Growth-coupled continuous directed evolution by MutaT7 enables efficient and automated enzyme engineering.

ABSTRACT: Traditional directed evolution is limited by labor-intensive iterative steps and low-throughput selection and screening. To address these challenges, we developed a growth-coupled continuous directed evolution (GCCDE) approach, enabling automated and efficient enzyme engineering. By linking enzyme activity to bacterial growth and utilizing the MutaT7 system, GCCDE combines in vivo mutagenesis and high-throughput selection of superior enzyme variants in a single process. To validate this approach, we evolved the thermostable enzyme CelB from Pyrococcus furiosus to enhance its β-galactosidase activity at lower temperatures while maintaining thermal stability. CelB activity was coupled to the growth of E. coli, allowing variants with improved activity to utilize lactose m

SUBMITTER: Deng Y 

PROVIDER: S-EPMC12016552 | biostudies-literature | 2025 Apr

REPOSITORIES: biostudies-literature