Enzymes, Directed Evolution

Abstract

Directed evolution has emerged as a powerful method for engineering the catalytic profile of enzymes. It is based on repetitive cycles of random gene mutagenesis and expression coupled with efficient high‐throughput screening or selection for a given catalytic property such as thermostability, substrate acceptance, and/or enantioselectivity. In the 1990s, directed evolution was established on a broad front by applying standard mutagenesis methods such as error‐prone polymerase chain reaction (epPCR), saturation mutagenesis, and DNA shuffling. The current challenge is to devise optimal strategies for probing protein sequence space, thereby allowing for fast directed evolution. This article covers the progress of the last few years.