Abstract
An anaerobic methylotrophic methanogenic enrichment culture, with sustained metabolic characteristics, including that of methanation for over a decade, was the choice of the present study on interspecies interactions. Growth and methanation by the enrichment were suppressed in the presence of antibiotics, and no methanogen grown on methanol could be isolated using stringent techniques. The present study confirmed syntrophic metabolic interactions in this enrichment with the isolation of a strain ofPseudomonas sp. The organism had characteristic metabolic versatility in metabolizing a variety of substrates including alcohols, aliphatic acids, amino acids, and sugars. Anaerobic growth was favoured with nitrate in the growth medium. Cells grown anaerobically with methanol, revealed maximal nitrate reductase activity. Constitutive oxidative activity of the membrane system emerged from the high-specific oxygen uptake and nitrate reductase activities of the aerobically and anerobically grown cells respectively. Cells grown anaerobically on various alcohols effectively oxidized methanol in the presence of flavins, cofactor FAD and the methanogenic cofactor F420, suggesting a constitutive alcohol oxidizing capacity. In cells grown anaerobically on methanol, the rate of methanol oxidation with F420 was three times that of FAD. Efficient utilization of alcohols in the presence of F420 is a novel feature of the present study. The results suggest that utilization of methanol by the mixed culture would involve metabolic interactions between thePseudomonas sp. and the methanogen(s). Methylotrophic, methanogenic partnership involving an aerobe is a novel feature hitherto unreported among anaerobic syntrophic associations and is of ecological significance.
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Albrecht R M, Rasmussen D H, Keller C S and Hinsdill R D 1976 Preparation of cultured cells for SEM: air drying from organic solvents;J. Microsc. 108 21–29
Anthony C and Zatman L J 1967 The microbial oxidation of methanol;Biochem. J. 104 960–969
Archer D B 1984 Biochemistry of methanogenesis by mixed cultures;Biochem. Soc. Trans. 12 1144–1146
Atlas R M 1997Handbook of microbiological media (ed.) L C Parks, 2nd edition (Boca Raton: CRC press)
Blaut M 1994 Metabolism of methanogens;Antonie Van Leeuwenhoek 66 187–208
Boone D R, Whitman W B and Rouvierie P 1993 Diversity and taxonomy of methanogens; inMethanogenesis (ed.) J G Ferry (New York, London: Chapman and Hall) pp 35–40
Bryant M P, Wolin E A, Wolin M J and Wolfe R S 1967Methanobacillus omelianskii, a symbiotic association of two species;Arch. Microbiol. 59 20–31
Bryant M P, McBride B C and Wolfe R S 1968 Hydrogen oxidizing methane bacteria. I. Cultivation and methanogenesis;J.Bacteriol. 95 1118–1123
Bryant M P, Campbell L L, Reddy C A and Crabill M R 1977 Growth ofDesulforvibrio in lactate or ethanol media low in sulphate in association with H2 using methanogenic bacteria;Appl. Environ. Microbiol. 33 1162–1169
Cole J 1996 Nitrate reduction to ammonia by enteric bacteria: redundancy, or a strategy for survival during oxygen starvation?;FEMS Microbiol. Lett. 136 1–11
Deppenmeier U, Blaut M, Mahlmann A and Gottschalk G 1990 Membrane-bound F420H2-dependent heterodisulfide reductase in methanogenic bacterium strain Go 1 andMethanolobus tindarius;FEBS Lett. 261 199–203
Fernandez-Lopez M, Olivares J and Bedmar E J 1994 Two differentially regulated nitrate reductases required for nitrate-dependent, microaerobic growth ofBradyrhizobium japonicum;Arch. Microbiol. 162 310–315
Hrywna Y, Tsoi T V, Maltseva O V, Quensen J F III and Tiedje J M 1999 Construction and characterization of two recombinant bacteria that grow on ortho- and para-substituted chlorobiphenyls;Appl. Environ. Microbiol. 65 2163–2169
Hubert C, Shen Y and Voordouw G 1999 Composition of toluenedegrading microbial communities from soil at different concentrations of toluene;Appl. Environ. Microbiol. 65 3064–3070
Hungate R E 1969 A roll tube method for cultivation of strict anaerobes; inMethods in microbiology (eds) J R Norris and D W Ribbons (New York: Academic Press) 3B, pp 117–132
King E O, Ward M K and Raney DE 1954 Two simple media for the demonstration of pyocyanin and the fluorescein;J. Lab. Clin. M ed. 44 301–307
Krishnan S and Lalitha K 1990 Interactive metabolic regulations during biomethanation ofLeucaena leucocepha;Appl. Biochem. Biotechnol. 26 73–85
Lalitha K, Swaminathan K R and Bai R P 1994a Kinetics of biomethanation of solid tannery waste and the concept of interactive metabolic control;Appl. Biochem. Biotechnol. 47 73–87
Lalitha K, Swaminathan K R, Vargheese C M, Shanthi V P and Bai R P 1994b Methanogenesis mediated by methylotrophic mixed culture;Appl. Biochem. Biotechnol. 49 113–134
Lange M and Ahring B K 2001 A comprehensive study into the molecular methodology and molecular biology of methanogenic Archaea;FEMS Microbiol. Rev. 25 553–571
Lo H and Reeves R E 1980 Purification and properties of NADPH:flavin oxidoreductase fromEntamoeba histolytica;Mol. Biochem. Parasitol. 2 23–30
McInerney M J 1988 Anaerobic degradation of proteins and lipids; inBiology of anaerobic microorganisms (ed.) A J B Zehnder (New York: John Wiley) pp 373–415
Meyer J M and Abdallah M A 1978 The fluorescent pigment ofPseudomonas fluorescens: Biosynthesis, purification and physicochemical properties;J. Gen. Microb. 107 319–328
Nicholas D J D and Nason A 1957 Determination of nitrate and nitrite;Methods Enzymol. 3 981–984
Palleroni N J 1984Pseudomonadaceae; inBergey’s manual of determinative bacteriology (eds) N R Krieg and J G Holt (Baltimore: Williams and Wilkins) Vol. 1, pp 141–198
Reid M F and Fewson C A 1994 Molecular characterization of microbial alcohol dehydrogenases;Crit. Rev. Microbiol. 20 13–56
Schink B 1997 Energetics of syntrophic cooperation in methanogenic degradation;Microbiol. Mol. Biol. Rev. 61 262–280
Sparling R and Gottschalk G 1990 Molecular hydrogen and energy conservation in methanogenic and acetogenic bacteria; inMicrobiology and biochemistry of strict anaerobes involved in interspecies hydrogen transfer (eds) J P Balaich, M Bruschi and J I Garcia (New York: Plenum Press) pp 3–10
Stams A J 1994 Metabolic interactions between anaerobic bacteria in methanogenic environments;Antonie Van Leeuwenhoek 66 271–294
Swaminathan K R 1995The concept of interactive metabolic control operative during biomethanation of solid tannery waste and methanol, Ph D thesis, Indian Institute of Technology Madras, Chennai, India
Thauer R K 1990 Energy metabolism of methanogenic bacteria;Biochim. Biophys. Acta 1018 256–259
Thauer R K 1998 Biochemistry of methanogenesis: a tribute to Marjory Stephenson;Microbiology 144 2377–2406
Thiele J H and Zeikus J G 1988 Control of interspecies electron flow during anaerobic digestion; Significance of formate transfer Vs hydrogen transfer during syntrophic methanogenesis in flocs;Appl. Environ. Microbiol. 54 20–29
Vasanthy N, Sankar K, Chandrasekaran P M and Lalitha K 1986 Biomethanation ofLeucaena leucocephala: a potential biomass substrate;Fue 165 1129–1133
Wolfaardt G M, Lawrence J R, Robarts R D and Caldwell D E 1994 The role of interactions, sessile growth, and nutrient amendments on the degradative efficiency of a microbial consortium;Can. J. Microbiol. 40 331–340
Wolin M J 1982 Hydrogen transfer in microbial communities; inMicrobial interactions and communities (eds) A T Bull and J H Slater (London: Academic Press) Vol. 1, pp 323–356
Zehnder A J B, Ingvorsen K and Marti T 1982 Microbiology of methane bacteria; inAnaerobic digestion (eds) D E Hughes, D A Stafford, B I Wheatley, W Baader, G Lettinga, E J Nyns, W Verstraete and R L Wentworth (Amsterdam: Elsevier Biomedical Press) pp 45–68
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Rapheal, S.V., Swaminathan, K.R. & Lalitha, K. Metabolic characteristics of an aerobe isolated from a methylotrophic methanogenic enrichment culture. J Biosci 28, 235–242 (2003). https://doi.org/10.1007/BF02706223
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DOI: https://doi.org/10.1007/BF02706223