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Journal of Bacteriology, April 2000, p. 1956-1963, Vol. 182, No. 7
Department of Biochemistry, Groningen
Biomolecular Sciences and Biotechnology Institute, University of
Groningen, NL-9747 AG Groningen, The Netherlands
Received 29 September 1999/Accepted 30 December 1999
The genes involved in isoprene (2-methyl-1,3-butadiene) utilization
in Rhodococcus sp. strain AD45 were cloned and
characterized. Sequence analysis of an 8.5-kb DNA fragment showed the
presence of 10 genes of which 2 encoded enzymes which were previously
found to be involved in isoprene degradation: a glutathione
S-transferase with activity towards
1,2-epoxy-2-methyl-3-butene (isoI) and a 1-hydroxy-2-glutathionyl-2-methyl-3-butene dehydrogenase
(isoH). Furthermore, a gene encoding a second glutathione
S-transferase was identified (isoJ). The
isoJ gene was overexpressed in Escherichia coli
and was found to have activity with 1-chloro-2,4-dinitrobenzene and
3,4-dichloro-1-nitrobenzene but not with 1,2-epoxy-2-methyl-3-butene. Downstream of isoJ, six genes (isoABCDEF) were
found; these genes encoded a putative alkene monooxygenase that showed
high similarity to components of the alkene monooxygenase from
Xanthobacter sp. strain Py2 and other multicomponent
monooxygenases. The deduced amino acid sequence encoded by an
additional gene (isoG) showed significant similarity with
that of
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Copyright © 2000, American Society for Microbiology. All rights reserved.
Characterization of the Gene Cluster Involved in
Isoprene Metabolism in Rhodococcus sp. Strain AD45
-methylacyl-coenzyme A racemase. The results are in
agreement with a catabolic route for isoprene involving epoxidation by
a monooxygenase, conjugation to glutathione, and oxidation of the
hydroxyl group to a carboxylate. Metabolism may proceed by fatty acid
oxidation after removal of glutathione by a still-unknown mechanism.
*
Corresponding author. Mailing address: Department of
Biochemistry, Groningen Biomolecular Sciences and Technology Institute, University of Groningen, Nijenborgh 4, NL-9747 AG Groningen, The Netherlands. Phone: 31-50-3634209. Fax: 31-50-3634165. E-mail: d.b.janssen{at}chem.rug.nl.
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