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Applied and Environmental Microbiology, August 1999, p. 3641-3650, Vol. 65, No. 8
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Isolation and Characterization of Methanomethylovorans
hollandica gen. nov., sp. nov., Isolated from Freshwater
Sediment, a Methylotrophic Methanogen Able To Grow on Dimethyl
Sulfide and Methanethiol
Bart P.
Lomans,
Ronald
Maas,
Rianne
Luderer,
Huub
J. M.
Op den Camp,*
Arjan
Pol,
Chris
van der
Drift, and
Godfried D.
Vogels
Department of Microbiology and Evolutionary
Biology, Faculty of Science, University of Nijmegen, NL-6525 ED
Nijmegen, The Netherlands
Received 28 January 1999/Accepted 20 May 1999
A newly isolated methanogen, strain DMS1T, is the first
obligately anaerobic archaeon which was directly enriched and isolated from a freshwater sediment in defined minimal medium containing dimethyl sulfide (DMS) as the sole carbon and energy source. The use of
a chemostat with a continuous DMS-containing gas stream as a method of
enrichment, followed by cultivation in deep agar tubes, resulted in a
pure culture. Since the only substrates utilized by strain
DMS1T are methanol, methylamines, methanethiol (MT), and
DMS, this organism is considered an obligately methylotrophic
methanogen like most other DMS-degrading methanogens. Strain
DMS1T differs from all other DMS-degrading methanogens,
since it was isolated from a freshwater pond and requires NaCl
concentrations (0 to 0.04 M) typical of the NaCl concentrations
required by freshwater microorganisms for growth. DMS was degraded
effectively only in a chemostat culture in the presence of low hydrogen
sulfide and MT concentrations. Addition of MT or sulfide to the
chemostat significantly decreased degradation of DMS. Transient
accumulation of DMS in MT-amended cultures indicated that transfer of
the first methyl group during DMS degradation is a reversible process.
On the basis of its low level of homology with the most closely related methanogen, Methanococcoides burtonii (94.5%), its
position on the phylogenetic tree, its morphology (which is different
from that of members of the genera Methanolobus,
Methanococcoides, and Methanohalophilus), and
its salt tolerance and optimum (which are characteristic of freshwater
bacteria), we propose that strain DMS1T is a representative
of a novel genus. This isolate was named Methanomethylovorans
hollandica. Analysis of DMS-amended sediment slurries with a
fluorescence microscope revealed the presence of methanogens which were
morphologically identical to M. hollandica, as described in
this study. Considering its physiological properties, M. hollandica DMS1T is probably responsible for
degradation of MT and DMS in freshwater sediments in situ. Due to the
reversibility of the DMS conversion, methanogens like strain
DMS1T can also be involved in the formation of DMS through
methylation of MT. This phenomenon, which previously has been shown to
occur in sediment slurries of freshwater origin, might affect the
steady-state concentrations and, consequently, the total flux of DMS
and MT in these systems.
*
Corresponding author. Mailing address: Department of
Microbiology and Evolutionary Biology, Faculty of Science,
University of Nijmegen, Toernooiveld 1, NL-6525 ED Nijmegen, The
Netherlands. Phone: 31 (0) 24 3652657. Fax: 31 (0) 24 3652830. E-mail:
huubcamp{at}sci.kun.nl.
Applied and Environmental Microbiology, August 1999, p. 3641-3650, Vol. 65, No. 8
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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