: Biocatalysts play crucial roles in the construction of therapeutics, and have the additional potential to serve as novel classes of therapeutics by modifying disease-associated proteins. Despite their significant advantages, the use of enzyme catalysts in industry and academic research has been limited by their narrow substrate scope. Directed evolution methods have demonstrated the ability to engineer specific enzymes to adopt new functions not present in the wild-type enzymes, however, conventional laboratory evolution strategies are time- consuming and their application to a variety of new enzyme functions has proven challenging. Here I propose two approaches to evolve bond-forming catalysts, using phage-assisted continuous evolution (PACE) or yeast display-based bond-formation selection, two powerful directed evolution platforms recently developed by the Liu group. The evolution of bond-forming catalysts will permit the discovery of new enzymes with tailor-made substrate scopes. As an initial application of these methods and concepts, we are evolving variants of sortase, a transamidase enzyme, to catalyze bond-formation events with tailor-made specificities, including specificities for disease-associated proteins. Our preliminary results suggest the successful evolution of bond-forming enzymes with novel substrate specificities.

Public Health Relevance

Biocatalysts are vitally important in the construction of therapeutics in both academic and industrial laboratories, but are frequently limited to catalyzing reactions that take place in nature. I propose to use continuous directed evolution to rapidly generate enzymes with new, tailor-made function that can facilitate the synthesis of therapeutically relevant small molecules.

Agency
National Institute of Health (NIH)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
1F32GM109686-01A1
Application #
8783648
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Barski, Oleg
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Harvard University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
City
Cambridge
State
MA
Country
United States
Zip Code
02138