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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{dagger}

Martin Blumenberg,1 Richard Seifert,1* Katja Nauhaus,2,{ddagger} 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 ({delta}13CH4, ~5,400{per thousand}) 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 [16:1{Delta}11]; difference between the {delta}13C at the start and the end of the experiment [{Delta}{delta}13Cstart-end], ~160{per thousand}). In contrast, bacterial glycerol diethers exhibited only slight changes in {delta}13C ({Delta}{delta}13Cstart-end, ~10{per thousand}). Differences were also found for individual archaeal lipids. Relatively high uptake of methane-derived carbon was observed for archaeol ({Delta}{delta}13Cstart-end, ~25{per thousand}), a monounsaturated archaeol, and biphytanes, whereas for sn-2-hydroxyarchaeol there was considerably less change in the {delta}13C ({Delta}{delta}13Cstart-end, ~2{per thousand}). 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.

* Corresponding author. Mailing address: Institute of Biogeochemistry and Marine Chemistry, University of Hamburg, Bundesstr. 55, 20146 Hamburg, Germany. Phone: 494 04283 84987. Fax: 494 04283 86347. E-mail: seifert{at}geowiss.uni-hamburg.de.

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

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

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.



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