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Journal of Bacteriology, December 1998, p. 6292-6297, Vol. 180, No. 23
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Shewanella putrefaciens mtrB Encodes an
Outer Membrane Protein Required for Fe(III) and Mn(IV)
Reduction
Alexander S.
Beliaev and
Daad A.
Saffarini*
Department of Microbiology, University of
Massachusetts, Amherst, Massachusetts 01003
Received 13 May 1998/Accepted 30 September 1998
Iron and manganese oxides or oxyhydroxides are abundant transition
metals, and in aquatic environments they serve as terminal electron
acceptors for a large number of bacterial species. The molecular
mechanisms of anaerobic metal reduction, however, are not understood.
Shewanella putrefaciens is a facultative anaerobe that uses Fe(III) and Mn(IV) as terminal electron acceptors
during anaerobic respiration. Transposon mutagenesis was used to
generate mutants of S. putrefaciens, and one such mutant,
SR-21, was analyzed in detail. Growth and enzyme assays indicated that
the mutation in SR-21 resulted in loss of Fe(III) and Mn(IV) reduction
but did not affect its ability to reduce other electron acceptors used
by the wild type. This deficiency was due to Tn5
inactivation of an open reading frame (ORF) designated mtrB.
mtrB encodes a protein of 679 amino acids and contains a signal
sequence characteristic of secreted proteins. Analysis of membrane
fractions of the mutant, SR-21, and wild-type cells indicated that MtrB
is located on the outer membrane of S. putrefaciens. A
5.2-kb DNA fragment that contains mtrB was isolated and
completely sequenced. A second ORF, designated mtrA, was
found directly upstream of mtrB. The two ORFs appear to be
arranged in an operon. mtrA encodes a putative 10-heme
c-type cytochrome of 333 amino acids. The N-terminal
sequence of MtrA contains a potential signal sequence for secretion
across the cell membrane. The amino acid sequence of MtrA exhibited
34% identity to NrfB from Escherichia coli, which is
involved in formate-dependent nitrite reduction. To our knowledge, this
is the first report of genes encoding proteins involved in metal reduction.
*
Corresponding author. Present address: University of
Wisconsin
Milwaukee, Department of Biological Sciences, 3209 N. Maryland Ave., Milwaukee, WI 53211. Phone: (414) 229-2964. Fax: (414)
229-3926. E-mail: daads{at}uwm.edu.
Journal of Bacteriology, December 1998, p. 6292-6297, Vol. 180, No. 23
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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