(Reverse) Evolution of a Promiscuous Isochorismate Pyruvate Lyase into an Efficient Chorismate Mutase.

ABSTRACT: PchB is an isochorismate pyruvate lyase (IPL) involved in siderophore biosynthesis in Pseudomonas aeruginosa. Besides catalyzing the [1,5]-sigmatropic rearrangement of isochorismate, PchB also has weak chorismate mutase (CM) activity, promoting the [3,3]-sigmatropic transformation of chorismate. It has been suggested that the secondary metabolism enzyme PchB evolved from a primary metabolism CM precursor. Here, we employed directed evolution to convert PchB (back) into an efficient CM. A total of seven residues around the active site differing between PchB and a prototypical CM from Escherichia coli were randomized, and the resulting gene library was subjected to selection for CM activity. After growth selection in an auxotrophic strain, a catalyst with 10-fold

SUBMITTER: Kunzler DE 

PROVIDER: S-EPMC12329711 | biostudies-literature | 2025 Aug

REPOSITORIES: biostudies-literature