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Methanobacterium espanolae , an acidiphilic methanogen, required acetate for maximal growth on H2 -CO2 . In the presence of 5 to 15 mM acetate, at a growth pH of 5.5, the μmax was 0.05 h-1 .M. espanolae consumed 12.3 mM acetate during 96 h of incubation at 35°C with shaking at 100 rpm. At initial acetate levels of 2.5 to 10.0 mM, the amount of biomass produced was dependent on the amount of acetate in the medium.13 C nuclear magnetic resonance spectra of protein hydrolysates obtained from cultures grown on [1-13 C]- or [2-13 C]acetate indicated that an incomplete tricarboxylic acid pathway, operating in the reductive direction, was functional in this methanogen. The amino acids were labeled with a very high degree of specificity and at greater than 90% enrichment levels. Less than 2% label randomization occurred between positions primarily labeled from either the carboxyl or methyl group of acetate, and very little label was transferred to positions primarily labeled from CO2 . The labeling pattern of carbohydrates was typical for glucogenesis from pyruvate. This methanogen, by virtue of the properties described above and its ability to incorporate all of the available acetate (10 mM or lower) from the growth medium, has advantages over other microorganisms for use in the production of specifically labeled compounds.

applied and environmental microbiology

Production of Specifically Labeled Compounds by Methanobacterium espanolae Grown on H 2 -CO 2 plus [ 13 C]Acetate