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Use of green fluorescent protein and luciferase biomarkers to monitor survival and activity of Arthrobacter chlorophenolicus A6 cells during biodegradation of 4-chlorophenol in soil.
Stockholm University, Faculty of Science, Department of Biochemistry and Biophysics.
2001 In: Environmental Microbiology, Vol. 3, 32-42 p.Article in journal (Refereed) Published
Place, publisher, year, edition, pages
2001. Vol. 3, 32-42 p.
URN: urn:nbn:se:su:diva-23124OAI: diva2:190286
Part of urn:nbn:se:su:diva-159Available from: 2004-05-13 Created: 2004-05-13Bibliographically approved
In thesis
1. 4-chlorophenol biodegradation by Arthrobacter chlorophenolicus A6
Open this publication in new window or tab >>4-chlorophenol biodegradation by Arthrobacter chlorophenolicus A6
2004 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

A microorganism was isolated which could grow on unusually high concentrations of the toxic pollutant 4-chlorophenol. Taxonomic studies showed that the microorganism constituted a novel species within the genus Arthrobacter and it was named Arthrobacter chlorophenolicus A6.

A. chlorophenolicus A6 was chromosomally tagged with either the gfp gene, encoding the green fluorescent protein (GFP), or the luc gene, encoding firefly luciferase. When the tagged cells were inoculated into 4-chlorophenol contaminated soil they could completely remove 175 µg/g 4-chlorophenol within 10 days, whereas no loss of 4-chlorophenol was observed in the uninoculated control microcosms. During these experiments the gfp and luc marker genes allowed monitoring of cell number and metabolic status.

When A. chlorophenolicus A6 was grown on mixtures of phenolic compounds, the strain exhibited a preference for 4-nitrophenol over 4-chlorophenol, which in turn was preferred over phenol. Analysis of growth and degradation data indicated that the same enzyme system was used for removal of 4-chlorophenol and 4-nitrophenol. However, degradation of unbstituted phenol appeared to be mediated by another or an additional enzyme system. The luc-tagged A. chlorophenolicus A6 gave valuable information about growth, substrate depletion and toxicity of the phenolic compounds in substrate mixtures.

The 4-chlorophenol degradation pathway in A. chlorophenolicus A6 was elucidated. The metabolic intermediate subject to ring cleavage was found to be hydroxyquinol and two different pathway branches led from 4-chlorophenol to hydroxyquinol. A gene cluster involved in 4-chlorophenol degradation was cloned from A. chlorophenolicus A6. The cluster contained two functional hydroxyquinol 1,2-dioxygenase genes and a number of other open reading frames presumed to encode enzymes involved in 4-chlorophenol catabolism. Analysis of the DNA sequence suggested that the gene cluster had partly been assembled by horizontal gene transfer.

In summary, 4-chlorophenol degradation by A. chlorophenolicus A6 was studied from a number of angles. This organism has several interesting and useful traits such as the ability to degrade high concentrations of 4-chlorophenol and other phenols alone and in mixtures, an unusual and effective 4-chlorophenol degradation pathway and demonstrated ability to remove 4-chlorophenol from contaminated soil.

Place, publisher, year, edition, pages
Stockholm: Institutionen för biokemi och biofysik, 2004. 72 p.
National Category
Biochemistry and Molecular Biology
urn:nbn:se:su:diva-159 (URN)91-7265-875-4 (ISBN)
Public defence
2004-06-02, Magnélisalen, Kemiska övningslaboratoriet, Svante Arrhenius väg 12 A, Stockholm, 14:00
Available from: 2004-05-13 Created: 2004-05-13Bibliographically approved

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