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Cell extracts of toluene-grown Pseudomonas putida produced a soluble yellow dye during aerobic incubations with indole and NADH. Accumulation of indoxyl in reaction mixtures corresponded with a linear increase in absorbance at 400 nm. The rate of increase in absorbance was shown to be a specific measure of toluene dioxygenase activity. The primary product of toluene oxidation, cis-toluene dihydrodiol, inhibited dioxygenase activity in cell extracts containing no detectable activity of cis-toluene dihydrodiol dehydrogenase.
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Jenkins et al. (1985) studied this question.
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