Metagenomic analysis of phosphorus removing sludge communities

Enhanced Biological Phosphorus Removal (EBPR) is not well understood at the metabolic level despite being one of the best-studied microbially-mediated industrial processes due to its ecological and economic relevance. Here we present a metagenomic analysis of two lab-scale EBPR sludges dominated by the uncultured bacterium, Candidatus Accumulibacter phosphatis. This analysis resolves several controversies in EBPR metabolic models and provides hypotheses explaining the dominance of A. phosphatis in this habitat, its lifestyle outside EBPR and probable cultivation requirements. Comparison of the same species from different EBPR sludges highlights recent evolutionary dynamics in the A. phosphatis genome that could be linked to mechanisms for environmental adaptation. In spite of an apparent lack of phylogenetic overlap in the flanking communities of the two sludges studied, common functional themes were found, at least one of them complementary to the inferred metabolism of the dominant organism. The present study provides a much-needed blueprint for a systems-level understanding of EBPR and illustrates that metagenomics enables detailed, often novel, insights into even well-studied biological systems.

[1]  A. Zehnder,et al.  Anaerobic metabolism of bacteria performing enhanced biological phosphate removal , 2000 .

[2]  K. Schleifer,et al.  ARB: a software environment for sequence data. , 2004, Nucleic acids research.

[3]  A. Harden Bacterial Metabolism , 1930, Nature.

[4]  M. Lidstrom,et al.  Identification of Genes Involved in the Glyoxylate Regeneration Cycle in Methylobacterium extorquens AM1, Including Two New Genes, meaC and meaD , 2005, Journal of bacteriology.

[5]  A. Kornberg,et al.  Inorganic polyphosphate: a molecule of many functions. , 1999, Progress in molecular and subcellular biology.

[6]  Tommi S. Jaakkola,et al.  Fast optimal leaf ordering for hierarchical clustering , 2001, ISMB.

[7]  K. Hossner,et al.  Cellular and molecular biology. , 2005 .

[8]  W. McCleary,et al.  The Unphosphorylated Receiver Domain of PhoB Silences the Activity of Its Output Domain , 2000, Journal of bacteriology.

[9]  Paramvir S. Dehal,et al.  Whole-Genome Shotgun Assembly and Analysis of the Genome of Fugu rubripes , 2002, Science.

[10]  B. Burgess,et al.  Evidence for the Direct Interaction of the nifW Gene Product with the MoFe Protein , 1996, The Journal of Biological Chemistry.

[11]  Juan Aguilar,et al.  The Gene yjcG, Cotranscribed with the Gene acs, Encodes an Acetate Permease in Escherichia coli , 2003, Journal of bacteriology.

[12]  Yilin Hu,et al.  Characterization of Azotobacter vinelandii nifZ Deletion Strains , 2004, Journal of Biological Chemistry.

[13]  J. Keasling,et al.  Polyphosphate kinase genes from activated sludge carrying out enhanced biological phosphorus removal. , 2002, Water science and technology : a journal of the International Association on Water Pollution Research.

[14]  A. Kornberg,et al.  Novel Assay Reveals Multiple Pathways Regulating Stress-Induced Accumulations of Inorganic Polyphosphate in Escherichia coli , 1998, Journal of bacteriology.

[15]  Motoharu Onuki,et al.  The microbiology of biological phosphorus removal in activated sludge systems. , 2003, FEMS microbiology reviews.

[16]  Chris Sander,et al.  CAST: an iterative algorithm for the complexity analysis of sequence tracts , 2000, Bioinform..

[17]  J. Peccia,et al.  Involvement of Rhodocyclus-Related Organisms in Phosphorus Removal in Full-Scale Wastewater Treatment Plants , 2002, Applied and Environmental Microbiology.

[18]  A. Kornberg,et al.  Guanosine Tetra- and Pentaphosphate Promote Accumulation of Inorganic Polyphosphate in Escherichia coli * , 1997, The Journal of Biological Chemistry.

[19]  A. Steinbüchel,et al.  Biochemical and genetic analysis of PHA synthases and other proteins required for PHA synthesis. , 1999, International journal of biological macromolecules.

[20]  W. Gujer,et al.  Intracellular carbon flow in phosphorus accumulating organisms from activated sludge systems , 1997 .

[21]  J. Banfield,et al.  Community structure and metabolism through reconstruction of microbial genomes from the environment , 2004, Nature.

[22]  G T Daigger,et al.  Stoichiometry and kinetics of acetate uptake under anaerobic conditions by an enriched culture of phosphorus-accumulating organisms at different pHs. , 2001, Biotechnology and bioengineering.

[23]  S. Tringe,et al.  Comparative Metagenomics of Microbial Communities , 2004, Science.

[24]  F. Tabita,et al.  Aerobic chemolithoautotrophic growth and RubisCO function in Rhodobacter capsulatus and a spontaneous gain of function mutant of Rhodobacter sphaeroides , 1998, Archives of Microbiology.

[25]  B. Wanner,et al.  Involvement of the Escherichia coli phn (psiD) gene cluster in assimilation of phosphorus in the form of phosphonates, phosphite, Pi esters, and Pi , 1991, Journal of bacteriology.

[26]  Thomas L. Madden,et al.  Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. , 1997, Nucleic acids research.

[27]  H. Piaggio Mathematical Analysis , 1955, Nature.

[28]  A Kornberg,et al.  Role of Inorganic Polyphosphate in Promoting Ribosomal Protein Degradation by the Lon Protease in E. coli , 2001, Science.

[29]  D. Jenkins,et al.  Enhanced Biological Phosphorus Removal from Wastewater by Biomass with Different Phosphorus Contents, Part I: Experimental Results and Comparison with Metabolic Models , 2003, Water environment research : a research publication of the Water Environment Federation.

[30]  P. Cullen,et al.  Sequence, genetic, and lacZ fusion analyses of a nifR3–ntrB–ntrC operon in Rhodobacter capsulatus , 1993, Molecular microbiology.

[31]  Aaron Marc Saunders,et al.  Competition between polyphosphate and glycogen accumulating organisms in enhanced biological phosphorus removal systems with acetate and propionate as carbon sources. , 2006, Journal of biotechnology.

[32]  J. R. van der Meer,et al.  Enrichment, phylogenetic analysis and detection of a bacterium that performs enhanced biological phosphate removal in activated sludge. , 1999, Systematic and applied microbiology.

[33]  C. Ouzounis,et al.  Genome-wide identification of genes likely to be involved in human genetic disease. , 2004, Nucleic acids research.

[34]  Alex Bateman,et al.  QuickTree: building huge Neighbour-Joining trees of protein sequences , 2002, Bioinform..

[35]  O. Lenz,et al.  The H2 Sensor of Ralstonia eutropha Is a Member of the Subclass of Regulatory [NiFe] Hydrogenases , 2000, Journal of bacteriology.

[36]  D. White The Physiology and Biochemistry of Prokaryotes , 1999 .

[37]  H. Santos,et al.  Model for carbon metabolism in biological phosphorus removal processes based on in vivo13C-NMR labelling experiments , 1996 .

[38]  Inna Dubchak,et al.  The integrated microbial genomes (IMG) system , 2005, Nucleic Acids Res..

[39]  T. Z. DeSantis,et al.  Comprehensive aligned sequence construction for automated design of effective probes (CASCADE-P) using 16S rDNA , 2003, Bioinform..

[40]  T. Fukui,et al.  Genetic Analysis of Comamonas acidovoransPolyhydroxyalkanoate Synthase and Factors Affecting the Incorporation of 4-Hydroxybutyrate Monomer , 1998, Applied and Environmental Microbiology.

[41]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .