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Title: Monodeuterated Methane, an Isotopic Tool To Assess Biological Methane Metabolism Rates

Abstract

Biological methane oxidation is a globally relevant process that mediates the flux of an important greenhouse gas through both aerobic and anaerobic metabolic pathways. However, measuring these metabolic rates presents many obstacles, from logistical barriers to regulatory hurdles and poor precision. Here we present a new approach for investigating microbial methane metabolism based on hydrogen atom dynamics, which is complementary to carbon-focused assessments of methanotrophy. The method uses monodeuterated methane (CH 3D) as a metabolic substrate, quantifying the aqueous D/H ratio over time using off-axis integrated cavity output spectroscopy. This approach represents a nontoxic, comparatively rapid, and straightforward approach that supplements existing radiotopic and stable carbon isotopic methods; by probing hydrogen atoms, it offers an additional dimension for examining rates and pathways of methane metabolism. We provide direct comparisons between the CH 3D procedure and the well-established 14CH 4 radiotracer method for several methanotrophic systems, including type I and II aerobic methanotroph cultures and methane-seep sediment slurries and carbonate rocks under anoxic and oxic incubation conditions. In all applications tested, methane consumption values calculated via the CH 3D method were directly and consistently proportional to 14C radiolabel-derived methane oxidation rates. We also employed this method in a nontraditional experimental setup,more » using flexible, gas-impermeable bags to investigate the role of pressure on seep sediment methane oxidation rates. Results revealed an 80% increase over atmospheric pressure in methanotrophic rates the equivalent of ~900-m water depth, highlighting the importance of this parameter on methane metabolism and exhibiting the flexibility of the newly described method.« less

Authors:
 [1];  [1];  [2];  [1];  [1];  [3];  [1];  [4]
  1. California Inst. of Technology (CalTech), Pasadena, CA (United States). Division of Geological and Planetary Sciences
  2. Univ. of Southern California, Los Angeles, CA (United States). Dept. of Biological Science
  3. Harvard Univ., Cambridge, MA (United States), Dept. of Human Evolutionary Biology
  4. USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States); editor
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Oak Ridge Leadership Computing Facility (OLCF)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); National Aeronautics and Space Administration (NASA); Gordon and Betty Moore Foundation (GBMF)
OSTI Identifier:
1565620
Grant/Contract Number:  
SC001057
Resource Type:
Accepted Manuscript
Journal Name:
mSphere
Additional Journal Information:
Journal Volume: 2; Journal Issue: 4; Journal ID: ISSN 2379-5042
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Microbiology

Citation Formats

Marlow, Jeffrey J., Steele, Joshua A., Ziebis, Wiebke, Scheller, Silvan, Case, David, Reynard, Linda M., Orphan, Victoria J., and Green Tringe, Susannah. Monodeuterated Methane, an Isotopic Tool To Assess Biological Methane Metabolism Rates. United States: N. p., 2017. Web. doi:10.1128/mspheredirect.00309-17.
Marlow, Jeffrey J., Steele, Joshua A., Ziebis, Wiebke, Scheller, Silvan, Case, David, Reynard, Linda M., Orphan, Victoria J., & Green Tringe, Susannah. Monodeuterated Methane, an Isotopic Tool To Assess Biological Methane Metabolism Rates. United States. doi:10.1128/mspheredirect.00309-17.
Marlow, Jeffrey J., Steele, Joshua A., Ziebis, Wiebke, Scheller, Silvan, Case, David, Reynard, Linda M., Orphan, Victoria J., and Green Tringe, Susannah. Wed . "Monodeuterated Methane, an Isotopic Tool To Assess Biological Methane Metabolism Rates". United States. doi:10.1128/mspheredirect.00309-17. https://www.osti.gov/servlets/purl/1565620.
@article{osti_1565620,
title = {Monodeuterated Methane, an Isotopic Tool To Assess Biological Methane Metabolism Rates},
author = {Marlow, Jeffrey J. and Steele, Joshua A. and Ziebis, Wiebke and Scheller, Silvan and Case, David and Reynard, Linda M. and Orphan, Victoria J. and Green Tringe, Susannah},
abstractNote = {Biological methane oxidation is a globally relevant process that mediates the flux of an important greenhouse gas through both aerobic and anaerobic metabolic pathways. However, measuring these metabolic rates presents many obstacles, from logistical barriers to regulatory hurdles and poor precision. Here we present a new approach for investigating microbial methane metabolism based on hydrogen atom dynamics, which is complementary to carbon-focused assessments of methanotrophy. The method uses monodeuterated methane (CH3D) as a metabolic substrate, quantifying the aqueous D/H ratio over time using off-axis integrated cavity output spectroscopy. This approach represents a nontoxic, comparatively rapid, and straightforward approach that supplements existing radiotopic and stable carbon isotopic methods; by probing hydrogen atoms, it offers an additional dimension for examining rates and pathways of methane metabolism. We provide direct comparisons between the CH3D procedure and the well-established14CH4 radiotracer method for several methanotrophic systems, including type I and II aerobic methanotroph cultures and methane-seep sediment slurries and carbonate rocks under anoxic and oxic incubation conditions. In all applications tested, methane consumption values calculated via the CH3D method were directly and consistently proportional to14C radiolabel-derived methane oxidation rates. We also employed this method in a nontraditional experimental setup, using flexible, gas-impermeable bags to investigate the role of pressure on seep sediment methane oxidation rates. Results revealed an 80% increase over atmospheric pressure in methanotrophic rates the equivalent of ~900-m water depth, highlighting the importance of this parameter on methane metabolism and exhibiting the flexibility of the newly described method.},
doi = {10.1128/mspheredirect.00309-17},
journal = {mSphere},
number = 4,
volume = 2,
place = {United States},
year = {2017},
month = {8}
}

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