Rudolf Hartmann

Structural determination of N-2′-hydroxyoctadecenoyl-1-O-β-d-glucopyranosyl-9-methyl-4, 8-sphingadienine from species of Aspergillus

Ceramide monohexosides from Aspergillus fumigatus 2140 and 2109 strains and Aspergillus versicolor 550 strain, obtained by silica gel 60, and Iatrobeads chromatography were analysed using high-resolution 1D-, 2D-1H-NMR and 13C-NMR spectroscopy and fast atom bombardment mass spectrometry (FAB-MS). The ceramide monohexoside fraction (CMH) from A. fumigatus 2140 and A. versicolor 550 was identified as glucosylceramide, whereas glucose and galactose were present at a ratio of 1:1 in the CMH of A. fumigatus 2109. The major glycosphingolipid has a particular ceramide composition consisting of 9-methyl-4,8-sphingadienine linked to a 2-hydroxyoctadec-3-enoic acid. Although the structures presently described are similar to those of monohexosylceramides from other fungi, including edible ones, this is the first report on their occurrence in species pathogenic in humans.

The aqueous solution structure of a lipoteichoic acid from Streptococcus pneumoniae strain R6 containing 2,4-diamino-2,4,6-trideoxy-galactose: evidence for conformational mobility of the galactopyranose ring

The 2D-NOESY spectra for the per-N-acetylated and the native lipoteichoic acid (LTA) oligomer from Streptococcus pneumoniae strain R6 clearly indicate a difference in conformation of the 2,4,6-trideoxy-galactopyranose ring. Whereas the 2,4-N-acetylated Gal24N adopts the usual 4C1 chair conformation, the native 2-N-acetyl-4-amino Gal24N exhibits conformational mobility with comparable populations in the 4C1 chair and 5S1 skew conformations, as determined using MD simulation for the partial trisaccharide Me-beta-D-Glc6P-(1-->3)-alpha-D-Gal24N-[6-PC]-(1-->4)-alpha- D-galNAc and from the intra-ring NOE effects. 31P-NMR spectra point to a strong electrostatic or hydrogen-bonding interaction between the free 4-NH2 group on the Gal24N and the negatively charged diester phosphate group between adjacent pentasaccharide repeating-units [Ribitol-(5-->6)-beta-D-Glc6P]. Molecular modelling and MD simulation experiments confirmed that such an interaction was feasible with the Gal24N galactopyranose ring in the inverted B1.4 or skew 5S1 conformation.

Structural characterization of neutral glycosphingolipids from Trypanosoma cruzi

The major neutral glycosphingolipids from Trypanosoma cruzi ceramide mono- and dihexosides (CMH and CDH, respectively) were analysed after chromatographic purification using 1H 500 MHz nuclear magnetic resonance spectroscopy and fast atom bombardment mass spectrometry. The ceramide monohexoside fraction (CMH) contained both glucosyl- and galactosylceramides. After peracetylation, the CMH fraction was separated into 2 subfractions, CMH-COH and CMH-Cn, containing either hydroxy fatty acids or n-fatty acids. In the CMH-COH fraction glucose and galactose were present in a ratio of 2:1, whereas this ratio was 1:1 in the CMH-Cn fraction. The CDH fraction was identified as lactosylceramide with sphingosine as the long chain base and 16:0, 18:0, and 24:0, 24:4 fatty acids as major components.

Analysis of the seed oil of Heisteria silvanii (Olacaceae)—A rich source of a novel C18 acetylenic fatty acid

Besides some usual fatty acids (FA), two conjugated ene-yne acetylenic FA [trans-10-heptadecen-8-ynoic acid (pyrulic acid) (7.4%), and trans-11-octadecen-9-ynoic acid (ximenynic acid) (3.5%)], a novel ene-yne-ene acetylenic FA [cis-7, trans-11-octadecadiene-9-ynoic acid (heisteric acid) (22.6%)], and 9,10-epoxystearic acid (0.6%) could be identified in the seed oil of Heisteria silvanii (Olacaceae). Two further conjugated acetylenic FA [9,11-octadecadiynoic acid (0.1%) and 13-octadecene-9,11-diynoic acid (0.4%)] were identified tentatively by their mass spectra. The FA mixture has been analyzed by gas chromatography/mass spectrometry (GC/MS) of their methyl ester and 4,4-dimethyloxazoline derivatives. The structure of heisteric acid was elucidated after isolation via preparative silver ion thin-layer chromatography and by various spectroscopic methods [ultraviolet; infrared; 1H, 13C nuclear magnetic resonance (NMR); 1H−1H and 1H−13C correlation spectroscopy]. To determine the position of the conjugated ene-yne-ene system, the NMR spectra were also measured after addition of the lanthanide shift reagent Resolve-Al EuFODTM. Furthermore, the triyglyceride mixture was analyzed by high-temperature GC and high-temperature GC coupled with negative chemical ionization MS. A glass capillary column coated with a methoxy-terminated 50%-diphenyl-50%-dimethylpolysiloxane was used for the separation of the triacylglycerol (TAG) species. No evidence of decomposition of the TAG species containing conjugated ene-yne-ene FA was observed. Twenty-six species of the separated TAG were identified by means of their abundant quasi molecular ion [M−H]− and their corresponding carboxylate anions [RCOO]− of the fatty acids, respectively. The major molecular species of the TAG were found to be 16:0/18:1/18:1, 16:0/18:1/18:3 (heisteric acid), 17:2 (pyrulic acid)/18:1/18:1, 18:1/18:1/18:3 (heisteric acid). The TAG containing acetylenic FA showed an unexpected increase of the retention time in comparison to the TAG containing usual FA, thus making the prediction of the elution order of lipid samples containing acetylenic FA difficult.

The aqueous solution structure of the tetrasaccharide-ribitol repeat-unit from the lipoteichoic acid of Streptococcus pneumoniae strain R6 determined using a combination of NMR spectroscopy and computer calculations

High-resolution 1D- and 2D-correlation 1H NMR and 13C NMR, at 500 and 125 MHz, respectively, permitted assignment of the majority of the resonances in the per-N-acetylated, phosphorylated tetrasaccharide-ribitol repeat-unit, and in the complete polymer (n = 5 - 7) containing between five and seven repeating units attached to the deacylated lipid anchor, for the lipoteichoic acid from Streptococcus pneumoniae strain R6; the 31P resonances were also assigned. Comparison of the 31P spectra obtained for the per-N-acetylated oligosaccharide and for the oligosaccharide having the AATG 4-NH2 group still free, indicate a conformational difference brought about by interaction between the amino group and the neighboring phosphate group.

Triterpenes with a new 9-epi-cucurbitan skeleton from Senecio selloi

From the fresh aerial parts of Senecio selloi (Asteraceae), two diastereomeric triterpenes with a hitherto unknown skeleton have been isolated. The structure has been elucidated mainly by advanced NMR experiments, including inverse techniques, HMQC, HMBC, ROESY.

The structures of polysaccharides and glycolipids of Aspergillus fumigatus grown in the presence of human serum

A study was made of polysaccharides and glycosphingolipids isolated from Aspergillus fumigatus grown in media supplemented with human serum from healthy donors. Fractionation of Cetavlon-precipitated polysaccharides on Sephacryl S-400 gave rise to an excluded fraction (Fraction I) with molecular weight of >400 kDa and an included peak (Fraction II) with an average molecular weight of 30–80 kDa. Fraction I comprises about 5% of total polysaccharide and was identified as a glycogen-like molecule. Its structure was deduced from methylation data, treatment with amyloglucosidase, a red-brown coloration produced with an iodine solution and by 1H and 13C-NMR spectroscopy. It was previously suggested that higher amounts of glycogen-like polysaccharide (20%) were present in A. fumigatus grown in serum-free medium. Fraction II was identified as a galactomannan and was the main polysaccharide of A. fumigatus grown in serum-supplemented medium. Its structure was elucidated mainly by 13C-NMR spectroscopy combined with partial acetolysis and methylation analysis. The 13C-NMR spectrum of the galactomannan showed a much greater complexity in the β-d-galf and α-d-manp C-1 regions, than was evident for galactomannan from serum-free cultures previously described, reflecting differences in the glycosylation pattern, stimulated in serum-supplemented medium.No differences in A. fumigatus glycosphingolipid could be detected between serum-containing and serum-free growth conditions.Our results demonstrate that the change in polysaccharide structure is a more specific response to the altered growth conditions and not merely a symptom of more general changes.

Catecholamine-protein conjugates: isolation of 4-phenylphenoxazin-2-ones from oxidative coupling of N-acetyldopamine with aliphatic amino acids

4-Phenylphenoxazinones 4 were isolated after biomimetic oxidation, using diphenoloxidases of insect cuticle, mushroom tyrosinase, or after autoxidation of N-acetyldopamine (1) in the presence of β-alanine, β-alanine methyl ester or N-acetyl-L-lysine. They are formed presumably by addition of 2-aminoalkyl-5-alkylphenols 2 to the o-quinone of biphenyltetrol 3 which, in turn, arises from oxidative coupling of 1. The structures of 4 present the first examples for the assembly of reasonably stable intermediates in the rather complex process of chemical modifications of aliphatic amino acid residues by o-quinones.

The structural elucidation of the two positional isomers of a mono-glucopyranosyl mono-acyl glycerol derivative from Cystobacter fuscus (Myxobacterales) ☆

The structures of two glycolipids produced by Cystobacter fuscus Cb 685 have been determined by 1H-NMR spectroscopy including 2D-techniques, as the positional isomers 1- and 2-isopentadecanoyl-3-beta-D-glucopyranosyl-X-glycerol. The chain length of the fatty acid residue determined by 13C-NMR spectroscopy has been confirmed by negative ion FAB-mass spectrometry.