Structure-based Engineering of a Plant-Fungal Hybrid Peroxidase for Enhanced Temperature and pH Tolerance

In an age of ever-increasing biotechnological and industrial demand for new and specialized biocatalysts, rational protein engineering offers a direct approach to enzyme design and innovation. Heme peroxidases, as indispensable oxidative biocatalysts, provide a relatively mild alternative to the traditional harsh, and often toxic, chemical catalysts, and subsequently, have found widespread application throughout industry. However, the potential for these enzymes is far greater than their present use, as processes are currently restricted to the more stable, but less catalytically powerful, subset of peroxidases. Here we describe the structure-guided, rational engineering of a plant-fungal hybrid peroxidase built to overcome the application barrier of these high-reduction potential peroxidases. This engineered enzyme has the catalytic versatility and oxidative ability of a high-reduction potential versatile peroxidase, with enhanced temperature and pH tolerance similar to that of a highly stable plant peroxidase. [Display omitted] •Structure-based engineering enables design of peroxidase with wider utility•Catalytic versatility of engineered peroxidase (VP2.0) comparable with catalytic parent•VP2.0 retains activity over broader temperature and pH ranges•VP2.0 exhibits greater structural stability under optimal reaction conditions Kohler et al. describe a rational engineering approach for the direct design of innovative, industrially tailored biocatalysts. They utilize this approach to address the current application barrier of high-reduction potential peroxidases as oxidative tools, building a catalytically versatile plant-fungal hybrid peroxidase capable of functioning under wider temperature and pH ranges.