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Copper enhances the activity and salt resistance of mixed methane-oxidizing communities

  • ENVIRONMENTAL BIOTECHNOLOGY
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Abstract

Effluents of anaerobic digesters are an underestimated source of greenhouse gases, as they are often saturated with methane. A post-treatment with methane-oxidizing bacterial consortia could mitigate diffuse emissions at such sites. Semi-continuously fed stirred reactors were used as model systems to characterize the influence of the key parameters on the activity of these mixed methanotrophic communities. The addition of 140 mg L−1 NH +4 –N had no significant influence on the activity nor did a temperature increase from 28°C to 35°C. On the other hand, addition of 0.64 mg L−1 of copper(II) increased the methane removal rate by a factor of 1.5 to 1.7 since the activity of particulate methane monooxygenase was enhanced. The influence of different concentrations of NaCl was also tested, as effluents of anaerobic digesters often contain salt levels up to 10 g NaCl L−1. At a concentration of 11 g NaCl L−1, almost no methane-oxidizing activity was observed in the reactors without copper addition. Yet, reactors with copper addition exhibited a sustained activity in the presence of NaCl. A colorimetric test based on naphthalene oxidation showed that soluble methane monooxygenase was inhibited by copper, suggesting that the particulate methane monooxygenase was the active enzyme and thus more salt resistant. The results obtained demonstrate that the treatment of methane-saturated effluents, even those with increased ammonium (up to 140 mg L−1 NH +4 –N) and salt levels, can be mitigated by implementation of methane-oxidizing microbial consortia.

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References

  • Amaral JA, Knowles R (1995) Growth of methanotrophs in methane and oxygen counter gradients. FEMS Microbiol Lett 126:215–220

    Article  CAS  Google Scholar 

  • Anthony C (1982) Biochemistry of methylotrophs. Academic, London

    Google Scholar 

  • Balasubramanian R, Rosenzweig AC (2008) Copper methanobactin: a molecule whose time has come. Curr Opin Chem Biol 12:245–249

    Article  CAS  Google Scholar 

  • Begonja A, Hrsak D (2001) Effect of growth conditions on the expression of soluble methane monooxygenase. Food Technol Biotechnol 39:29–35

    CAS  Google Scholar 

  • Bender M, Conrad R (1992) Kinetics of CH4 oxidation in oxic soils exposed to ambient air or high CH4 mixing ratios. FEMS Microbiol Ecol 101:261–270

    Article  CAS  Google Scholar 

  • Bender M, Conrad R (1995) Effect of CH4 concentrations and soil conditions on the induction of CH4 oxidation activity. Soil Biol Biochem 27:1517–1527

    Article  CAS  Google Scholar 

  • Bodelier PLE, Laanbroek HJ (2004) Nitrogen as a regulatory factor of methane oxidation in soils and sediments. FEMS Microbiol Ecol 47:265–277

    Article  CAS  Google Scholar 

  • Bodelier PLE, Meima-Franke M, Zwart G, Laanbroek HJ (2005) New DGGE strategies for the analyses of methanotrophic microbial communities using different combinations of existing 16S rRNA-based primers. FEMS Microbiol Ecol 52:163–174

    Article  CAS  Google Scholar 

  • Boon N, Goris J, De Vos P, Verstraete W, Top EM (2000) Bioaugmentation of activated sludge by an indigenous 3-chloroaniline-degrading Comamonas testosteroni strain, I2gfp. Appl Environ Microbiol 66:2906–2913

    Article  CAS  Google Scholar 

  • Bowman JP, Sly LI, Nichols PD, Hayward AC (1993) Revised taxonomy of the methanotrophs—description of Methylobacter Gen-Nov, emendation of Methylococcus, validation of Methylosinus and Methylocystis species, and a proposal that the family Methylococcaceae includes only the Group-I methanotrophs. Int J Syst Bacteriol 43:735–753

    Article  Google Scholar 

  • Brusseau GA, Tsien HC, Hanson RS, Wackett LP (1990) Optimization of trichloroethylene oxidation by methanotrophs and the use of a colorimetric assay to detect soluble methane monooxygenase activity. Biodegradation 1:19–29

    Article  CAS  Google Scholar 

  • Bullock CM, Bicho PA, Zhang Y, Saddler JN (1996) A solid chemical oxygen demand (COD) method for determining biomass in waste waters. Water Res 30:1280–1284

    Article  CAS  Google Scholar 

  • Cakir FY, Stenstrom MK (2005) Greenhouse gas production: a comparison between aerobic and anaerobic wastewater treatment technology. Water Res 39:4197–4203

    Article  CAS  Google Scholar 

  • Colby J, Stirling DI, Dalton H (1977) Soluble methane mono-oxygenase of Methylococcus capsulatus (Bath)—ability to oxygenate normal-alkanes, normal-alkenes, ethers, and alicyclic, aromatic and heterocyclic compounds. Biochem J 165:395–402

    CAS  Google Scholar 

  • Csonka LN (1989) Physiological and gentic responses of bacteria to osmotic-stress. Microbiol Rev 53:121–147

    CAS  Google Scholar 

  • Dalton H (2005) The Leeuwenhoek Lecture 2000—the natural and unnatural history of methane-oxidizing bacteria. Philos Trans R Soc Lond, Ser B 360:1207–1222

    Article  CAS  Google Scholar 

  • De Visscher A, Van Cleemput O (2003) Induction of enhanced CH4 oxidation in soils: NH +4 inhibition patterns. Soil Biol Biochem 35:907–913

    Article  Google Scholar 

  • Dunfield PF, Liesack W, Henckel T, Knowles R, Conrad R (1999) High-affinity methane oxidation by a soil enrichment culture containing a type II methanotroph. Appl Environ Microbiol 65:1009–1014

    CAS  Google Scholar 

  • Gourdon R, Comel C, Vermande P, Veron J (1989) Kinetics of acetate, propionate and butyrate removal in the treatment of a semi-synthetic landfill leachate on anaerobic filter. Biotechnol Bioeng 33:1167–1181

    Article  CAS  Google Scholar 

  • Greenberg AE, Clesceri LS, Eaton AD (eds) (1992) Standard methods for the examination of water and wastewater, 18th edn. American Public Health Association, American Water Works Association, Water Environment Federation, Washington

    Google Scholar 

  • Gulledge J, Schimel JP (1998) Low-concentration kinetics of atmospheric CH4 oxidation in soil and mechanism of NH +4 inhibition. Appl Environ Microbiol 64:4291–4298

    CAS  Google Scholar 

  • Hakemian AS, Rosenzweig AC (2007) The biochemistry of methane oxidation. Annu Rev Biochem 76:223–241

    Article  CAS  Google Scholar 

  • Hanson RS, Hanson TE (1996) Methanotrophic bacteria. Microbiol Rev 60:439–471

    CAS  Google Scholar 

  • Hartley K, Lant P (2006) Eliminating non-renewable CO2 emissions from sewage treatment: An anaerobic migrating bed reactor pilot plant study. Biotechnol Bioeng 95:384–398

    Article  CAS  Google Scholar 

  • Helm J, Wendlandt KD, Rogge G, Kappelmeyer U (2006) Characterizing a stable methane-utilizing mixed culture used in the synthesis of a high-quality biopolymer in an open system. J Appl Microbiol 101:387–395

    Article  CAS  Google Scholar 

  • Henckel T, Jackel U, Schnell S, Conrad R (2000) Molecular analyses of novel methanotrophic communities in forest soil that oxidize atmospheric methane. Appl Environ Microbiol 66:1801–1808

    Article  CAS  Google Scholar 

  • Hrsak D, Begonja A (2000) Possible interactions within a methanotrophic–heterotrophic groundwater community able to transform linear alkylbenzenesulfonates. Appl Environ Microbiol 66:4433–4439

    Article  CAS  Google Scholar 

  • IPCC (2001) Climate change 2001: the scientific basis. In: Houghton J, Ding Y, Griggs D, van der Linden P (eds) Contribution of working group I to the third assesment report of the intergovernmental panel of climate change. Cambridge University Press, Cambridge

    Google Scholar 

  • Ismail SB, Gonzalez P, Jeison D, van Lier JB (2008) Effects of high salinity wastewater on methanogenic sludge bed systems. Water Sci Technol 58:1963–1970

    Article  CAS  Google Scholar 

  • Jahng D, Wood TK (1996) Metal ions and chloramphenicol inhibition of soluble methane monooxygenase from Methylosinus trichosporium OB3b. Appl Microbiol Biotechnol 45:744–749

    Article  CAS  Google Scholar 

  • Joergensen L, Degn H (1987) Growth-rate and methane affinity of a turbidostatic and oxystatic continuous culture of Methylococcus capsulatus (Bath). Biotechnol Lett 9:71–76

    Article  CAS  Google Scholar 

  • King GM, Schnell S (1998) Effects of ammonium and non-ammonium salt additions on methane oxidation by Methylosinus trichosporium OB3b and Maine forest soils. Appl Environ Microbiol 64:253–257

    CAS  Google Scholar 

  • Koh SC, Bowman JP, Sayler GS (1993) Soluble methane monooxygenase production and trichloroethylene degradation by a type-I methanotroph, Methylomons methanica-68-1. Appl Environ Microbiol 59:960–967

    CAS  Google Scholar 

  • Melse RW, Van der Werf AW (2005) Biofiltration for mitigation of methane emission from animal husbandry. Environ Sci Technol 39:5460–5468

    Article  CAS  Google Scholar 

  • Merkx M, Kopp DA, Sazinsky MH, Blazyk JL, Muller J, Lippard SJ (2001) Dioxygen activation and methane hydroxylation by soluble methane monooxygenase: a tale of two irons and three proteins. Angew Chem Int Ed 40:2782–2807

    Article  CAS  Google Scholar 

  • Nyerges G, Stein LY (2009) Ammonia cometabolism and product inhibition vary considerably among species of methanotrophic bacteria. FEMS Microbiol Lett 297:131–136

    Article  CAS  Google Scholar 

  • Park JR, Moon S, Ahn YM, Kim JY, Nam K (2005) Determination of environmental factors influencing methane oxidation in a sandy landfill cover soil. Environ Technol 26:93–102

    Article  CAS  Google Scholar 

  • Scheutz C, Kjeldsen P (2004) Environmental factors influencing attenuation of methane and hydrochlorofluorocarbons in landfill cover soils. J Environ Qual 33:72–79

    Article  CAS  Google Scholar 

  • Scheutz C, Kjeldsen P, Bogner JE, De Visscher A, Gebert J, Hilger HA, Huber-Humer M, Spokas K (2009) Microbial methane oxidation processes and technologies for mitigation of landfill gas emissions. Waste Manage Res 27:409–455

    Article  CAS  Google Scholar 

  • Schnell S, King GM (1996) Responses of methanotrophic activity in soils and cultures to water stress. Appl Environ Microbiol 62:3203–3209

    CAS  Google Scholar 

  • Shah NN, Park S, Taylor RT, Droege MW (1992) Cultivation of Methylosinus trichosporium Ob3b. 3. Production of particulate methane monooxygenase in continuous culture. Biotechnol Bioeng 40:705–712

    Article  CAS  Google Scholar 

  • Trotsenko YA, Khmelenina VN (2002) Biology of extremophilic and extremotolerant methanotrophs. Arch Microbiol 177:123–131

    Article  CAS  Google Scholar 

  • Whalen SC (2000) Influence of N and non-N salts on atmospheric methane oxidation by upland boreal forest and tundra soils. Biol Fertil Soils 31:279–287

    Article  CAS  Google Scholar 

  • Whittenbury R, Phillips KC, Wilkinson JF (1970) Enrichment, isolation and some properties of methane-utilizing bacteria. J Gen Microbiol 61:205–217

    CAS  Google Scholar 

  • Wilshusen JH, Hettiaratchi JPA, De Visscher A, Saint-Fort R (2004) Methane oxidation and formation of EPS in compost: effect of oxygen concentration. Environ Pollut 129:305–314

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by a Ph.D. grant (no. 83259) from the Institute for the Promotion of Innovation through Science and Technology in Flanders (IWT-Vlaanderen), a research grant from the Flemish Fund for Scientific Research (FWO-Vlaanderen, 3G070010), and by the Geconcerteerde Onderzoeksactie (GOA) of Ghent University (BOF09/GOA/005). Tim Lacoere’s help with the molecular analyses is greatly acknowledged. Also, thanks to Jan Vermeulen and Samuel Bodé for their support with GC analyses and Vicky D’havé for her assistance. Furthermore, Anthony Hay, Joachim Desloover, Yu Zhang, and Willem De Muynck are gratefully appreciated for critically reading the manuscript.

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Authors and Affiliations

  1. Laboratory of Microbial Ecology and Technology (LabMET), Ghent University, Coupure Links 653, 9000, Ghent, Belgium

    David van der Ha, Willy Verstraete & Nico Boon

  2. Laboratory of Microbiology (LM-UGent), Ghent University, K.L. Ledeganckstraat 35, 9000, Ghent, Belgium

    Sven Hoefman

  3. Laboratory of Applied Physical Chemistry (ISOFYS), Ghent University, Coupure Links 653, 9000, Ghent, Belgium

    Pascal Boeckx

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  1. David van der Ha
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  2. Sven Hoefman
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  4. Willy Verstraete
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  5. Nico Boon
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Correspondence to Nico Boon.

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van der Ha, D., Hoefman, S., Boeckx, P. et al. Copper enhances the activity and salt resistance of mixed methane-oxidizing communities. Appl Microbiol Biotechnol 87, 2355–2363 (2010). https://doi.org/10.1007/s00253-010-2702-4

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  • DOI: https://doi.org/10.1007/s00253-010-2702-4

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