Evolving the naturally compromised chorismate mutase from Mycobacterium tuberculosis to top performance.

ABSTRACT: Chorismate mutase (CM), an essential enzyme at the branch-point of the shikimate pathway, is required for the biosynthesis of phenylalanine and tyrosine in bacteria, archaea, plants, and fungi. MtCM, the CM from Mycobacterium tuberculosis, has less than 1% of the catalytic efficiency of a typical natural CM and requires complex formation with 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase for high activity. To explore the full potential of MtCM for catalyzing its native reaction, we applied diverse iterative cycles of mutagenesis and selection, thereby raising k_cat/K_m 270-fold to 5 × 10⁵m^-1s^-1, which is even higher than for the complex. Moreover, the evolutionarily optimized autonomous MtCM, which had 11 of its 90 amino acids exc

SUBMITTER: Fahrig-Kamarauskait J 

PROVIDER: S-EPMC7762937 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature