Bulk purification of isochorismic acid by low-pressure octadecyl (C18) reverse-phase liquid chromatography.
Partially purified isochorismate synthase (EC 5.4.99.6) from Klebsiella pneumoniae 62-1 was used to produce bulk quantities (3.4-6.8 mg) of isochorismate from chorismate. A new, preparative, low-pressure liquid chromatographic method for the purification of isochorismate was used; a (1.0 X 13.0 cm) octadecyl (C18) reverse-phase column with a discontinuous, stepped methanol gradient as eluent. The recovery of isochorismate was quantitative and its purity was verified by HPLC using a butyl (C4) reverse-phase column. This chromatographic method is superior to those previously described.
Seeger
J W
JW
Department of Biological Sciences, University of Pittsburgh, Pennsylvania 15260.
Bentley
R
R
D002827:Chorismic Acid / Q000302:isolation & purification*; D002853:Chromatography, Liquid / Q000379:methods*; D053138:Cyclohexenes; D019751:Intramolecular Transferases; D007535:Isomerases / Q000494:pharmacology; D007536:Isomerism; D007711:Klebsiella pneumoniae / Q000201:enzymology
D053138:Cyclohexenes; C052985:isochorismic acid; D007535:Isomerases; D019751:Intramolecular Transferases; C058486:isochorismate synthase; D002827:Chorismic Acid