Directed evolution of crotonyl-CoA reductase for increased solubility
Biofuels have received much attention lately as the need for a renewable and carbon-neutral source of energy becomes increasingly important. We have assembled a biosynthetic pathway in Escherichia coli that produces butanol, a second generation biofuel, in six overall steps. However, there is a significant bottleneck at crotonyl-CoA reductase (Ccr) in the pathway that limits butanol production, which may be attributed to the observation that Ccr suffers from low solubility. The overall goal of my project is to carry out directed evolution of Ccr and generate a library of mutants that can be screened for increased solubility and assayed for improved activity. In the future, the mutants generated in this work will help us to understand fundamental issues related to pathway flux as well as increase production of butanol in engineered E. coli.
Message to Sponsor
- Major: Molecular and Cell Biology
- Mentor: Michelle Chang, Chemistry