Directed Evolution of Protein Catalysts

Annu Rev Biochem. 2018 Jun 20:87:131-157. doi: 10.1146/annurev-biochem-062917-012034. Epub 2018 Mar 1.

Abstract

Directed evolution is a powerful technique for generating tailor-made enzymes for a wide range of biocatalytic applications. Following the principles of natural evolution, iterative cycles of mutagenesis and screening or selection are applied to modify protein properties, enhance catalytic activities, or develop completely new protein catalysts for non-natural chemical transformations. This review briefly surveys the experimental methods used to generate genetic diversity and screen or select for improved enzyme variants. Emphasis is placed on a key challenge, namely how to generate novel catalytic activities that expand the scope of natural reactions. Two particularly effective strategies, exploiting catalytic promiscuity and rational design, are illustrated by representative examples of successfully evolved enzymes. Opportunities for extending these approaches to more complex biocatalytic systems are also considered.

Keywords: computational design; enzyme catalysis; metalloenzymes; protein engineering; screening; selection.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • Biocatalysis
  • Directed Molecular Evolution / methods*
  • Drug Design
  • Enzymes / chemistry
  • Enzymes / genetics*
  • Enzymes / metabolism*
  • Genetic Variation
  • High-Throughput Screening Assays
  • Humans
  • Metabolic Networks and Pathways / genetics
  • Models, Molecular
  • Protein Engineering / methods
  • Proteins / chemistry
  • Proteins / genetics
  • Proteins / metabolism
  • Selection, Genetic
  • Stereoisomerism
  • Substrate Specificity

Substances

  • Enzymes
  • Proteins