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Applied and Environmental Microbiology, August 2005, p. 4345-4351, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4345-4351.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

In Vitro Study of Lipid Biosynthesis in an Anaerobically Methane-Oxidizing Microbial Mat

Martin Blumenberg,¹ Richard Seifert,^1* Katja Nauhaus,^2, Thomas Pape,¹ and Walter Michaelis¹

Institute of Biogeochemistry and Marine Chemistry, University of Hamburg, Bundesstr. 55, 20146 Hamburg, Germany,¹ Max Planck Institute for Marine Microbiology, Celsiusstr. 1, 28359 Bremen, Germany²

Received 18 November 2004/ Accepted 20 February 2005

The anaerobic oxidation of methane (AOM) is a key process in the global methane cycle, and the majority of methane formed in marine sediments is oxidized in this way. Here we present results of an in vitro ¹³CH₄ labeling study (¹³CH₄, 5,400) in which microorganisms that perform AOM in a microbial mat from the Black Sea were used. During 316 days of incubation, the ¹³C uptake into the mat biomass increased steadily, and there were remarkable differences for individual bacterial and archaeal lipid compounds. The greatest shifts were observed for bacterial fatty acids (e.g., hexadec-11-enoic acid [16:111]; difference between the ¹³C at the start and the end of the experiment [¹³C_start-end], 160). In contrast, bacterial glycerol diethers exhibited only slight changes in ¹³C (¹³C_start-end, 10). Differences were also found for individual archaeal lipids. Relatively high uptake of methane-derived carbon was observed for archaeol (¹³C_start-end, 25), a monounsaturated archaeol, and biphytanes, whereas for sn-2-hydroxyarchaeol there was considerably less change in the ¹³C (¹³C_start-end, 2). Moreover, an increase in the uptake of ¹³C for compounds with a higher number of double bonds within a suite of polyunsaturated 2,6,10,15,19-pentamethyleicosenes indicated that in methanotrophic archaea there is a biosynthetic pathway similar to that proposed for methanogenic archaea. The presence of group-specific biomarkers (for ANME-1 and ANME-2 associations) and the observation that there were differences in ¹³C uptake into specific lipid compounds confirmed that multiple phylogenetically distinct microorganisms participate to various extents in biomass formation linked to AOM. However, the greater ¹³C uptake into the lipids of the sulfate-reducing bacteria (SRB) than into the lipids of archaea supports the hypothesis that there is autotrophic growth of SRB on small methane-derived carbon compounds supplied by the methane oxidizers.

Publication GEOTECH-119 of the GEOTECHNOLOGIEN program of the BMBF and the DFG and publication 14 of the GHOSTDABS research program.

Present address: Department Biology I, Microbiology and Genetics, Ludwig Maximilians University of Munich, Maria-Ward-Str. 1a, 80638 Munich, Germany.

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