Design and Characterization of Combinatorial Libraries of Proteins

Abstract

With the recent development of de novo enzymes which retain activity both in vitro and in vivo there is a potential to use their specific sequences for the development of other de novo enzymes with similar properties, additionally, further investigation into de novo enzymes’ properties may lead to better rational design in the future. The fertility of a rationally-designed combinatorial Syn-F4-based library has been assayed on several auxotroph strains, leading to the rescue of several, including ones whose native enzymes use PLP, and ones whose native mechanisms vary greatly from the esterase function of Syn-F4. Investigation into the active geometry of Syn-F4 reveals the difficulty in probing protein geometries.