CAREER: Controlled Enzyme Biocatalyst Immobilization

Brigham Young University, Provo UT

Investigators

Abstract

INTELLECTUAL MERIT Enzymatic biocatalysis should be a major contributing technology as a foundation of renewable, sustainable and highly specific catalytic processes for the growing $600+ billion US chemical industry. The major challenges preventing widespread use of enzymatic biocatalysis are cost of production, limited stability and separation difficulties. While immobilization of enzymes to surfaces can address certain aspects of these challenges, traditional immobilization technologies do not allow the researcher to control and optimize the orientation of the attached enzyme on the surface. Enzyme orientation is less of concern for free enzymes in solution, but is of importance when they are fixed to a surface and restricted in motion. Dr. Bradley Bundy at Brigham Young University in Provo, Utah in this Faculty Early Career Development (CAREER) Program Award proposes to utilize site-specifically incorporated unnatural amino acids to directly control the orientation of the immobilized enzyme on a surface, thereby controlling the aspects exposed to reactants in solution. In order to accomplish the determination of the most suitable orientations, Dr. Bundy proposes to develop a high-throughput screen to rapidly test multiple orientations of the immobilized enzyme to optimize both enzyme activity and stability. With data on orientation being generated, a further objective of this work is to gather orientation-specific immobilized enzyme stability and activity data in bulk for processing and analysis. This data will enable the validation of molecular interaction simulations and also assist in the development of design guidelines to predict in advance orientations of immobilized enzymes which both improve enzyme stability and function. BROAD IMPACT The proposal includes an integrated research and education plan with multiple levels of impact. The successful development of the proposed technology has the potential to significantly impact biocatalysis, medical diagnosis, biohazard sensing, proteomics and any application that uses proteins immobilized on surfaces. The award work promotes sustainability as it develops less expensive, more efficient green biocatalysis to economically replace existing unsustainable and energy and waste intensive chemocatalytic processes. The education component of Dr. Bundy?s proposal addresses the U.S. President?s Council on Jobs and Competitiveness goal of producing 1 million additional college graduates from Science, Technology, Engineering and Mathematics (STEM). Towards this aim, the proposal seeks to light the fire of discovery in students early and maintain it through graduate school. Elementary students will be introduced to enzymes and biocatalysis through fun, grade-individualized, hands-on teaching modules that are feedback-optimized by the PI and leveraged for impact through university engineering club participation and community science events. A STEM-promoting learning environment for grades 7-12 students will be created to meet the strong need for biotechnology education materials and teacher-training. To maximize impact and distribution, the PI will collaborate with the NSF-sponsored National Center for Engineering and Technology Education at Brigham Young University which directly works with teachers state-wide. The award is being jointly made between the CBET Division's Catalysis & Biocatalysis Program and the NSF EPSCoR Program.