Chorismic Acid → 1-Carboxy-4-hydroxy-alpha-oxo-2,5-cyclohexadiene-1-propanoic acid
Reaction equation
C=C(O[C@H:7]1[C@H:12]([OH:13])[CH:11]=[CH:10][C:9]([C:14]([OH:16])=[O:15])=[CH:8]1)C(O)=O.C1([CH2:23][C:24](=[O:28])[C:25]([O-:27])=[O:26])C=CC=CC=1>>[CH:11]1[CH:12]([OH:13])[CH:7]=[CH:8][C:9]([C:14]([OH:16])=[O:15])([CH2:23][C:24]([C:25]([OH:27])=[O:26])=[O:28])[CH:10]=1Participants
3
Procedure
One assay commonly used to measure chorismate mutase activity is based on the spectrophotometric estimation of phenylpyruvate obtained by acid conversion of the reaction product of prephenate. Metzenberg and Mitchell (1954) Arch Biochem and Biophys 243:374. In this assay, phenylpyruvate is estimated by the absorbance of either the enol-tautomer in alkaline solution (E=17,500 M−1 cm−1 at 320 nm in 1 M NaOH) or the enol-borate complex in either a concentrated solution of sodium borate and arsenate or phosphate (E=9292 M−1 cm−1 at 300 nm). Alternatively, the disappearance of chorismate may be assayed spectrophotometrically. Nishioka and Woodin (1972) Anal Biochem 45:617. The details of these assays and their advantages and disadvantages are described in Gilchrist and Connelly (1987) Methods in Enzymology 142:450-463.