Heike Rütters

Phospholipid analysis as a tool to study complex microbial communities in marine sediments.

To complement information on microbial communities in marine sediments that can be obtained using microbiological methods, we developed an analytical procedure to trace microbial lipids in environmental samples. We focused on analyzing intact phospholipids as these membrane constituents are known to be biomarkers for viable cells. Analysis of intact phospholipids from a fractionated and preconcentrated sediment extract was achieved using liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS). The combined analysis of phospholipid types and their fatty acid substituents allowed a differentiation between various groups of microorganisms living in the sediment. For comparison three strains of marine sulfate-reducing bacteria (SRB) were analysed for their lipid content.

Monoalkylether phospholipids in the sulfate-reducing bacteria Desulfosarcina variabilis and Desulforhabdus amnigenus

Abstract. In this study, cellular lipid compositions of two mesophilic sulfate-reducing bacteria were analyzed by high performance liquid chromatography-mass spectrometry (HPLC-MS). In Desulfosarcina variabilis and Desulforhabdus amnigenus, alkylether-containing phospholipids were detected which had previously only been found in significant amounts in deeply branching hyperthermophilic bacteria and archaea. Combining information from HPLC-MS analysis and chemical degradation experiments, ether lipids were identified as 1-alkyl-2-acyl-phosphatidyl ethanolamines, glycerols and cholines. In Desulforhabdus amnigenus, n-penta-, n-hexa- and n-heptadecyl ethers were present (in order of decreasing abundance), whereas Desulfosarcina variabilis solely contained n-hexadecyl ether side chains.

Microbial communities in a Wadden Sea sediment core: clues from analyses of intact glyceride lipids, and released fatty acids

In this study microbial communities were investigated in a 55 cm sediment core from a tidal flat close to Neuharlingersiel, NW Germany, using bulk and molecular geochemical parameters. We focussed on the analysis of various types of glyceride lipids. In particular, we concentrated on the analysis of intact phospholipids by HPLC-MS(MS) as biomarkers for “viable” microorganisms. Highest abundances of intact microbial lipids were found in the surface layer of the sediment, where phospholipid types and their fatty acid patterns indicated a strong contribution of eukaryotic algae. Microbial lipids appeared to be recycled or degraded rapidly within the uppermost 10 cm of the sediment because abundances of intact phospholipids and triglycerides rapidly decreased with depth. Also, the sedimentary contents of their degradation products (e.g. di- and monoglycerides, free fatty acids) showed a strong decline in the top layers. In the depth interval of 10-20 cm, phospholipid contents and species indicated a high abundance of bacterial biomass. In the deepest samples, intact phospholipids still were detected suggesting the presence of “viable” bacteria at a sediment depth of 50 cm.

Mycosporine-glutamicol-glucoside: a natural UV-absorbing secondary metabolite of rock-inhabiting microcolonial fungi.

Microcolonial ascomycetes are known to inhabit bare rock surfaces in cold and hot deserts and thus are habitually exposed to high levels of solar radiation. Several of these stress-tolerant fungal isolates, cultivated in the laboratory under daylight illumination, were studied for the presence of effective UV-radiation protection substances. Liquid chromatography/mass spectrometry (LC/MS) and liquid chromatography/tandem mass spectrometry (LC/MS/MS) analyses allowed for efficient separation and structure clarification of two mycosporines. It was demonstrated that both mycosporine-glutamicol-glucoside and mycosporine-glutaminol-glucoside are natural and constitutive secondary metabolites of microcolonial fungi. The function and relation of these substances in the fungal cell are discussed.

Novel bacteriochlorophyll e structures and species-specific variability of pigment composition in green sulfur bacteria

Abstract. The relative composition of bacteriochlorophyll (BChl) homologs in five different strains of brown-colored green sulfur bacteria was investigated by HPLC-MS/MS and NMR analyses. In addition, the effect of incubation light intensities on homolog distribution was studied in one of the strains (strain Dagow III). A total of 23 different BChl e structures were detected and comprise four homologous porphyrin ring systems and eight different esterifying alcohols. Several BChl e structures are novel. These include a C-8 ethyl, C-12 methyl [E, M] BChl eF homolog which was identified by 1H-NMR analyses of the isolated, main farnesyl homologs (BChl eF). In addition, five previously unknown homolog series with dodecanol, pentadecenol, tetradecanol, hexadecenol and phytol as the esterifying alcohols were detected. The composition of BChl e homologs from the five strains of green sulfur bacteria differed with respect to the relative abundance of the homologs (BChl eF : 25.6–67.0% of total BChl e content in stationary cultures). In strain Dagow III, the abundance of BChl eF homologs decreased upon entry into the stationary phase. In all free-living strains, the abundance of BChl eF was increased when the relative carotenoid content was low. The present results provide a detailed picture of pigment composition in chlorosomes and thus will help to elucidate their structure and function. Furthermore, the newly discovered BChl e molecules are valuable biomarkers for the study of the occurrence and metabolism of green sulfur bacteria in past and present ecosystems.

Tateyamaria pelophila sp. nov., a facultatively anaerobic alphaproteobacterium isolated from tidal-flat sediment, and emended descriptions of the genus Tateyamaria and of Tateyamaria omphalii

A Gram-negative motile rod, strain SAM4T, was isolated from the highest positive dilution of a most probable number series inoculated with tidal-flat sediments from the German North Sea coast. The isolate grew at 4-35 degrees C and showed constant growth yields throughout almost the whole temperature range. Growth was observed between pH 6 and 9 and at salinities of 0.3-10.2%. Strain SAM4T required Na+ for growth, contained bacteriochlorophyll a and was catalase- and oxidase-positive. It was nutritionally versatile growing on a variety of carbon compounds including carbohydrates, amino acids and organic acids like lactate or succinate. It grew anaerobically on complex media such as marine broth, indicating fermentation, and by reducing trimethylammonium oxide. The dominant phospholipids were phosphatidylethanolamine and phosphatidylglycerol, whereas only traces of phosphatidylcholine and an unidentified lipid were found. The major fatty acid was n-C18:1omega7c. The DNA G+C content was 56.4 mol%. The isolate was identified as a member of the Roseobacter clade within the class Alphaproteobacteria. However, based on phylogenetic, phenotypic and physiological data, it clearly differs from its closest relative Tateyamaria omphalii. Therefore, a novel species is proposed: Tateyamaria pelophila sp. nov., with strain SAM4T (=DSM 17270T=LMG 23018T) as the type strain. Emended descriptions of the genus Tateyamaria and of Tateyamaria omphalii are also presented.