"Effective mutagenesis strategies in enzyme engineering are often dependent on the generation of small and targeted, high-quality libraries of mutants. Such 'smart' libraries are consistent with practical constraints imposed by the screening effort; while point-mutant libraries are readily screened, we need creative solutions to improve our capacity to explore the combinatorial complexity of sequence space."
In this webinar, Dr. Quaglia will talk about how she used a flexible approach based on directed evolution and the Golden Gate cloning strategy to target three distinct regions of Candida antarctica lipase A (Cal-A) generating a series of diverse libraries. Screening to improve discrimination of short-chain versus long-chain fatty acid substrates was aided by the development of a general, automated method that allowed for visual discrimination in the hydrolysis of varied substrates in whole cells.