Microbial Ecology

, Volume 71, Issue 4, pp 901–911 | Cite as

Pulsed 13C2-Acetate Protein-SIP Unveils Epsilonproteobacteria as Dominant Acetate Utilizers in a Sulfate-Reducing Microbial Community Mineralizing Benzene

  • Robert Starke
  • Andreas Keller
  • Nico Jehmlich
  • Carsten Vogt
  • Hans H. Richnow
  • Sabine Kleinsteuber
  • Martin von Bergen
  • Jana SeifertEmail author
Environmental Microbiology


In a benzene-degrading and sulfate-reducing syntrophic consortium, a clostridium affiliated to the genus Pelotomaculum was previously described to ferment benzene while various sulfate-reducing Deltaproteobacteria and a member of the Epsilonproteobacteria were supposed to utilize acetate and hydrogen as key metabolites derived from benzene fermentation. However, the acetate utilization network within this community was not yet unveiled. In this study, we performed a pulsed 13C2-acetate protein stable isotope probing (protein-SIP) approach continuously spiking low amounts of acetate (10 μM per day) in addition to the ongoing mineralization of unlabeled benzene. Metaproteomics revealed high abundances of Clostridiales followed by Syntrophobacterales, Desulfobacterales, Desulfuromonadales, Desulfovibrionales, Archaeoglobales, and Campylobacterales. Pulsed acetate protein-SIP results indicated that members of the Campylobacterales, the Syntrophobacterales, the Archaeoglobales, the Clostridiales, and the Desulfobacterales were linked to acetate utilization in descending abundance. The Campylobacterales revealed the fastest and highest 13C incorporation. Previous experiments suggested that the activity of the Campylobacterales was not essential for anaerobic benzene degradation in the investigated community. However, these organisms were consistently detected in various hydrocarbon-degrading and sulfate-reducing consortia enriched from the same aquifer. Here, we demonstrate that this member of the Campylobacterales is the dominant acetate utilizer in the benzene-degrading microbial consortium.


Anaerobic benzene degradation Metaproteomics Carbon flow 



We acknowledge the financial support by the German Research Foundation, Priority Program 1319. We are grateful to Michaela Wunderlich and Sibylle Mothes from the UFZ Department Analytical Chemistry for acetate analysis as well as Jörg Ahlheim and Werner Kletzander from the UFZ Department Groundwater Remediation for assistance in sampling of biological material from the columns. Benjamin Scheer is acknowledged for the support in using the Orbitrap in the ProVIS laboratory. The authors are grateful for using the analytical facilities of the Centre for Chemical Microscopy (ProVIS) at the Helmholtz Centre for Environmental Research which is supported by the European Regional Development Funds (EFRE–Europe funds Saxony) and the Helmholtz Association. We thank LABGeM and the National Infrastructure France Génomique for enabling the use of the annotation platform MicroScope.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Institute for Animal ScienceUniversity of HohenheimStuttgartGermany
  2. 2.Department of ProteomicsHelmholtz-Centre for Environmental Research (UFZ)LeipzigGermany
  3. 3.Department of Isotope BiogeochemistryHelmholtz-Centre for Environmental Research (UFZ)LeipzigGermany
  4. 4.Department of Environmental MicrobiologyHelmholtz-Centre for Environmental Research (UFZ)LeipzigGermany
  5. 5.Department of MetabolomicsHelmholtz-Centre for Environmental Research (UFZ)LeipzigGermany

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