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Engineering of redox partners and cofactor NADPH supply of CYP68JX for efficient steroid two-step ordered selective hydroxylation activity.
- Source :
-
Journal of Steroid Biochemistry & Molecular Biology . Apr2024, Vol. 238, pN.PAG-N.PAG. 1p. - Publication Year :
- 2024
-
Abstract
- CYP68JX, a P450 hydroxylase, derived from Colletotrichum lini ST-1 is capable of biotransforming dehydroepiandrosterone (DHEA) to 3β,7α,15α-trihydroxy-5-androstene-17-one (7α,15α-diOH-DHEA). Redox partners and cofactor supply are important factors affecting the catalytic activity of CYP68JX. In this study, the heterologous expression of CYP68JX in Saccharomyces cerevisiae BY4741 was realized resulting in a 17.1% target product yield. In order to increase the catalytic efficiency of CYP68JX in S. cerevisiae BY4741, a complete cytochrome P450 redox system was constructed. Through the combination of CYP68JX and heterologous CPRs, the yield of the target product 7α,15α-diOH-DHEA in CYP68JX recombinant system was increased to 37.8%. Furthermore, by adding NADPH coenzyme precursor tryptophan of 40 mmol/L and co-substrate fructose of 20 g/L during the conversion process, the catalytic efficiency of CYP68JX was further improved, the target product yield reached 57.9% which was 3.39-fold higher than initial yield. Overall, this study provides a reference for improving the catalytic activity of P450s. • CYP68JX achieved heterologous expression in Saccharomyces cerevisiae. • The combination of CYP68JX and AtCPR increased the catalytic efficiency. • Adding tryptophan and fructose improved NADPH supply. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 09600760
- Volume :
- 238
- Database :
- Academic Search Index
- Journal :
- Journal of Steroid Biochemistry & Molecular Biology
- Publication Type :
- Academic Journal
- Accession number :
- 175604216
- Full Text :
- https://doi.org/10.1016/j.jsbmb.2023.106452