Abstract
Dehalococcoides ethenogenes strain 195 (DE195) was grown in a sustainable syntrophic association with Desulfovibrio vulgaris Hildenborough (DVH) as a co-culture, as well as with DVH and the hydrogenotrophic methanogen Methanobacterium congolense (MC) as a tri-culture using lactate as the sole energy and carbon source. In the co- and tri-cultures, maximum dechlorination rates of DE195 were enhanced by approximately three times (11.0±0.01 μmol per day for the co-culture and 10.1±0.3 μmol per day for the tri-culture) compared with DE195 grown alone (3.8±0.1 μmol per day). Cell yield of DE195 was enhanced in the co-culture (9.0±0.5 × 107 cells per μmol Cl− released, compared with 6.8±0.9 × 107 cells per μmol Cl− released for the pure culture), whereas no further enhancement was observed in the tri-culture (7.3±1.8 × 107 cells per μmol Cl− released). The transcriptome of DE195 grown in the co-culture was analyzed using a whole-genome microarray targeting DE195, which detected 102 significantly up- or down-regulated genes compared with DE195 grown in isolation, whereas no significant transcriptomic difference was observed between co- and tri-cultures. Proteomic analysis showed that 120 proteins were differentially expressed in the co-culture compared with DE195 grown in isolation. Physiological, transcriptomic and proteomic results indicate that the robust growth of DE195 in co- and tri-cultures is because of the advantages associated with the capabilities of DVH to ferment lactate to provide H2 and acetate for growth, along with potential benefits from proton translocation, cobalamin-salvaging and amino acid biosynthesis, whereas MC in the tri-culture provided no significant additional benefits beyond those of DVH.
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References
Beaty PS, McInerney MJ . (1989). Effects of organic-acid anions on the growth and metabolism of Syntrophomonas wolfei in pure culture and in defined consortia. Appl Environ Microbiol 55: 977–983.
Benjamini Y, Hochberg Y . (1995). Controlling the false discovery rate: a practical and powerful approach to multiple testing. JR Stat Soc Ser B 57: 289–300.
Booker RS, Pavlostathis SG . (2000). Microbial redulctive dechlorination of hexachloro-1, 3-butadiene in a methanogenic enrichment culture. Water Res 34: 4437–4445.
Bryant MP, Campbell LL, Reddy CA, Crabill MR . (1977). Growth of Desulfovibrio in lactate or ethanol media low in sulfate in association with H2-utilizing methanogenic bacteria. Appl Environ Microbiol 33: 1162–1169.
Chourey K, Thompson MR, Morrel-Falvey J, VerBerkmoes NC, Brown SD, Shah M et al. (2006). Global molecular and morphological effects of 24-hour chromium(VI) exposure on Shewanella oneidensis MR-1. Appl Environ Microbiol 72: 6331–6344.
Cupples AM, Sporeman AM, McCarty PL . (2003). Growth of a Dehalococcoides-like microorganism on vinyl chloride and cis-dichloroethene as electron acceptors as determined by competitive PCR. Appl Environ Microbiol 69: 953–959.
DiStefano TD, Gossett JM, Zinder SH . (1992). Hydrogen as an electron donor for dechlorination of tetrachloroethene by an anaerobic mixed culture. Appl Environ Microbiol 58: 3622–3629.
Drzyzga O, Gottschal JC . (2002). Tetrachloroethene dehalorespiration and growth of Desulfitobactetium frappieri TCE1 in strict dependence on the activity of Desulfovibrio fructosivorans. Appl Environ Microbiol 68: 642–649.
Drzyzga O, Gerritse J, Dijk JA, Elissen H, Gottschal JC . (2001). Coexistence of a sulphate-reducing Desulfovibrio species and the dehalorespiring Deulfitobacterium frappieri TCE1 in defined chemostat cultures grown with various combinations of sulphate and tetrachloroethene. Environ Microbiol 3: 92–99.
Duhamel M, Edwards EA . (2006). Microbial composition of chlorinated ethane-degrading cultures dominated by Dehalococcoides. FEMS Microbiol Ecol 58: 538–549.
Eng JK, McCormack AL, Yates III JR . (1994). An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database. J Am Mass Spectrom 5: 976–989.
Fathepure BZ, Boyd SA . (1988). Reductive dechlorination of perchloroethylene and the role of methanogens. FEMS Microbiol Lett 49: 149–156.
Fennell DE, Gossett JM, Zinder SH . (1997). Comparison of butyric acid, ethanol, lactic acid, and propionic acid as hydrogen donors for the reductive dechlorination of tetrachloroethene. Environ Sci Technol 31: 918–926.
Freedman DL, Gossett JM . (1989). Biological reductive dechlorination of tetrachloroethylene and trichloroethylene to ethylene under methanogenic conditions. Appl Environ Microbiol 55: 2144–2151.
Gantzer CJ, Wackett LP . (1991). Reductive dechlorination catalyzed by bacterial transition-metal coenzymes. Environ Sci Technol 25: 715–722.
Gentleman RC, Carey VJ, Bates DM, Bolstad B, Dettling M, Dudoit S et al. (2004). Bioconductor: open software development for computational biology and bioinformatics. Genome Biol 5: R80.
He J, Holmes V, Lee PKH, Alvarez-Cohen L . (2007). Influence of vitamin B12 and co-cultures on the growth of Dehalococcoides isolates in defined medium. Appl Environ Microbiol 73: 2847–2853.
He J, Ritalahti KM, Aiello MR, Löffler FE . (2003a). Complete detoxification of vinyl chloride (VC) by an anaerobic enrichment culture and identification of the reductively dechlorinating population as a Dehalococcoides species. Appl Environ Microbiol 69: 996–1003.
He J, Ritalahti KM, Yang KL, Koenigsberg SS, Löffler FE . (2003b). Detoxification of vinyl chloride to ethene coupled to growth of an anaerobic bacterium. Nature 424: 62–65.
Heimann AC, Batstone DJ, Jakobsen R . (2006). Methanosarcina spp. drive vinyl chloride dechlorination via interspecies hydrogen transfer. Appl Environ Microbiol 72: 2942–2949.
Holliger C, Wohlfarth G, Diekert G . (1999). Reductive dechlorination in the energy metabolism of anaerobic bacteria. FEMS Microbiol Rev 22: 383–398.
Horwitz W (ed.) (2000). Official Methods of Analysis of AOAC International, 17th edn. AOAC International: Gaithersburg, MD.
Johnson DR, Brodie EL, Hubbard AE, Andersen GL, Zinder SH, Alvarez-Cohen L . (2008). Temporal transcriptomic microarray analysis of Dehalococcoides ethenogenes strain 195 during the transition from exponential growth to the stationary phase. Appl Environ Microbiol 74: 2864–2872.
Johnson DR, Nemir A, Anderson GL, Zinder SH, Alvarez-Cohen L . (2009). Transcriptomic microarray analysis of corrinoid responsive genes in Dehalococcoides ethenogenes strain 195. FEMS Microbiol Lett 294: 198–206.
Lee PKH, Johnson DR, Holmes VF, He J, Alvarez-Cohen L . (2006). Reductive dehalogenase gene expression as a biomarker for physiological activity of Dehalococcoides spp. Appl Environ Microbiol 72: 6161–6168.
Löffler FE, Ritalahti KM, Tiedje JM . (1997). Dechlorination of chloroethenes is inhibited by 2-bromoethanesulfonate in the absence of methanogens. Appl Environ Microbiol 63: 4982–4985.
Maymó-Gatell X, Chien YT, Gossett JM, Zinder SH . (1997). Isolation of a bacterium that reductively dechlorinates tetrachloroethene to ethene. Science 276: 1568–1571.
McINerney MJ, Bryant MP . (1981). Anaerobic degredation of lactate by syntrophic associations of Methanosarcina barkeri and Desulfovibrio species and effect of H2 on acetate degredation. Appl Environ Microbiol 41: 346–354.
Ramos JL, MartÃnez-Bueno M, Molina-Henares AJ, Terán W, Watanabe K, Zhang X et al. (2005). The TetR family of transcriptional repressors. Microbiol Mol Biol Rev 69: 326–356.
Richardson RE, Bhupathiraju VK, Song DL, Goulet TA, Alvarez-Cohen L . (2002). Phylogenetic characterization of microbial communities that reductively dechlorinate TCE based upon a combination of molecular techniques. Environ Sci Technol 36: 2652–2662.
Ritalahti KM, Amos BK, Sung Y, Wu Q, Koenigsberg SS, Löffler FE . (2006). Quantitative PCR targeting 16S rRNA and reductive dehalogenase genes simultaneously monitors multiple Dehalococcoides strains. Appl Environ Microbiol 72: 2765–2774.
Ritalahti KM, Löffler FE . (2004). Populations implicated in the anaerobic reductive dechlorination of 1, 2-dichloropropane in highly enriched bacterial communities. Appl Environ Microbiol 70: 4088–4095.
Rittmann BE, McCarty PL . (2001). Environmental Biotechnology: Principles and Applications. McGraw-Hill: Boston, pp 570–629.
Rodionov DA, Dubchak I, Arkin A, Alm E, Gelfand MS . (2004). Reconstruction of regulatory and metabolic pathways in metal-reducing ä-proteobacteria. Genome Biol 5: R90.
Schink B . (1997). Energetics of syntrophic cooperation in methanogenic degradation. Microbiol Mol Biol Rev 61: 262–280.
Scholten JC, Culley DE, Brockman FJ, Wu G, Zhang WW . (2007). Evolution of the syntrophic interaction between Desulfovibrio vulgaris and Methanosarcina barkeri: involvement of an ancient horizontal gene transfer. Biochem Biophys Res Commun 352: 48–54.
Seshadri R, Adrian L, Fouts DE, Eisen JA, Phillippy AM, Methe BA et al. (2005). Genome sequence of the PCE-dechlorinating bacterium Dehalococcoides ethenogenes. Science 307: 105–108.
Smatlak CR, Gossett JM, Zinder SH . (1996). Comparative kinetics of hydrogen utilization for reductive dechlorination of tetrachloroethene and methanogenesis in an anaerobic enrichment culture. Environ Sci Technol 30: 2850–2858.
Smidt H, de Vos WM . (2004). Anaerobic microbial dehalogenation. Annu Rev Microbiol 58: 43–73.
Sofia HJ, Chen G, Hetzler BG, Reyes-Spindola JF, Miller NE . (2001). Radical SAM, a novel protein superfamily linking unresolved steps in familiar biosynthetic pathways with radical mechanisms: functional characterization using new analysis and information visualization methods. Nucleic Acids Res 29: 1097–1106.
Stolyar S, Van Dien S, Hillesland KL, Lie TJ, Leigh JA, Stahl DA . (2007). Metabolic modeling of a mutualistic microbial community. Mol Syst Biol 3: 92.
Tabb DL, McDonald WH, Yates III JR . (2002). DTASelect and contrast: tools for assembling and comparing protein identifications from shotgun proteomics. J Proteome Res 1: 21–26.
Tropel D, van der Meer JR . (2004). Bacterial transcriptional regulators for degradation pathways of aromatic compounds. Microbiol Mol Biol Rev 68: 474–500.
Verberkmoes NC, Russell AL, Shah M, Godzik A, Rosenquist M, Halfvarson J et al. (2009). Shotgun metaproteomics of the human distal gut microbiota. ISME J 3: 179–189.
Vogel TM, McCarty PL . (1985). Biotransformation of tetrachloroethylene to trichloroethylene, dichloroethylene, vinyl-chloride, and carbon-dioxide under methanogenic conditions. Appl Environ Microbiol 49: 1080–1083.
Walker CB, He ZL, Yang ZK, Ringbauer JA, He Q, Zhou JH et al. (2009). The electron transfer system of syntrophically grown Desulfovibrio vulgaris. J Bacteriol 191: 5793–5801.
West KA, Johnson DR, Hu P, DeSantis TZ, Brodie EL, Lee PKH et al. (2008). Comparative genomics of Dehalococcoides ethenogenes 195 and a Dehalococcoides-containing enrichment culture. Appl Environ Microbiol 74: 3533–3540.
Yang YR, McCarty PL . (1998). Competition for hydrogen within a chlorinated solvent dehalogenating anaerobic mixed culture. Environ Sci Technol 32: 3591–3597.
Yi S, Zhuang WQ, Feng X, Zinder SH, Tang YJ, Alvarez-Cohen L . (2010). Exogenous amino acid utilization by Dehalococcoides ethenogenes strain 195. 110th General Meeting of the American Society for Microbiology, San Diego, CA.
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Men, Y., Feil, H., VerBerkmoes, N. et al. Sustainable syntrophic growth of Dehalococcoides ethenogenes strain 195 with Desulfovibrio vulgaris Hildenborough and Methanobacterium congolense: global transcriptomic and proteomic analyses. ISME J 6, 410–421 (2012). https://doi.org/10.1038/ismej.2011.111
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DOI: https://doi.org/10.1038/ismej.2011.111
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