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Appl Environ Microbiol, April 1998, p. 1194-1202, Vol. 64, No. 4
Mikrobiologisches Institut, ETH Zürich,
ETH-Zentrum, CH-8092 Zürich, Switzerland
Received 13 October 1997/Accepted 14 January 1998
Methylobacterium sp. strain DM4 and
Methylophilus sp. strain DM11 can grow with dichloromethane
(DCM) as the sole source of carbon and energy by virtue of homologous
glutathione-dependent DCM dehalogenases with markedly different kinetic
properties (the kcat values of the enzymes of
these strains are 0.6 and 3.3 s
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Effects of Bacterial Host and Dichloromethane
Dehalogenase on the Competitiveness of Methylotrophic
Bacteria Growing with Dichloromethane
1, respectively, and the
Km values are 9 and 59 µM, respectively). These strains, as well as transconjugant bacteria expressing the DCM
dehalogenase gene (dcmA) from DM11 or DM4 on a
broad-host-range plasmid in the background of dcmA mutant
DM4-2cr, were investigated by growing them under growth-limiting
conditions and in the presence of an excess of DCM. The maximal growth
rates and maximal levels of dehalogenase for chemostat-adapted bacteria
were higher than the maximal growth rates and maximal levels of
dehalogenase for batch-grown bacteria. The substrate saturation
constant of strain DM4 was much lower than the
Km of its associated dehalogenase, suggesting
that this strain is adapted to scavenge low concentrations of DCM.
Strains and transconjugants expressing the DCM dehalogenase from strain
DM11, on the other hand, had higher growth rates than bacteria
expressing the homologous dehalogenase from strain DM4. Competition
experiments performed with pairs of DCM-degrading strains revealed that
a strain expressing the dehalogenase from DM4 had a selective advantage
in continuous culture under substrate-limiting conditions, while
strains expressing the DM11 dehalogenase were superior in batch culture
when there was an excess of substrate. Only DCM-degrading bacteria with
a dcmA gene similar to that from strain DM4, however, were
obtained in batch enrichment cultures prepared with activated sludge
from sewage treatment plants.
*
Corresponding author. Mailing address:
Mikrobiologisches Institut, ETH Zürich, ETH-Zentrum/LFV, CH-8092
Zürich, Switzerland. Phone: 41 1 632 33 57. Fax: 41 1 632 11 48. E-mail: svuilleu{at}micro.biol.ethz.ch.
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