Spatial separation of metabolic stages in a tube anaerobic baffled reactor: reactor performance and microbial community dynamics
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Spatial separation of metabolic stages in anaerobic digesters can increase the methane content of biogas, as realized in a tube anaerobic baffled reactor. Here, we investigated the performance and microbial community dynamics of a laboratory-scale mesophilic anaerobic baffled reactor with four compartments treating an artificial substrate. Due to the activity of fermentative bacteria, organic acids mostly accumulated in the initial compartments. The methane content of the biogas increased while hydrogen levels decreased along the compartments. Microbial communities were investigated based on bacterial 16S rRNA genes, hydA genes encoding Fe–Fe-hydrogenases, and mcrA genes/transcripts encoding the methyl-CoM reductase. The metaproteome was analyzed to identify active metabolic pathways. During the reactor operation, Clostridia and Bacilli became most abundant in the first compartment. Later compartments were dominated by Sphingobacteriia, Deltaproteobacteria, Clostridia, Bacteroidia, Synergistia, Anaerolineae, Spirochaetes, vadinHA17, and W5 classes. Methanogenic communities were represented by Methanomicrobiales, Methanobacteriaceae, Methanosaeta, and Methanosarcina in the last compartments. Analysis of hydA and mcrA genes and metaproteome data confirmed the spatial separation of metabolic stages. In the first compartment, proteins of carbohydrate transport and metabolism were most abundant. Proteins assigned to coenzyme metabolism and transport as well as energy conservation dominated in the other compartments. Our study demonstrates how the spatial separation of metabolic stages by reactor design is underpinned by the adaptation of the microbial community to different niches.
KeywordsAnaerobic digestion Biogas Amplicon pyrosequencing hydA mcrA Metaproteome
The authors wish to thank Ute Lohse for technical support in amplicon sequencing.
This study was funded by the Russian Foundation for Basic Research (Grant No. 16-34-60093 mol_a_dk and Grant No. 18-29-25058 awarded to AMZ).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Abdo Z, Schütte UM, Bent SJ, Williams CJ, Forney LJ, Joyce P (2006) Statistical methods for characterizing diversity of microbial communities by analysis of terminal restriction fragment length polymorphisms of 16S rRNA genes. Environ Microbiol 8:929–938. https://doi.org/10.1111/j.1462-2920.2005.00959.x CrossRefGoogle Scholar
- Cabezas A, Calabria de Araujo J, Callejas C, Galès A, Hamelin J, Marone A, Sousa DZ, Trably E, Etchebehere C (2015) How to use molecular biology tools for the study of the anaerobic digestion process? Rev Environ Sci Biotechnol 14:555–593. https://doi.org/10.1007/s11157-015-9380-8 CrossRefGoogle Scholar
- Chung KT (1976) Inhibitory effects of H2 on growth of Clostridium cellobioparum. Appl Environ Microbiol 31:342–348Google Scholar
- Haange SB, Oberbach A, Schlichting N, Hugenholtz F, Smidt H, von Bergen M, Till H, Seifert J (2012) Metaproteome analysis and molecular genetics of rat intestinal microbiota reveals section and localization resolved species distribution and enzymatic functionalities. J Proteome Res 11:5406–5417. https://doi.org/10.1021/pr3006364 CrossRefGoogle Scholar
- Heyer R, Benndorf D, Kohrs F, De Vrieze J, Boon N, Hoffmann M, Rapp E, Schlüter A, Sczyrba A, Reichl U (2016) Proteotyping of biogas plant microbiomes separates biogas plants according to process temperature and reactor type. Biotechnol Biofuels 9:155. https://doi.org/10.1186/s13068-016-0572-4 CrossRefGoogle Scholar
- McInerney MJ, Bryant MP, Hespell RB, Costerton JW (1981) Syntrophomonas wolfei gen. nov. sp. nov., an anaerobic, syntrophic, fatty acid-oxidizing bacterium. Appl Environ Microbiol 41:1029–1039Google Scholar
- Nikolausz M, Walter RF, Sträuber H, Liebetrau J, Schmidt T, Kleinsteuber S, Bratfisch F, Günther U, Richnow HH (2013) Evaluation of stable isotope fingerprinting techniques for the assessment of the predominant methanogenic pathways in anaerobic digesters. Appl Microbiol Biotechnol 97:2251–2262. https://doi.org/10.1007/s00253-012-4657-0 CrossRefGoogle Scholar
- Oksanen J (2011) Multivariate analysis of ecological communities in R: vegan tutorial. Publisher Univ Oulu Comput Serv Cent 83:1–43Google Scholar
- Oliveros JC (2007-2015) Venny. An interactive tool for comparing lists with Venn’s diagrams. http://bioinfogp.cnb.csic.es/tools/venny/index.html
- Pelletier E, Kreimeyer A, Bocs S, Rouy Z, Gyapay G, Chouari R, Rivière D, Ganesan A, Daegelen P, Sghir A, Cohen GN, Médigue C, Weissenbach J, Le Paslier D (2008) “Candidatus Cloacamonas acidaminovorans”: genome sequence reconstruction provides a first glimpse of a new bacterial division. J Bacteriol 190:2572–2579. https://doi.org/10.1128/JB.01248-07 CrossRefGoogle Scholar
- Purushe J, Fouts DE, Morrison M, White BA, Mackie RI, North American Consortium for Rumen Bacteria, Coutinho PM, Henrissat B, Nelson KE (2010) Comparative genome analysis of Prevotella ruminicola and Prevotella bryantii: insights into their environmental niche. Microb Ecol 60:721–729. https://doi.org/10.1007/s00248-010-9692-8 CrossRefGoogle Scholar
- Schnürer A, Jarvis Å (2018) Microbiology of the biogas process. ISBN 978-91-576-9546-8Google Scholar
- Su XL, Tian Q, Zhang J, Yuan XZ, Shi XS, Guo RB, Qiu YL (2014) Acetobacteroides hydrogenigenes gen. nov., sp. nov., an anaerobic hydrogen-producing bacterium in the family Rikenellaceae isolated from a reed swamp. Int J Syst Evol Microbiol 64:2986–2991. https://doi.org/10.1099/ijs.0.063917-0 CrossRefGoogle Scholar
- Taubert M, Jehmlich N, Vogt C, Richnow HH, Schmidt F, von Bergen M, Seifert J (2011) Time resolved protein-based stable isotope probing (protein-SIP) analysis allows quantification of induced proteins in substrate shift experiments. Proteomics 11:2265–2274. https://doi.org/10.1002/pmic.201000788 CrossRefGoogle Scholar
- Vignais PM, Billoud B, Meyer J (2001) Classification and phylogeny of hydrogenases. FEMS Microbiol Rev 25:455–501. https://doi.org/10.1111/j.1574-6976.2001.tb00587.x CrossRefGoogle Scholar
- Ward B (2015) Chapter 11 – bacterial energy metabolism. In: Tang YW, Sussman M, Liu D, Poxton I (eds) Molecular medical microbiology, 2nd edn. Elsevier, Volume 1, pp 201–233. https://doi.org/10.1016/B978-0-12-397169-2.00011-1
- Yamada T, Sekiguchi Y, Hanada S, Imachi H, Ohashi A, Harada H, Kamagata Y (2006) Anaerolinea thermolimosa sp. nov., Levilinea saccharolytica gen. nov., sp. nov. and Leptolinea tardivitalis gen. nov., sp. nov., novel filamentous anaerobes, and description of the new classes Anaerolineae classis nov. and Caldilineae classis nov. in the bacterial phylum Chloroflexi. Int J Syst Evol Microbiol 56:1331–1340. https://doi.org/10.1099/ijs.0.64169-0 CrossRefGoogle Scholar
- Ziganshin AM, Schmidt T, Lv Z, Liebetrau J, Richnow HH, Kleinsteuber S, Nikolausz M (2016a) Reduction of the hydraulic retention time at constant high organic loading rate to reach the microbial limits of anaerobic digestion in various reactor systems. Bioresour Technol 217:62–71. https://doi.org/10.1016/j.biortech.2016.01.096 CrossRefGoogle Scholar