Establishment and metabolic analysis of a model microbial community for understanding trophic and electron accepting interactions of subsurface anaerobic environments

BackgroundCommunities of microorganisms control the rates of key biogeochemical cycles, and are important for biotechnology, bioremediation, and industrial microbiological processes. For this reason, we constructed a model microbial community comprised of three species dependent on trophic interactions. The three species microbial community was comprised of Clostridium cellulolyticum, Desulfovibrio vulgaris Hildenborough, and Geobacter sulfurreducens and was grown under continuous culture conditions. Cellobiose served as the carbon and energy source for C. cellulolyticum, whereas D. vulgaris and G. sulfurreducens derived carbon and energy from the metabolic products of cellobiose fermentation and were provided with sulfate and fumarate respectively as electron acceptors.ResultsqPCR monitoring of the culture revealed C. cellulolyticum to be dominant as expected and confirmed the presence of D. vulgaris and G. sulfurreducens. Proposed metabolic modeling of carbon and electron flow of the three-species community indicated that the growth of C. cellulolyticum and D. vulgaris were electron donor limited whereas G. sulfurreducens was electron acceptor limited.ConclusionsThe results demonstrate that C. cellulolyticum, D. vulgaris, and G. sulfurreducens can be grown in coculture in a continuous culture system in which D. vulgaris and G. sulfurreducens are dependent upon the metabolic byproducts of C. cellulolyticum for nutrients. This represents a step towards developing a tractable model ecosystem comprised of members representing the functional groups of a trophic network.

[1]  F. Glöckner,et al.  Succinate dehydrogenase functioning by a reverse redox loop mechanism and fumarate reductase in sulphate-reducing bacteria. , 2006, Microbiology.

[2]  G. Falony,et al.  Coculture Fermentations of Bifidobacterium Species and Bacteroides thetaiotaomicron Reveal a Mechanistic Insight into the Prebiotic Effect of Inulin-Type Fructans , 2009, Applied and Environmental Microbiology.

[3]  Michael Wagner,et al.  Wastewater treatment: a model system for microbial ecology. , 2006, Trends in biotechnology.

[4]  G. Hardin The competitive exclusion principle. , 1960, Science.

[5]  Daniel B. Oerther,et al.  A vista for microbial ecology and environmental biotechnology. , 2006, Environmental science & technology.

[6]  Sze-Bi Hsu,et al.  A Mathematical Theory for Single-Nutrient Competition in Continuous Cultures of Micro-Organisms , 1977 .

[7]  R. Y. Morita,et al.  Bioavailability of energy and its relationship to growth and starvation survival in nature , 1988 .

[8]  Feng Gao,et al.  Microbial biodegradation of polyaromatic hydrocarbons. , 2008, FEMS microbiology reviews.

[9]  D. Stahl,et al.  Molecular Systems Biology 3; Article number 92; doi:10.1038/msb4100131 Citation: Molecular Systems Biology 3:92 , 2022 .

[10]  Abraham Esteve-Núñez,et al.  Growth of Geobacter sulfurreducens under nutrient-limiting conditions in continuous culture. , 2005, Environmental microbiology.

[11]  Kenneth H. Williams,et al.  Proteogenomic Monitoring of Geobacter Physiology during Stimulated Uranium Bioremediation , 2009, Applied and Environmental Microbiology.

[12]  Rekha Seshadri,et al.  The genome sequence of the anaerobic, sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough , 2004, Nature Biotechnology.

[13]  D. Lovley,et al.  Quantification of Desulfovibrio vulgaris Dissimilatory Sulfite Reductase Gene Expression during Electron Donor- and Electron Acceptor-Limited Growth , 2008, Applied and Environmental Microbiology.

[14]  E. LeBoeuf,et al.  Fluorescence spectroscopic studies of natural organic matter fractions. , 2003, Chemosphere.

[15]  Bernhard Schink,et al.  Synergistic interactions in the microbial world , 2002, Antonie van Leeuwenhoek.

[16]  A. K. Haritash,et al.  Biodegradation aspects of polycyclic aromatic hydrocarbons (PAHs): a review. , 2009, Journal of hazardous materials.

[17]  G. Tullock,et al.  Competitive Exclusion. , 1960, Science.

[18]  A. Arkin,et al.  The Electron Transfer System of Syntrophically Grown Desulfovibrio vulgaris , 2009, Journal of bacteriology.

[19]  D. Bagley,et al.  Supersaturation of dissolved H(2) and CO (2) during fermentative hydrogen production with N(2) sparging. , 2006, Biotechnology letters.

[20]  G. Macfarlane,et al.  Models for intestinal fermentation: association between food components, delivery systems, bioavailability and functional interactions in the gut. , 2007, Current opinion in biotechnology.

[21]  H. Harms,et al.  Mass transfer limitation of microbial growth and pollutant degradation , 1997, Journal of Industrial Microbiology and Biotechnology.

[22]  Suiying Huang,et al.  How Stable Is Stable? Function versus Community Composition , 1999, Applied and Environmental Microbiology.

[23]  Robin F. Harris,et al.  Determination of the Carbon-Bound Electron Composition of Microbial Cells and Metabolites by Dichromate Oxidation , 1979, Applied and environmental microbiology.

[24]  Derek R. Lovley,et al.  Genetic Characterization of a Single Bifunctional Enzyme for Fumarate Reduction and Succinate Oxidation in Geobacter sulfurreducens and Engineering of Fumarate Reduction in Geobacter metallireducens , 2006, Journal of bacteriology.

[25]  H. Kent,et al.  The genomes of Desulfovibrio gigas and D. vulgaris. , 1984, Journal of general microbiology.

[26]  D. Lovley,et al.  Geobacter sulfurreducens sp. nov., a hydrogen- and acetate-oxidizing dissimilatory metal-reducing microorganism , 1994, Applied and environmental microbiology.

[27]  R. Amann,et al.  Dual staining of natural bacterioplankton with 4',6-diamidino-2-phenylindole and fluorescent oligonucleotide probes targeting kingdom-level 16S rRNA sequences , 1992, Applied and environmental microbiology.

[28]  Judy D. Wall,et al.  Analysing the Metabolic Capabilities of Desulfovibrio Species through Genetic Manipulation , 2006, Biotechnology & genetic engineering reviews.

[29]  Marc Viñas,et al.  Culture-dependent and -independent approaches establish the complexity of a PAH-degrading microbial consortium. , 2005, Canadian journal of microbiology.

[30]  Thomas Egli,et al.  The Ecological and Physiological Significance of the Growth of Heterotrophic Microorganisms with Mixtures of Substrates , 1995 .

[31]  Jizhong Zhou,et al.  Microbial Communities in Contaminated Sediments, Associated with Bioremediation of Uranium to Submicromolar Levels , 2008, Applied and Environmental Microbiology.

[32]  Y. Li,et al.  Long-term performance of bioreactors cleaning mercury-contaminated wastewater and their response to temperature and mercury stress and mechanical perturbation. , 2001, Biotechnology and bioengineering.

[33]  S. Macnaughton,et al.  Diversity and Characterization of Sulfate-Reducing Bacteria in Groundwater at a Uranium Mill Tailings Site , 2001, Applied and Environmental Microbiology.

[34]  M. Desvaux Unravelling Carbon Metabolism in Anaerobic Cellulolytic Bacteria , 2006, Biotechnology progress.

[35]  P. Long,et al.  Characterization of Microbial Activities and U Reduction in a Shallow Aquifer Contaminated by Uranium Mill Tailings , 2003, Microbial Ecology.

[36]  Byoung-Chan Kim,et al.  Insights into genes involved in electricity generation in Geobacter sulfurreducens via whole genome microarray analysis of the OmcF-deficient mutant. , 2008, Bioelectrochemistry.

[37]  T. D. Brock,et al.  Microbial Life at 90 C: the Sulfur Bacteria of Boulder Spring , 1971, Journal of bacteriology.

[38]  Robert W. Hoioarth A rapid and precise method for determining sulfate seawater, estuarine waters, and sediment pore waters1 , 1978 .

[39]  M. Desvaux,et al.  Carbon Flux Distribution and Kinetics of Cellulose Fermentation in Steady-State Continuous Cultures of Clostridium cellulolyticum on a Chemically Defined Medium , 2001, Journal of bacteriology.

[40]  D. Lovley,et al.  Preferential Reduction of Fe(III) over Fumarate by Geobacter sulfurreducens , 2004, Journal of bacteriology.

[41]  Grigoriy E. Pinchuk,et al.  The influence of cultivation methods on Shewanella oneidensis physiology and proteome expression , 2007, Archives of Microbiology.

[42]  D. Bagley,et al.  Supersaturation of Dissolved H2 and CO2 During Fermentative Hydrogen Production with N2 Sparging , 2006, Biotechnology Letters.

[43]  T. Egli,et al.  Growth Kinetics of Suspended Microbial Cells: From Single-Substrate-Controlled Growth to Mixed-Substrate Kinetics , 1998, Microbiology and Molecular Biology Reviews.

[44]  J. Wall,et al.  Development of a Markerless Genetic Exchange System for Desulfovibrio vulgaris Hildenborough and Its Use in Generating a Strain with Increased Transformation Efficiency , 2009, Applied and Environmental Microbiology.

[45]  T. Phelps,et al.  Comparison between geochemical and biological estimates of subsurface microbial activities , 2004, Microbial Ecology.

[46]  J A Eisen,et al.  Genome of Geobacter sulfurreducens: Metal Reduction in Subsurface Environments , 2003, Science.

[47]  L. Raskin,et al.  Diversity and dynamics of microbial communities in engineered environments and their implications for process stability. , 2003, Current opinion in biotechnology.

[48]  D. Balkwill,et al.  Change in Bacterial Community Structure during In Situ Biostimulation of Subsurface Sediment Cocontaminated with Uranium and Nitrate , 2004, Applied and Environmental Microbiology.

[49]  E. LeBoeuf,et al.  Spectroscopic characterization of the structural and functional properties of natural organic matter fractions. , 2002, Chemosphere.

[50]  R. Lenski,et al.  Coexistence of two competitors on one resource and one inhibitor: a chemostat model based on bacteria and antibiotics. , 1986, Journal of theoretical biology.

[51]  R. Knight,et al.  The Human Microbiome Project , 2007, Nature.

[52]  M. Desvaux,et al.  Carbon and Electron Flow in Clostridium cellulolyticum Grown in Chemostat Culture on Synthetic Medium , 1999, Journal of bacteriology.

[53]  E. N. Bjordal,et al.  Major hazards in the process industries: Achievements and challenges in loss prevention* , 1992 .

[54]  Michael Wagner,et al.  Bacterial community composition and function in sewage treatment systems. , 2002, Current opinion in biotechnology.

[55]  Donald R. Metzler,et al.  Stimulating the In Situ Activity of Geobacter Species To Remove Uranium from the Groundwater of a Uranium-Contaminated Aquifer , 2003, Applied and Environmental Microbiology.