Immobilization of enzymes is required for most biocatalytic processes, but chemistries used in enzyme immobilization are limited and can be challenging. Genetically encoded protein-based biomaterials could provide easy-to-use immobilization platforms for biocatalysts. We recently developed a self-assembling protein scaffold that covalently immobilized SpyTagged enzymes by engineering the bacterial microcompartment protein EutM from Salmonella enterica with a SpyCatcher domain. We also identified a range of EutM homologues as robust protein nanostructures with diverse architectures and electrostatic surface properties. In this work, we created a modular immobilization platform with tunable surface properties by developing a toolbox of self-assembling, robust EutM-SpyCatcher scaffolds. Using an alcohol dehydrogenase as model biocatalyst, we show that the scaffolds improve enzyme activity and stability. This work provides a modular, easy-to-use immobilization system that can be tailored for the optimal function of biocatalysts of interest.
Bibliographical noteFunding Information:
TEM imaging of scaffolds was conducted using equipment provided by University Imaging Center, University of Minnesota. This work was supported by funds provided by Defense Threat Reduction Agency Grant HDTRA1-15-0004 and Defense Advanced Research Projects Agency Contract HR0011-17-2-0038. T.J. was supported by funding from a Grand Challenge research award from the University of Minnesota.
- alcohol dehydrogenase
- protein scaffolds
- synthetic biology