In vitro reconstitution of the shikimate pathway: a new tool for probing MST enzyme mechanisms

Abstract

"The MST enzymes (menaquinone, siderophore, tryptophan) are structurally homologous proteins that utilize chorismate in the biosynthetic pathways that generate aromatic amino acids, folate, quinones, and siderophores. Inhibitors of these enzymes could serve as new antibiotics. To help inform inhibitor design our lab will measure kinetic isotope effects in MST enzymes using isotopically labeled chorismate. Chorismate is naturally synthesized in bacteria, fungi, and plants with the seven-enzyme shikimate pathway. Here we report the in vitro reconstitution of the shikimate pathway utilizing heterologously expressed shikimate enzymes combined with substrates and cofactors in a one-pot reaction to generate chorismate."