A Bioengineered Stable Protein 1-Hemin Complex with Enhanced Peroxidase-Like Catalytic Properties

Enzymes have gained their unique efficiency and catalytic activity through billions of years of evolution, perfecting their active site to a desired reaction. Inspired by nature, a novel enzyme-mimicking platform is designed based on stable protein 1 (SP1) to create a nano-compartment that mimics peroxidase activity. The biohybrid reveals enhanced activity over the hemin cofactor alone and improved stability in organic solvents in comparison to native peroxidase. Furthermore, the utilization of the obtained biohybrid in an optical glucose biosensing platform is shown. The biohybrid crystallographic structure is solved, indicating that the SP1 structure is not affected by the hemin coordination. This work opens the path for developing new cofactor binding centers in engineered protein scaffolds for various artificial catalytic processes. Peroxidase is a heme-containing enzyme that has been broadly used in medical diagnosis, environmental treatments, and chemical syntheses. Its main limitation is low expression yields and poor stability in harsh environments. Herein, the feasibility and the use of Stable protein 1, a thermo-stable and stress-responsive protein, conjugated with hemin as a nano-compartment for peroxidase mimicry is shown.image (c) 2024 WILEY-VCH GmbH