymes to produce valuable steroid molecules of medicinal and biotechnological importance. |
|---|---|
| Keywords: |
cytochrome P450; CYP109; steroid; oxidation reactions; active site; crystal structure |
| College: |
Faculty of Medicine, Health and Life Sciences |
| Funders: |
National Research Foundation (NRF), South Africa Grant: PMDS230527110616 Grant: MND210504599108; National Research Foundation (NRF), South Africa Grant: RA22102865602 |
| Issue: |
13 |
| Start Page: |
6219 |