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.