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Ono_etal_AOM_R1_clean.pdf (756.83 kB)

Clumped Isotopologue Fractionation by Microbial Cultures Performing the Anaerobic Oxidation of Methane

revised on 31.08.2020, 12:51 and posted on 01.09.2020, 05:11 by Shuhei Ono, Jeemin H. Rhim, Danielle S. Gruen, Heidi Taubner, Martin Kölling, Gunter Wegener
Methane is abundant in marine subsurface sediments, sourced from microbial or thermocatalytic products. The relative composition of its isotopologues (12CH4, 13CH4, 12CH3D and 13CH3D) is used to infer its sources and sinks. The anaerobic oxidation of methane (AOM) is an important methane sink reaction carried out by consortia of
anaerobic methanotrophic archaea (ANME) and partner bacteria in the presence of methane and sulfate. We investigated the methane isotopologue fractionations during
AOM in experiments with cultures of ANME-1 archaea and partner bacteria obtained from hydrothermally heated gas-rich sediments of the Guaymas Basin. During partial methane consumption in four sets of experiments, residual methane became enriched in 13CH4 and 12CH3D, following kinetic fractionations from 11.1 to 18.3 ‰ and from 117 to 180 ‰,respectively. Results from one set of experiments with D-depleted medium water (δD = – 200‰, whereas the control was –55‰) are inconclusive regarding the reversibility of AOM, which would lead to equilibrium as opposed to kinetic fractionations. The value of Δ13CH3D (the abundance of 13CH3D with respect to that expected from stochastic distribution) increased toward and beyond (up to 8.4‰) the value expected for isotopologue equilibrium (5.3‰ at 37 °C). The kinetic clumped isotopologue fractionation (difference between 13CH3D/12CH3D and 13CH4/12CH4 fractionations) of 4.8 to 12.8 ‰ is in contrast with our previous observation of little to no clumped isotopologue effect during aerobic methane oxidation. Our results demonstrate that AOM can contribute to near
equilibrium Δ13CH3D values observed in marine sediments and 13CH3D systematics can be used to distinguish aerobic versus anaerobic methanotrophic processes in nature.


NASA Astrobiology NNA15BB02A

NSF-Geobiology and Low Temperature Geochemistry, EAR-1852946

DFG cluster of Excellence EXC 2077


Email Address of Submitting Author


Massachusetts Institute of Technology


United States of America

ORCID For Submitting Author


Declaration of Conflict of Interest

The authors declare no competing financial interests.