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Applied and Environmental Microbiology, October 1999, p. 4575-4581, Vol. 65, No. 10
Department of Biochemistry, Groningen
Biomolecular Sciences and Biotechnology Institute, University of
Groningen, 9747 AG Groningen, The Netherlands
Received 15 April 1999/Accepted 30 July 1999
Trihalogenated propanes are toxic and recalcitrant organic
compounds. Attempts to obtain pure bacterial cultures able to use these
compounds as sole carbon and energy sources were unsuccessful. Both the
haloalkane dehalogenase from Xanthobacter autotrophicus GJ10 (DhlA) and that from Rhodococcus sp. strain m15-3
(DhaA) were found to dehalogenate trihalopropanes to 2,3-dihalogenated propanols, but the kinetic properties of the latter enzyme are much
better. Broad-host-range dehalogenase expression plasmids, based on
RSF1010 derivatives, were constructed with the haloalkane dehalogenase
from Rhodococcus sp. strain m15-3 under the control of the
heterologous promoters Plac,
PdhlA, and Ptrc. The
resulting plasmids yielded functional expression in several
gram-negative bacteria. A catabolic pathway for trihalopropanes was
designed by introducing these broad-host-range dehalogenase expression plasmids into Agrobacterium radiobacter AD1, which has the
ability to utilize dihalogenated propanols for growth. The recombinant strain AD1(pTB3), expressing the haloalkane dehalogenase gene under the
control of the dhlA promoter, was able to utilize both 1,2,3-tribromopropane and 1,2-dibromo-3-chloropropane as sole carbon
sources. Moreover, increased expression of the haloalkane dehalogenase
resulted in elevated resistance to trihalopropanes.
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Utilization of Trihalogenated Propanes by Agrobacterium
radiobacter AD1 through Heterologous Expression of the Haloalkane
Dehalogenase from Rhodococcus sp. Strain m15-3
*
Corresponding author. Mailing address: Department of
Biochemistry, Groningen Biomolecular Sciences and Biotechnology
Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen,
The Netherlands. Phone: 31-50-363-4208. Fax: 31-50-363-4165. E-mail: d.b.janssen{at}chem.rug.nl.
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