Abstract
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 × 13.0 cm) octadecyl (C 18) 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 (C 4) reverse-phase column. This chromatographic method is superior to those previously described.
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