Experimental and computational investigation of the uncatalyzed rearrangement and elimination reactions of isochorismate.
The versatile biosynthetic intermediate isochorismate decomposes in aqueous buffer by two competitive pathways, one leading to isoprephenate by a facile Claisen rearrangement and the other to salicylate via elimination of the enolpyruvyl side chain. Computation suggests that both processes are concerted but asynchronous pericyclic reactions, with considerable C-O cleavage in the transition state but relatively little C-C bond formation (rearrangement) or hydrogen atom transfer to the enolpyruvyl side chain (elimination). Kinetic experiments show that rearrangement is roughly 8-times more favorable than elimination. Moreover, transfer of the C2 hydrogen atom to C9 was verified by monitoring the decomposition of [2-(2)H]isochorismate, which was prepared chemoenzymatically from labeled shikimate, by (2)H NMR spectroscopy and observing the appearance of [3-(2)H]pyruvate. Finally, the isotope effects obtained with the C2 deuterated substrate are in good agreement with calculations assuming pericyclic reaction mechanisms. These results provide a benchmark for mechanistic investigations of isochorismate mutase and isochorismate pyruvate lyase, the enzymes that respectively catalyze the rearrangement and elimination reactions in plants and bacteria.
DeClue
Michael S
MS
Laboratory of Organic Chemistry, ETH Zürich, CH-8093 Zürich, Switzerland.
Baldridge
Kim K
KK
Kast
Peter
P
Hilvert
Donald
D
D002826:Chorismate Mutase / Q000737:chemistry / Q000378:metabolism; D002827:Chorismic Acid / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D053138:Cyclohexenes; D003903:Deuterium / Q000737:chemistry; D006358:Hot Temperature; D007553:Isotope Labeling; D007700:Kinetics; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D007652:Oxo-Acid-Lyases / Q000737:chemistry / Q000378:metabolism
D053138:Cyclohexenes; C052985:isochorismic acid; D003903:Deuterium; D007652:Oxo-Acid-Lyases; C075960:chorismate pyruvate lyase; D002826:Chorismate Mutase; D002827:Chorismic Acid