EAGER: Biocatalyst Orientation Control During Immobilization

Brigham Young University, Provo UT

Investigators

Abstract

Enzymatic biocatalysis is a particularly attractive technology in the effort to develop renewable, sustainable, and highly specific catalytic processes. The immobilization of enzymes on surfaces frequently leads to increased enzyme stability, decreased cost, and ease of enzyme separation and reuse. However, the available immobilization technologies result in random orientations of the immobilized enzyme which inhibits enzyme function. For optimal biocatalytic activity, the ability to control the enzyme attachment orientation is necessary to maximize the enzyme?s active site accessibility and minimize destabilizing enzyme-surface interactions. The PI, Bradley Bundy of Brigham Young University, Provo, UT has previously demonstrated the incorporation of certain unique unnatural amino acids into proteins at controlled locations anywhere in the protein with economically realistic high yields. This unique amino acid is used to covalently link proteins to each other using biocompatible click chemistry. The working hypothesis is that these same unique chemical moieties can be used with click chemistry to control the orientation of the immobilized enzyme on the surface to yield optimal orientation for activity and stability. Lipase B will be attached using this click chemistry to a magnetic substrate and tested for specific activity. This test will be performed for multiple attachment locations which by structure appear to make the active site accessible or inaccessible. The proposed work has the potential to scientifically impact a significant portion of the multibillion dollar catalytic enzyme industry. With its successful development this technology would lead to the transformation of the biocatalysis industry by providing a rapid, reliable, and covalent optimization method for immobilization. This work would also impact the protein-surface interaction field, providing researchers with immobilized enzymes that are all covalently attached in the same orientation. Properties tested from the bulk could then be reliably extrapolated to the atomic level. Sustainability would also be impacted as this work leads to cheaper, more efficient green biocatalysis to economically replace existing unsustainable and energy and waste intensive chemical synthesis methods.