Applied Microbiology and Biotechnology

, Volume 85, Issue 2, pp 389–403 | Cite as

Effect of nutrient and selective inhibitor amendments on methane oxidation, nitrous oxide production, and key gene presence and expression in landfill cover soils: characterization of the role of methanotrophs, nitrifiers, and denitrifiers

  • Sung-Woo Lee
  • Jeongdae Im
  • Alan A. DiSpirito
  • Levente Bodrossy
  • Michael J. Barcelona
  • Jeremy D. SemrauEmail author
Environmental Biotechnology


Methane and nitrous oxide are both potent greenhouse gasses, with global warming potentials approximately 25 and 298 times that of carbon dioxide. A matrix of soil microcosms was constructed with landfill cover soils collected from the King Highway Landfill in Kalamazoo, Michigan and exposed to geochemical parameters known to affect methane consumption by methanotrophs while also examining their impact on biogenic nitrous oxide production. It was found that relatively dry soils (5% moisture content) along with 15 mg NH 4 + (kg soil)−1 and 0.1 mg phenylacetylene∙(kg soil)−1 provided the greatest stimulation of methane oxidation while minimizing nitrous oxide production. Microarray analyses of pmoA showed that the methanotrophic community structure was dominated by Type II organisms, but Type I genera were more evident with the addition of ammonia. When phenylacetylene was added in conjunction with ammonia, the methanotrophic community structure was more similar to that observed in the presence of no amendments. PCR analyses showed the presence of amoA from both ammonia-oxidizing bacteria and archaea, and that the presence of key genes associated with these cells was reduced with the addition of phenylacetylene. Messenger RNA analyses found transcripts of pmoA, but not of mmoX, nirK, norB, or amoA from either ammonia-oxidizing bacteria or archaea. Pure culture analyses showed that methanotrophs could produce significant amounts of nitrous oxide, particularly when expressing the particulate methane monooxygenase (pMMO). Collectively, these data suggest that methanotrophs expressing pMMO played a role in nitrous oxide production in these microcosms.


Methanotroph Landfill Nitrous oxide Methane Ammonia oxidizers 



We thank George Wells and Craig Criddle for providing an amoA amplicon from ammonia-oxidizing archaea. Support from the Department of Energy (DE-FC26-05NT42431) to JDS is also gratefully acknowledged.


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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Sung-Woo Lee
    • 1
    • 5
  • Jeongdae Im
    • 1
  • Alan A. DiSpirito
    • 2
  • Levente Bodrossy
    • 3
  • Michael J. Barcelona
    • 4
  • Jeremy D. Semrau
    • 1
    Email author
  1. 1.Department of Civil and Environmental EngineeringThe University of MichiganAnn ArborUSA
  2. 2.Department of Biochemistry, Biophysics and Molecular BiologyIowa State UniversityAmesUSA
  3. 3.Department of BiotechnologyARC Seibersdorf Research GmbHSeibersdorfAustria
  4. 4.Department of ChemistryWestern Michigan UniversityKalamazooUSA
  5. 5.Oregon Graduate Institute, Department of Environmental and Biomolecular SystemsOregon Health and Sciences UniversityBeavertonUSA

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