"Multiagent" Screening Improves Directed Enzyme Evolution by Identifying Epistatic Mutations.

Journal Article (Journal Article)

Enzyme evolution has enabled numerous advances in biotechnology and synthetic biology, yet still requires many iterative rounds of screening to identify optimal mutant sequences. This is due to the sparsity of the fitness landscape, which is caused by epistatic mutations that only offer improvements when combined with other mutations. We report an approach that incorporates diverse substrate analogues in the screening process, where multiple substrates act like multiple agents navigating the fitness landscape, identifying epistatic mutant residues without a need for testing the entire combinatorial search space. We initially validate this approach by engineering a malonyl-CoA synthetase and identify numerous epistatic mutations improving activity for several diverse substrates. The majority of these mutations would have been missed upon screening for a single substrate alone. We expect that this approach can accelerate a wide array of enzyme engineering programs.

Full Text

Duke Authors

Cited Authors

  • Yang, T; Ye, Z; Lynch, MD

Published Date

  • May 4, 2022

Published In

Volume / Issue

  • 11 / 5

Start / End Page

  • 1971 - 1983

PubMed ID

  • 35507897

Electronic International Standard Serial Number (EISSN)

  • 2161-5063

International Standard Serial Number (ISSN)

  • 2161-5063

Digital Object Identifier (DOI)

  • 10.1021/acssynbio.2c00136


  • eng