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

  • Environmental Microbiology
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Abstract

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.

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Acknowledgments

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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Authors and Affiliations

  1. Institute for Animal Science, University of Hohenheim, Emil-Wolff-Str. 6-10, 70599, Stuttgart, Germany

    Robert Starke & Jana Seifert

  2. Department of Proteomics, Helmholtz-Centre for Environmental Research (UFZ), Permoserstr. 15, 04318, Leipzig, Germany

    Robert Starke, Nico Jehmlich & Martin von Bergen

  3. Department of Isotope Biogeochemistry, Helmholtz-Centre for Environmental Research (UFZ), Permoserstr. 15, 04318, Leipzig, Germany

    Andreas Keller, Carsten Vogt & Hans H. Richnow

  4. Department of Environmental Microbiology, Helmholtz-Centre for Environmental Research (UFZ), Permoserstr. 15, 04318, Leipzig, Germany

    Andreas Keller & Sabine Kleinsteuber

  5. Department of Metabolomics, Helmholtz-Centre for Environmental Research (UFZ), Permoserstr. 15, 04318, Leipzig, Germany

    Martin von Bergen

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  1. Robert Starke
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  2. Andreas Keller
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Starke, R., Keller, A., Jehmlich, N. et al. Pulsed 13C2-Acetate Protein-SIP Unveils Epsilonproteobacteria as Dominant Acetate Utilizers in a Sulfate-Reducing Microbial Community Mineralizing Benzene. Microb Ecol 71, 901–911 (2016). https://doi.org/10.1007/s00248-016-0731-y

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