Recombinant Human Myeloperoxidase from Pichia pastoris: Functional Analyses and Potential for Surface Applications

Human myeloperoxidase (hMPO) catalyzes the conversion of hydrogen peroxide and chloride into hypochlorous acid and plays a key role in human immunity. While commercial MPO is typically purified from human blood or produced recombinantly in mammalian cells or E. coli , we present novel approach for producing active recombinant hMPO (rhMPO) in Pichia pastoris . To eliminate the need for enzyme coupling, we engineered a chimeric protein by combining glucose oxidase from Penicillium amagasakiense and rhMPO. The two open reading frames were fused by PCR, yielding a bifunctional enzyme capable of generating hypochlorous acid directly from glucose. Additionally, the enzymes were immobilized on silicone model substrates via adsorption from solution to generate surface activity. This yeast-based production system provides a cost-effective and scalable platform for large-scale MPO synthesis, opening the development of novel biocatalytic applications.