Affiliation:
1. Department of Biological Sciences, University of Warwick, Coventry CV4 7AL
2. IBMC-Universidade do Porto R. Campo Alegre, 823 4150-180 Porto, Portugal
3. Division of Life Sciences, King’s College London, London SE1 9NN, United Kingdom
Abstract
ABSTRACT
Marinosulfonomonas methylotropha
strain TR3 is a marine methylotroph that uses methanesulfonic acid (MSA) as a sole carbon and energy source. The genes from
M. methylotropha
strain TR3 encoding methanesulfonate monooxygenase, the enzyme responsible for the initial oxidation of MSA to formaldehyde and sulfite, were cloned and sequenced. They were located on two gene clusters on the chromosome of this bacterium. A 5.0-kbp
Hin
dIII fragment contained
msmA, msmB
, and
msmC
, encoding the large and small subunits of the hydroxylase component and the ferredoxin component, respectively, of the methanesulfonate monooxygenase, while a 6.5-kbp
Hin
dIII fragment contained duplicate copies of
msmA
and
msmB
, as well as
msmD
, encoding the reductase component of methanesulfonate. Both sets of
msmA
and
msmB
genes were virtually identical, and the derived
msmA
and
msmB
sequences of
M. methylotropha
strain TR3, compared with the corresponding hydroxylase from the terrestrial MSA utilizer
Methylosulfonomonas methylovora
strain M2 were found to be 82 and 69% identical. The
msmA
gene was investigated as a functional gene probe for detection of MSA-utilizing bacteria. PCR primers spanning a region of
msmA
which encoded a unique Rieske [2Fe-2S] binding region were designed. These primers were used to amplify the corresponding
msmA
genes from newly isolated
Hyphomicrobium, Methylobacterium
, and
Pedomicrobium
species that utilized MSA, from MSA enrichment cultures, and from DNA samples extracted directly from the environment. The high degree of identity of these
msmA
gene fragments, compared to
msmA
sequences from extant MSA utilizers, indicated the effectiveness of these PCR primers in molecular microbial ecology.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
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