Enzymatic Stetter Reaction: Computational Study of the Reaction Mechanism of MenD

Quantum chemical calculations are used to investigate the detailed reaction mechanism of 2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylic-acid (SEPHCHC) synthase (also known as MenD), a thiamin diphosphate-dependent decarboxylase that catalyzes the formation of SEPHCHC from 2-ketoglutarate and isochorismate. This enzyme is involved in the menaquinone biosynthesis pathway in M. tuberculosis and is thought of as a potential drug target for anti-tuberculosis therapeutics. In addition, MenD shows promise as a biocatalyst for the synthesis of 1,4-functionalized compounds. Models of the active site are constructed on the basis of available X-ray structures, and the intermediates and transition states involved in the reaction mechanism are optimized and characterized. The calculated mechanism is in good agreement with prior kinetic studies and gives new insights into the mode of action of the enzyme. In particular, the structure and role of the tetrahedral post-decarboxylation intermediate observed in X-ray structures are discussed.