oxidation of catechol in plants iv. purification and properties of the 3,4,3',4'-tetrahydroxydiphenyl-forming enzyme system from tecoma leaves

Kandaswami, C and Vaidyanathan, CS (1973) oxidation of catechol in plants iv. purification and properties of the 3,4,3',4'-tetrahydroxydiphenyl-forming enzyme system from tecoma leaves. In: Journal of Biological Chemistry, 248 (11). 4035 -4039.

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Abstract

Acetone powders prepared from leaf extracts of Tecoma stans L. were found to catalyze the oxidation of catechol to 3,4,3',4'-tetrahydroxydiphenyl. Fractionation of the acetone powders obtained from Tecoma leaves with acetone, negative adsorption of the acetone fraction with tricalcium phosphate gel, and chromatography of the gel supernatant on DEAE-Sephadex yielded a 68-fold purified enzyme with 66% recovery. The enzyme had an optimum pH around 7.2. It showed a temperature optimum of 30° and the Km for catechol was determined as 2 x 10-4 m. The purified enzyme moved as a single band on polyacrylamide gel electrophoresis. Its activity was found to be partially stimulated by Mg2+. The reaction was not inhibited by o-phenanthroline and agr,agr'-dipyridyl. The purified enzyme was highly insensitive to a range of copper-chelating agents. It was not affected appreciably by thiol inhibitors. The reaction was found to be suppressed to a considerable extent by reducing agents like GSH, cysteine, cysteamine, and ascorbic acid. The purified enzyme was remarkably specific for catechol. Catalase affected neither the enzyme activity nor the time course of the reaction. Hydrogen peroxide was not formed as a product of the reaction.

Item Type:	Journal Article
Publication:	Journal of Biological Chemistry
Publisher:	The American Society for Biochemistry and Molecular Biology
Additional Information:	Copy right of this article belongs to The American Society for Biochemistry and Molecular Biology.
Department/Centre:	Division of Biological Sciences > Biochemistry
Date Deposited:	07 Jan 2010 05:15
Last Modified:	19 Sep 2010 05:45
URI:	http://eprints.iisc.ac.in/id/eprint/23226

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