Isotopic evidence for biological nitrogen fixation by molybdenum-nitrogenase from 3.2 Gyr

Journal name:
Nature
Volume:
520,
Pages:
666–669
Date published:
DOI:
doi:10.1038/nature14180
Received
Accepted
Published online

Nitrogen is an essential nutrient for all organisms that must have been available since the origin of life. Abiotic processes including hydrothermal reduction1, photochemical reactions2, or lightning discharge3 could have converted atmospheric N2 into assimilable NH4+, HCN, or NOx species, collectively termed fixed nitrogen. But these sources may have been small on the early Earth, severely limiting the size of the primordial biosphere4. The evolution of the nitrogen-fixing enzyme nitrogenase, which reduces atmospheric N2 to organic NH4+, thus represented a major breakthrough in the radiation of life, but its timing is uncertain5, 6. Here we present nitrogen isotope ratios with a mean of 0.0 ± 1.2‰ from marine and fluvial sedimentary rocks of prehnite–pumpellyite to greenschist metamorphic grade between 3.2 and 2.75 billion years ago. These data cannot readily be explained by abiotic processes and therefore suggest biological nitrogen fixation, most probably using molybdenum-based nitrogenase as opposed to other variants that impart significant negative fractionations7. Our data place a minimum age constraint of 3.2 billion years on the origin of biological nitrogen fixation and suggest that molybdenum was bioavailable in the mid-Archaean ocean long before the Great Oxidation Event.

At a glance

Figures

  1. Sedimentary [dgr]15N through time.
    Figure 1: Sedimentary δ15N through time.

    All points labelled ‘Soanesville, ‘Witwatersrand’, ‘Nullagine’, and ‘Fortescue’ are from this study, all others are compiled from the literature. ‘Hydrothermal’ refers to samples from hydrothermal veins; ‘Metamorphosed’, metamorphosed above greenschist facies; ‘Witwatersrand’, Witwatersrand Supergroup. Insert shows data from this study at higher resolution; note that Soanesville Group data at 3.2 Gyr represent 20 superimposed points. See Supplementary Information for references.

  2. Whole-rock [dgr]15N versus TOC.
    Figure 2: Whole-rock δ15N versus TOC.

    Symbols are as in Fig. 1. Error bars are 1σ of two or three replicate analyses of the same decarbonated sample aliquot. Dark blue shading, range of δ15N values expressed in cultures that use the Mo-nitrogenase Nif; light blue, extension of this range seen in some wild types and thermophilic organisms using Nif; green, range observed in cultures using pure Vnf or Anf7, 17, 18. Almost all data points plot within the range of Mo-based nitrogen fixation. Outliers in fluvial sediments may reflect local redox chemistry.

  3. TOC and [dgr]15N versus Mo and V enrichment factors.
    Figure 3: TOC and δ15N versus Mo and V enrichment factors.

    a, TOC; b, δ15N. Enrichment factors are calculated relative to average upper continental crust (Methods). All values are relatively close to 1.0, indicating insignificant authigenic enrichments of V or Mo in sediments. No trends are observed, indicating that availability of Mo and V did not influence either TOC or δ15N.

  4. Organic carbon (a) and bulk nitrogen (b) isotopes as a function of total abundances.
    Extended Data Fig. 1: Organic carbon (a) and bulk nitrogen (b) isotopes as a function of total abundances.

    Correlation coefficients refer to data points and trend lines of the same colour. See Supplementary Information 3 for discussion. Each sample was analysed in two or three replicates.

  5. Crust-normalized elemental abundances.
    Extended Data Fig. 2: Crust-normalized elemental abundances.

    a, Normalized relative to average upper continental crust; b, normalized relative to average upper Archaean crust (Methods). Some data points in b are missing because concentrations for those elements in average upper Archaean crust were not provided. In both parts, elements are sorted by relative abundance in upper continental crust.

  6. Reproducibility of the UW-McRae in-house standard over a range of sample sizes.
    Extended Data Fig. 3: Reproducibility of the UW-McRae in-house standard over a range of sample sizes.

    Filled diamonds, analysed with regular CN method; grey circles, analysed with a soda-lime column to scrub CO2 from the gas stream; horizontal lines, total average (solid) ± 1 s.d. (dashed).

Tables

  1. Comparison of kerogen and whole-rock data for samples with sufficient material from the Soanesville Group and the Witwatersrand Supergroup
    Extended Data Table 1: Comparison of kerogen and whole-rock data for samples with sufficient material from the Soanesville Group and the Witwatersrand Supergroup

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Author information

  1. Present address: Mineral Services, SGS South Africa, 58 Melville Street, Booysens, Johannesburg 2091, South Africa.

    • Bradley M. Guy

Affiliations

  1. Department of Earth & Space Sciences and Astrobiology Program, University of Washington, Seattle, Washington 98195-1310, USA

    • Eva E. Stüeken,
    • Roger Buick &
    • Matthew C. Koehler
  2. Department of Geology, University of Johannesburg, Auckland Park 2006, South Africa

    • Bradley M. Guy

Contributions

R.B. collected samples from the Soanesville Group; B.M.G. collected samples from the Witwatersrand Supergroup; R.B. and E.E.S. collected samples from the Fortescue Group and Nullagine Group; E.E.S. and M.C.K. performed the analyses; E.E.S. drafted the manuscript with advice and contributions from R.B.; B.M.G. and M.C.K. commented on the draft.

Competing financial interests

The authors declare no competing financial interests.

Corresponding author

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Author details

Extended data figures and tables

Extended Data Figures

  1. Extended Data Figure 1: Organic carbon (a) and bulk nitrogen (b) isotopes as a function of total abundances. (145 KB)

    Correlation coefficients refer to data points and trend lines of the same colour. See Supplementary Information 3 for discussion. Each sample was analysed in two or three replicates.

  2. Extended Data Figure 2: Crust-normalized elemental abundances. (232 KB)

    a, Normalized relative to average upper continental crust; b, normalized relative to average upper Archaean crust (Methods). Some data points in b are missing because concentrations for those elements in average upper Archaean crust were not provided. In both parts, elements are sorted by relative abundance in upper continental crust.

  3. Extended Data Figure 3: Reproducibility of the UW-McRae in-house standard over a range of sample sizes. (104 KB)

    Filled diamonds, analysed with regular CN method; grey circles, analysed with a soda-lime column to scrub CO2 from the gas stream; horizontal lines, total average (solid) ± 1 s.d. (dashed).

Extended Data Tables

  1. Extended Data Table 1: Comparison of kerogen and whole-rock data for samples with sufficient material from the Soanesville Group and the Witwatersrand Supergroup (181 KB)

Supplementary information

PDF files

  1. Supplementary Information (832 KB)

    This file contains Supplementary Information 1-3, Supplementary Tables 1-4 and additional references.

Additional data