Mechanoism o the salicylate hydroxylase reaction III. Characterization and reactivity of chemically or photochemically reduced enzyme-flavin

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  • Mechanism of the salicylate hydroxylase reaction

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Abstract 1. 1.|The spectral natures of chemically- and photochemically-reduced salicylate hydroxylase (salicylate, NADH: oxygen oxidoreductase (1-hydroxylating, 1-decarboxylating)) from Pseudomonas putida have been investigated. 2. 2.|When salicylate hydroxylase in the presence of substrate was titrated with a limited amount of dithionite or was illuminated in the presence of EDTA, a new species with a weak absorption band at a long wavelength appeared, whereas an entirely different species which was typical for the “red flavoprotein radical” was detected in the absence of substrate. The latter was converted to the former upon anaerobic addition of substrate. 3. 3.|Reoxidation of the reduced enzyme-substrate complex with air formed the product in an amount stoichiometric with the reduced flavin.

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