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Microsomal and cytosolic cytochrome P450 mediated benzo(a)pyrene hydroxylation in Pleurotus pulmonarius |
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| Authors: | S Masaphy D Levanon Y Henis K Venkateswarlu SL Kelly |
| Institution: | (1) MIGAL Technology Centre, 10200 Kiryat Shmona, Israel;(2) Hebrew University, 76100 Rehovot, Israel;(3) Department of Molecular Biology and Biotechnology, Krebs Institute for Biomolecular Research, Sheffield University, S102UH Sheffield, UK |
| Abstract: | Summary Microsomal and soluble fractions of Pleurotus pulmonarius exhibited a reduced carbon monoxide difference spectrum with P450 maxima at 448nm and 450–452nm respectively. Substrate induced Type I spectra were observed on addition of benzo(a)pyrene to both fractions. Benzo(a)pyrene hydroxylation was measured using the aryl hydrocarbon hydroxylase assay and was observed to be P450 dependent as indicated by carbon monoxide inhibition together with the substrate binding characteristics. The activity of the fractions were observed to give Km of 200mM and 660mM and Vmax of 1.25 nmol/min/nmol P450 and 0.57 nmol/min/nmol P450 for the microsomal and cytosolic fractions respectively. |
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