Orthogonal replication with optogenetic selection evolves yeast JEN1 into a mevalonate transporter.

ABSTRACT: The in vivo continuous evolution system OrthoRep (orthogonal replication) is a powerful strategy for rapid enzyme evolution in Saccharomyces cerevisiae that diversifies genes at a rate exceeding the endogenous genome mutagenesis rate by several orders of magnitude. However, it is difficult to neofunctionalize genes using OrthoRep partly because of the way selection pressures are applied. Here we combine OrthoRep with optogenetics in a selection strategy we call OptoRep, which allows fine-tuning of selection pressure with light. With this capability, we evolved a truncated form of the endogenous monocarboxylate transporter JEN1 (JEN1t) into a de novo mevalonate importer. We demonstrate the functionality of the evolved JEN1t (JEN1t^Y180C/G) in the production of farnesene, a renewab

SUBMITTER: Wegner SA 

PROVIDER: S-EPMC12405511 | biostudies-literature | 2025 Sep

REPOSITORIES: biostudies-literature