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Degradation of 2-chloroethanol by wild type and mutants of Pseudomonas putida US2 |
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| Authors: | Uwe J Strotmann Marjan Pentenga Dick B Janssen |
| Institution: | (1) Institute of Microbiology, University of Münster, Corrensstrasse 3, D-4400 Münster, Federal Republic of Germany;(2) Department of Biochemistry, Groningen Biotechnology Centre, University of Groningen, Nijenbrogh 16, 9747 AG Groningen, The Netherlands;(3) Emissionsüberwachung und Ökologie, BASF AG, D-6700 Ludwigshafen, Federal Republic of Germany |
| Abstract: | A strain of Pseudomonas putida was isolated that was able to degrade 2-chloroethanol. The degradation proceeded via 2-chloroacetaldehyde and chloroacetate to glycolate. In crude extracts the enzymes for this degradation pathway could be detected. All enzymes proved to be inducible. The dehalogenase that catalyzed the dehalogenation of chloroacetate to glycolate was further characterized. It consisted of a single polypeptide chain with a molecular mass of 28 kDa. After induction the dehalogenase was expressed at a high level. In a mutant resistant to high concentrations of 2-chloroethanol the dehalogenase was no longer expressed. The mechanism of resistance seemed to be due to the inability to convert chloroacetate and export of this compound out of the cell.Non-standard abbreviations CEO 2-chloroethanol - DCPIP 2,6-dichlorophenolindophenol - FPLC fast protein liquid chromatography - PAGE polyacrylamide gelelectrophoresis - PES phenazine ethosulfate - PMS phenazine methosulfate - PQQ pyrroloquinoline quinone |
| Keywords: | Chloroethanol degradation Pseudomonas putida Degradative enzymes Dehalogenase Chloroethanol resistance |
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