A Simple and Cost-Efficient Method for the Production of Recombinant Horseradish Peroxidase in E. coli

Horseradish peroxidase (HRP) is a chromogenic glycoenzyme widely used in research, diagnostics, and therapeutics. Due to its high demand, various eukaryotic and prokaryotic expression systems have been employed for the production of recombinant HRP. Eukaryotic systems yield properly folded, fully functional enzymes with the necessary post-translational modifications. However, these systems can be costly, time-consuming, and prone to hyperglycosylation. In contrast, prokaryotic systems are simple, inexpensive, and readily available, but achieving proper folding and subsequent modifications can be challenging. In this study, we employed a simple and cost-effective method to produce recombinant HRP in soluble form, using the E. coli expression system. The produced enzyme demonstrated substantial activity (89.75 ± 3.25 U/mg) and resistance to heat (T_1/2 = 5 min at 50 °C), pH variations (up to 8), and H₂O₂ concentrations (up to 10 mM). Additionally, we systematically compared our method with those of other researchers, highlighting methodological details and outcomes of HRP production in E. coli.