Purification and some properties of (1R,2S)-1,2-dihydroxy-3,5-cyclohexadiene-1,4-dicarboxylate dehydrogenase from Comamonas testosteroni T-2 |
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Authors: | ElisabethSalier HeikeLaue Hans R SchläfliOppenberg Alasdair MCook |
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Institution: | Institute of Microbiology, Swiss Federal Institute of Technology, ETH-Zentrum, CH-8092 Zürich, Switzerland; Faculty of Biology, University of Konstanz, PO Box 55 60, D-78434 Konstanz, Germany |
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Abstract: | Abstract Inducible (1 R ,2 S )-1,2-dihydroxy-3,5-cyclohexadiene-l,4-dicarboxylate (diene-diol) dehydrogenase was found in extracts of Comamonas testosteroni T-2 grown in p -toluate-or terephthalate-salts medium and it was purified using anion exchange, hydrophobic interaction and gel filtration chromatography. The enzyme is a homodimer with subunit M r 39000. It had a specific activity of 500 mkat/kg of protein and was activated by the addition of Fe2+. The dehydrogenase converted 1 mol diene-diol and 1 mol NAD+ to 1 mol protocatechuic acid, 1 mol NADH and 1 mol CO2. Apparent K m-values of 43 μM (NAD+) and about 90 μM (diene-diol) were determined. The hydride ion was transferred to the si face of NAD+. |
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Keywords: | Terephthalate Diene-diol dehydrogenase Enzyme purification Comamonas testosteroni T-2 Biodegradation |
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