This project focuses on renewable biocatalysts for the efficient degradation of persistent organic contaminants in water. Surface-display enzyme biocatalysts are a new class of biocatalysts that have recently been shown to have high catalytic efficiency and high stability. With surface display technology, functional enzymes are expressed and displayed on the outer surface of microbial host cells by fusion with a cell wall using molecular biology techniques. Research efforts will be integrated with an education plan that impacts education at multiple levels, ranging from middle school to graduate school. The success of the proposed research will enable development of more effective tools to implement biocatalysis using synthetic biology for addressing pressing challenges of water sustainability. In future work use of other target enzymes in the cell surface-display system will be used to create new biocatalysts to address challenges relevant to other components at the food-water-energy nexus.
The overall research goal of this project is to develop a new class of engineered biocatalysts with surface display technology for the effective degradation of persistent organic contaminants. Initially, fungal enzymes will be used in the investigation of cell surface-display systems to harness enzyme activities with a focus on degradation of endocrine disrupting compounds. Renewability is assured by the ability to generate and automatically localize the target enzyme on the surface of cells through engineered biological machinery, making production and regeneration of the biocatalyst efficient and scalable. The research plan will address fundamental aspects: i) molecular design of surface display machinery for high catalytic efficiency, ii) a strategy for improving robustness under conditions relevant to advanced wastewater treatment, iii) creation of multi-enzyme surface displays for efficient cascade reactions. Renewable biocatalysts with different functions will be created and applied to biotransformations beyond contaminant degradation. Research efforts will be integrated with an education plan that aims to i) increase the motivation and participation of female and other underrepresented students in pursuing careers in environmental engineering and science, ii) educate the next generation of engineers regarding synthetic biology for environmental sustainability, and iii) enhance interdisciplinary thinking and communication skills in graduate training. These goals will be addressed in synergy with the research by leading undergraduates in both research and outreach, 6-12 grade outreach, new course development, and organization of an interdisciplinary graduate symposium.
