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Directed Evolution Provides Insight Into Conformational Substrate Sampling By Srta, Muna Suliman, Vishaka Santosh, Tom C. M. Seegar, Annamarie C. Dalton, Kathryn M. Schultz, Candice S. Klug, William A. Barton

Biochemistry and Molecular Biology Publications

The Sortase family of transpeptidases are found in numerous gram-positive bacteria and involved in divergent physiological processes including anchoring of surface proteins to the cell wall as well as pili assembly. As essential proteins, sortase enzymes have been the focus of considerable interest for the development of novel anti-microbials, however, more recently their function as unique transpeptidases has been exploited for the synthesis of novel bio-conjugates. Yet, for synthetic purposes, SrtA-mediated conjugation suffers from the enzyme's inherently poor catalytic efficiency. Therefore, to identify SrtA variants with improved catalytic efficiency, we used directed evolution to select a catalytically enhanced SrtA enzyme. …