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A novel 55-kDa hydroxylase was isolated from cultured cells of Catharanthus roseus by a three-step procedure: anion exchange chromatography, affinity chromatography and hydroxylapatite adsorption chromatography. The enzyme specifically catalyzed the hydroxylation of 2-hydroxybenzoic acid to give 2,5-dihydroxybenzoic acid. The enzyme activity was optimal at pH 7.8 and was completely inhibited by divalent...
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