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Title: 15 N- and 2 H proteomic stable isotope probing links nitrogen flow to archaeal heterotrophic activity

Journal Article · · Environmental Microbiology

Understanding how individual species contribute to nutrient transformations in a microbial community is critical to prediction of overall ecosystem function. We conducted microcosm experiments in which floating acid mine drainage (AMD) microbial biofilms were submerged recapitulating the final stage in a natural biofilm life cycle. Biofilms were amended with either 15NH4 + or deuterium oxide (2H2O) and proteomic stable isotope probing (SIP) was used to track the extent to which different members of the community used these molecules in protein synthesis across anaerobic iron-reducing, aerobic iron-reducing and aerobic iron-oxidizing environments. Sulfobacillus spp. synthesized 15N-enriched protein almost exclusively under iron-reducing conditions whereas the Leptospirillum spp. synthesized 15N-enriched protein in all conditions. There were relatively few 15Nenriched archaeal proteins, and all showed low atom% enrichment, consistent with Archaea synthesizing protein using the predominantly 14N biomass derived from recycled biomolecules. In parallel experiments using 2H2O, extensive archaeal protein synthesis was detected in all conditions. In contrast, the bacterial species showed little protein synthesis using 2H2O. The nearly exclusive ability of Archaea to synthesize proteins using 2H2O may be due to archaeal heterotrophy, whereby Archaea off set deleterious effects of 2H by accessing 1H generated byrespiration of organic compounds.

Research Organization:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Oak Ridge Leadership Computing Facility (OLCF)
Sponsoring Organization:
USDOE Office of Science (SC)
DOE Contract Number:
AC05-00OR22725
OSTI ID:
1265333
Journal Information:
Environmental Microbiology, Vol. 16, Issue 10; ISSN 1462-2912
Publisher:
Wiley
Country of Publication:
United States
Language:
English

References (44)

Incorporation of carbon and nitrogen atoms into proteins measured by protein-based stable isotope probing (Protein-SIP) journal September 2008
Growth and Sugar Metabolism of a Thermoaddophilic Iron-oxidizing Mixotrophic Bacterium journal June 1984
Strain-resolved community proteomics reveals recombining genomes of acidophilic bacteria journal March 2007
Cultivation and quantitative proteomic analyses of acidophilic microbial communities journal December 2009
AMD biofilms: using model communities to study microbial evolution and ecological complexity in nature journal February 2010
An appraisal of methods for linking environmental processes to specific microbial taxa journal May 2010
Comparative genomics in acid mine drainage biofilm communities reveals metabolic and structural differentiation of co-occurring archaea journal January 2013
Ferrozine---a new spectrophotometric reagent for iron journal June 1970
Exploring the limits of robust detection of incorporation of 13C by mass spectrometry in protein-based stable isotope probing (protein-SIP) journal August 2011
Quantitative proteomic analyses of the response of acidophilic microbial communities to different pH conditions journal January 2011
Stable-isotope probing as a tool in microbial ecology journal February 2000
eggNOG v2.0: extending the evolutionary genealogy of genes with enhanced non-supervised orthologous groups, species and functional annotations journal November 2009
Insights into the Diversity of Eukaryotes in Acid Mine Drainage Biofilm Communities journal February 2009
Ammonium uptake by heterotrophic bacteria in the Delaware estuary and adjacent coastal waters journal July 1995
Stable isotope probing — linking microbial identity to function journal May 2005
Sipros/ProRata: a versatile informatics system for quantitative community proteomics journal June 2013
Quantitative Tracking of Isotope Flows in Proteomes of Microbial Communities journal February 2011
Community Genomic and Proteomic Analyses of Chemoautotrophic Iron-Oxidizing "Leptospirillum rubarum" (Group II) and "Leptospirillum ferrodiazotrophum" (Group III) Bacteria in Acid Mine Drainage Biofilms journal May 2009
Heterotrophic Archaea Contribute to Carbon Cycling in Low-pH, Suboxic Biofilm Communities journal September 2012
Large-scale analysis of the yeast proteome by multidimensional protein identification technology journal March 2001
Community-wide analysis of microbial genome sequence signatures journal January 2009
Response to oxygen limitation in bacteria of the genus sulfobacillus journal February 2010
Towards determining details of anaerobic growth coupled to ferric iron reduction by the acidophilic archaeon ‘Ferroplasma acidarmanus’ Fer1 journal October 2006
Community structure and metabolism through reconstruction of microbial genomes from the environment journal February 2004
Direct linking of microbial populations to specific biogeochemical processes by 13C-labelling of biomarkers journal April 1998
Posttranslational modification and sequence variation of redox-active proteins correlate with biofilm life cycle in natural microbial communities journal May 2010
In Situ Evolutionary Rate Measurements Show Ecological Success of Recently Emerged Bacterial Hybrids journal April 2012
Protein-based stable isotope probing (Protein-SIP) reveals active species within anoxic mixed cultures journal June 2008
Sulfobacillus benefaciens sp. nov., an acidophilic facultative anaerobic Firmicute isolated from mineral bioleaching operations journal August 2008
Protein-SIP enables time-resolved analysis of the carbon flux in a sulfate-reducing, benzene-degrading microbial consortium journal July 2012
Protein-based stable isotope probing journal November 2010
Limitations in detection of 15N incorporation by mass spectrometry in protein-based stable isotope probing (protein-SIP) journal March 2013
DNA-based stable isotope probing: a link between community structure and function journal June 2009
Studies on incorporation of Deuterium into bacteria journal April 1963
Deuterium isotope effects on noncovalent interactions between molecules journal February 1999
Steady-state analysis of kinetic isotope effects in enzymic reactions journal June 1975
Acid mine drainage biogeochemistry at Iron Mountain, California journal June 2004
Direct Cellular Lysis/Protein Extraction Protocol for Soil Metaproteomics journal December 2010
Decimal Place Slope, A Fast and Precise Method for Quantifying 13 C Incorporation Levels for Detecting the Metabolic Activity of Microbial Species journal January 2010
Characterization of Cytochrome 579, an Unusual Cytochrome Isolated from an Iron-Oxidizing Microbial Community journal May 2008
Evaluation of Multidimensional Chromatography Coupled with Tandem Mass Spectrometry (LC/LC−MS/MS) for Large-Scale Protein Analysis:  The Yeast Proteome journal February 2003
Calculation of partial isotope incorporation into peptides measured by mass spectrometry journal June 2010
New Group in the Leptospirillum Clade: Cultivation-Independent Community Genomics, Proteomics, and Transcriptomics of the New Species “Leptospirillum Group IV UBA BS” journal May 2013
Oxygen-dependent niche formation of a pyrite-dependent acidophilic consortium built by archaea and bacteria journal April 2013