Integrating continuous hypermutation with high-throughput screening for optimization of cis,cis-muconic acid production in yeast.

ABSTRACT: Directed evolution is a powerful method to optimize proteins and metabolic reactions towards user-defined goals. It usually involves subjecting genes or pathways to iterative rounds of mutagenesis, selection and amplification. While powerful, systematic searches through large sequence-spaces is a labour-intensive task, and can be further limited by a priori knowledge about the optimal initial search space, and/or limits in terms of screening throughput. Here, we demonstrate an integrated directed evolution workflow for metabolic pathway enzymes that continuously generate enzyme variants using the recently developed orthogonal replication system, OrthoRep and screens for optimal performance in high-throughput using a transcription factor-based biosensor. We demonstrate the strengths of this

SUBMITTER: Jensen ED 

PROVIDER: S-EPMC8601171 | biostudies-literature | 2021 Nov

REPOSITORIES: biostudies-literature