Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 106, Issue 8, Pages 2874-2879Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0806798106
Keywords
central carbon metabolism; genome context analysis; lactate dehydrogenase
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Funding
- U. S. Department of Energy (DOE) Office of Biological and Environmental Research under the Genomics
- DOE by Battelle Memorial Institute [DE-AC05-76RLO 1830.]
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The ability to use lactate as a sole source of carbon and energy is one of the key metabolic signatures of Shewanellae, a diverse group of dissimilatory metal-reducing bacteria commonly found in aquatic and sedimentary environments. Nonetheless, homology searches failed to recognize orthologs of previously described bacterial D- or L-lactate oxidizing enzymes (Escherichia coli genes dId and IIdD) in any of the 13 analyzed genomes of Shewanella spp. By using comparative genomic techniques, we identified a conserved chromosomal gene cluster in Shewanella oneidensis MR-1 (locus tag: SO_1522-SO_1518) containing lactate permease and candidate genes for both D- and L-lactate dehydrogenase enzymes. The predicted D-LDH gene (dId-II, SO_1521) is a distant homolog of FAD-dependent lactate dehydrogenase from yeast, whereas the predicted L-LDH is encoded by 3 genes with previously unknown functions (IIdEGF, SO_1520-SO_1518). Through a combination of genetic and biochemical techniques, we experimentally confirmed the predicted physiological role of these novel genes in S. oneidensis MR-1 and carried out successful functional validation studies in Escherichia coli and Bacillus subtilis. We conclusively showed that dId-II and IIdEFG encode fully functional D- and L-LDH enzymes, which catalyze the oxidation of the respective lactate stereoisomers to pyruvate. Notably, the S. oneidensis MR-1 LIdEFG enzyme is a previously uncharacterized example of a multisubunit lactate oxidase. Comparative analysis of > 400 bacterial species revealed the presence of LIdEFG and DId-II in a broad range of diverse species accentuating the potential importance of these previously unknown proteins in microbial metabolism.
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