High-Throughput Screening or Selection Methods for Evolutionary Enzyme Engineering

Abstract

Enzymes have become attractive candidates for the production ofindustrial chemicals and pharmaceutical intermediates comparedto conventional chemical catalysts. However, the properties ofnaturally existing enzymes are not always satisfactory for spe-cific functions. Tailoring and evolving enzyme characteristics byevolutionary enzyme engineering techniques is a powerful toolto create novel biocatalysts, but isolating variants with desiredproperties from a large pool of randomly generated mutants stillremains a bottleneck. Consequently, new selection and screeningmethodologies have been constantly developed to accomplish thegoals of evolutionary enzyme engineering. This review will focuson the recent development of novel high-throughput and ultrahigh-throughput selection and screening strategies to isolate enzymevariants from a large library of mutants.