Bioactive expression of eukaryotic cytochrome P450 ferulate-5-hydroxylase in Escherichia coli for sustainable synthesis of antioxidant 5-hydroxyferulic acid
The eukaryotic cytochrome P450 ferulate-5-hydroxylase (F5H), a membrane-bound protein, plays a critical role in lignin synthesis involved in the biosynthesis of 5-Hydroxyferulic acid (5-HFA) from ferulic acid, with 5-HFA offering enhanced antioxidant properties. In this work, F5H derived from Arabidopsis thaliana (AtF5H) and NADPH-dependent cytochrome P450 reductase (CPR) were co-expressed in E. coli for the synthesis of ortho-hydroxylated ferulic acid. Simultaneously, by modifying the N-terminal regions of membrane proteins, the concentration of product 5-HFA could be increased up to 63.6 mg/L. Ferulic acid was extracted from discarded agricultural by-products, demonstrating a sustainable approach to valorizing agricultural waste. This study not only provides a valuable reference for the expression of plant-derived P450s in E. coli but also highlights the potential of these engineered strains to effectively utilize biological resources, offering economic benefits.