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M Bernabé

Publications and source records attributed to M Bernabé.

At least 19 recordsLinked to original sources
PubMed2005

alpha-galf I -->6-alpha-mannopyranoside side chains in Paracoccidioides brasiliensis cell wall are shared by members of the Onygenales, but not by galactomannans of other fungal genera.

The water-soluble polysaccharide fraction of the cell wall alkali extract (F1SS) from the mycelial phase of the dimorphic fungus Paracoccidioides brasiliensis is compared with F1SS polysaccharides obtained from the Onygenalean mycelial fungi Ascocalvatia alveolata, Onygena equina and Aphanoascus terreus. These polymers were exclusively composed of mannose and galactose. Data from methylation and NMR analyses reveal that F1SS polysaccharides from the four fungi contain the same residues although in different proportions: [-->2,6)-alpha-D-Manp-(1 -->]; [2)-alpha-D-Manp-(1 -->]; [ -->6)-alpha-D-Manp-(1 -->]; and [alpha-D-Galf-(1 -->]. In P. brasiliensis, the repeating unit of the polysaccharide consists of a backbone of [(1 -->6)-alpha-D-Manp] substituted at the 0-2 position by the disaccharide [alpha-D-Galf-(1 -->6)-alpha-D-Manp-(1 -->], while the remaining 0-2 positions are substituted by single residues of mannose or short chains of (1 -->2)-mannose. The other species had a lower proportion of galactofuranose-containing side chains and higher proportion of mannose-containing side chains. The similarities found among the F1SS polysaccharides from P. brasiliensis and the Onygenalean A. alveolata, A. terreus and O. equina, reveal the close relatedness of all these fungi, show differences with polysaccharides from other fungal genera and agree with the molecular evidence provided in the scientific literature for the placement of P. brasiliensis within the Onygenales.

Carbohydrate Sequence↗
PubMed2000

Heterogeneity of the genus Myrothecium as revealed by cell wall polysaccharides.

The polysaccharides obtained from the alkali-extractable, water-soluble fraction (F1SS) from the cell wall of Myrothecium verrucaria and Myrothecium atroviride were shown to be composed of beta-(1-->6)-galactofuranose fully substituted at O-2 by terminal residues of alpha-glucopyranose and alpha-glucuronic acid. Glucuronic acid was substituted at O-4 by glucopyranose in the Myrothecium species M. inundatum, M. setiramosum, M. prestonii, M. tongaense and M. roridum. The acidic polysaccharides from Phaeostilbella atra (=Myrothecium atrum) and Myrothecium gramineum lacked the backbone of 2,6 di-O-substituted galactofuranose and contained a high amount of O-5-substituted beta-galactofuranose. The structures of the polysaccharides isolated from Myrothecium cinctum and Myrothecium penicilloides were unrelated to each other and to the polysaccharides from the other species analysed. The usefulness of these polysaccharides as a characteristic for delimitation of the genus Myrothecium is discussed.

Ascomycota↗
PubMed2000

Four new triterpenoid saponins from the roots of Bupleurum rigidum.

Five triterpenoid saponins [buddlejasaponin IV, sandrosaponins VII (1), VIII (2), IX (3), and X (4)] were isolated from an n-BuOH extract of the roots of Bupleurum rigidum. Sandrosaponins VII-X (1-4) are new compounds, and their structures were established by 1D and 2D NMR techniques, FABMS, and chemical methods.

Apiaceae↗
PubMed2000

Purification and structure of neurostatin, an inhibitor of astrocyte division of mammalian brain.

Neurostatin was originally described as an inhibitor of astroblast and astrocytoma division present in rat brain extracts and immunologically related to the sugar moiety of epidermal growth factor receptor and to blood group antigens. It was purified recently from mammalian brain extracts and characterized as a glycosphingolipid, but its precise structure remained unknown. Neurostatin has now been purified to apparent homogeneity from ganglioside extracts of rat, bovine, and porcine brain. It is cytostatic for astroblasts, C6 glioma cells, and various human astrocytomas grades III and IV, with IC(50) values ranging from 250 to 450 nM, but does not affect the division of primary or transformed fibroblasts up to concentrations >4 microM. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry of purified pig neurostatin showed a molecular ion of 1, 905 Da and ions of 1,863 and 1,934 Da, compatible with a disialoganglioside. Mono- and bidimensional NMR spectra, together with biochemical studies, suggest that neurostatin may be the 9-O-monoacetyl ester of GD1b.

3T3 Cells↗
PubMed1999

Lipase-catalyzed regioselective acylation of sucrose in two-solvent mixtures.

The enzymatic synthesis of 6-O-lauroylsucrose and 6-O-palmitoylsucrose was performed by transesterification of sucrose with the corresponding vinyl esters in a medium constituted by two solvents. More specifically, the acylation was carried out in 2-methyl-2-butanol (tert-amyl alcohol) containing a low percentage (not higher than 20%) of dimethyl sulfoxide. Several lipases were able to catalyze the transesterification, but that from Humicola lanuginosa (adsorbed on diatomaceous earth) was particularly useful. We optimized the synthesis of 6-O-lauroylsucrose varying the percentage and nature of the cosolvent, the molar ratio sucrose/vinyl laurate, the nature of bulk solvent and the enzyme content. Under the best conditions (2-methyl-2-butanol/DMSO 4:1 v/v), a sucrose conversion of 70% to 6-O-lauroylsucrose was achieved in 24 h using 50 mg biocatalyst/mL. As a side process, a low percentage (<5% in 24 h) of the initial sucrose is converted into the diesters 6,1'-di-O-lauroylsucrose and 6,6'-di-O-lauroylsucrose. The above methodology was also extended to the synthesis of 6-O-palmitoylsucrose. The acylation process was even faster, giving rise to an 80% conversion to monoester in 48 h using 25 mg biocatalyst/mL. This study shows that the use of two-solvent mixtures may become a feasible alternative for the synthesis of sucrose esters, allowing to exploit the catalytic potential of lipases.

Acylation↗
PubMed1999

Galactomannans from the cell walls of species of Paecilomyces sect. Paecilomyces and their teleomorphs as immunotaxonomic markers.

An alkali-extractable and water-soluble fraction (F1S) was obtained from cell walls of Paecilomyces variotii and species of the related genera Talaromyces, Byssochlamys and Thermoascus. The structure of the main polysaccharide of these fractions was studied and found to consist of a core of (1 --> 6)-alpha-mannopyranose partially substituted at 0.2 by chains of galactofuranose and shorter chains of mannopyranose. The differences in the regularity of the branching points and the length of the galactofuranose side chains are useful to distinguish between species. These differences were detected by immunological methods, since highly specific polyclonal antibodies were raised against these polysaccharides. Mycelium of P. variotii CBS 990.73A was stained by indirect immunofluorescence. The polysaccharides studied in this work differ from the one described for species from section Isarioidea, and this is another indication of the heterogeneity of the genus Paecilomyces.

Animals↗
PubMed1998

A saponin from the roots of Gypsophila bermejoi.

A new saponin was isolated from the methanol extract of the roots of Gypsophila bermejoi and identified by a combination of chemical degradation and spectral methods, which included negative FABMS and extensive 1D and 2D-NMR analysis (DQCOSY, TOCSY, ROESY, HMQC and HMBC), as gypsogenin 28-O-beta-D-glucopyranosyl(1-->2)-[beta-D-glucopyranosyl (1-->6)]-beta-D-glucopyranoside.

Carbohydrate Sequence↗
PubMed1998

Two new sulfated saponins from the roots of Gypsophila bermejoi.

Two new sulfated saponins (1 and 2) were isolated from a butanol-soluble extract of the roots of Gypsophila bermejoi and were identified by a combination of chemical degradation and spectral methods as the 3beta-sulfate ester of gypsogenin 28-O-beta-D-glucopyranosyl(1-->2)-[beta-D-glucopyranosyl(1-->6)]-beta -D-glucopyranoside (1), and the 23-sulfate ester of hederagenin 28-O-beta-D-glucopyranosyl(1-->2)-[beta-D-glucopyranosyl(1-->6)]-beta -D-glucopyranoside (2), respectively. Plants of the genus Gypsophila (Caryophyllaceae) are important industrially because of the capacity of their saponin constituents to behave like natural detergents.1 Saponins from this genus are based on oleanolic acid (3-hydroxyolean-12-en-28-oic acid), with gypsogenin being the main pentacyclic triterpenoidal aglycon found. In general, this aglycon is substituted at the C-3 hydroxyl and/or C-28 carboxylic acid groups by saccharide units.2

Carbohydrate Sequence↗
PubMed1997

Studies on the structure and the solution conformation of an acidic extracellular polysaccharide isolated from Bradyrhizobium.

The structure of an acidic extracellular polysaccharide isolated from Bradyrhizobium (Chamaecytisus proliferus) has been elucidated by hydrolysis, methylation analysis, and 1D and 2D 1H- and 13C-NMR spectroscopy of the complete polysaccharide. The NMR spectrum showed that microheterogeneity was present due to the minor existence of a variety of O-acetyl groups. Thus, a deacetylated sample was prepared by alkaline treatment which was then fully analysed. The deacetylated polysaccharide has the following sequence: -->3)-[alpha-D-Galp-(1-->6)-alpha-D-Glcp-(1-->3)-beta-D-Glcp-(1-->3)- alpha-D-GalpA-(1-->3)-alpha-D-Manp-(1--> The sample is partially O-methylated at position 4 of the alpha-D-Galp-(1-->6) unit. In addition, the same moiety of the native sample is also partially and heterogeneously O-acetylated. The conformational features of the deacetylated sample have been evaluated by molecular mechanics and dynamics calculations and NOE spectroscopy. The results indicate that the polysaccharide may adopt a variety of three dimensional shapes, and that there is a fair agreement between the NMR-derived distances and those provided by the calculations

Carbohydrate Conformation↗
PubMed1997

Solution conformation and dynamics of an extracellular polysaccharide isolated from Bradyrhyzobium as deduced from 1H-NMR off resonance ROESY and 13C-NMR relaxation measurements.

The conformational and dynamical features of an extracellular branched deacetylated polysaccharide isolated from Bradyrhizobium (Chamaecytisus proliferus) have been investigated by homo and heteronuclear NMR methods. 1H-NMR cross relaxation rates have been obtained for this polysaccharide through regular NOESY and ROESY spectra as well as by modern off resonance ROESY techniques. Local proton-proton correlation times as well as interproton distances have been obtained. 13C-NMR relaxation parameters (T1, T2, NOE) have also been measured at two different magnetic fields and interpreted using different approximations based on the Lipari and Szabo model free approach. The analysis of the data indicates the existence of important flexibility for the different linkages of the polysaccharide. Motions in the range of several ns contribute to the relaxation of the macromolecule, although faster internal motions in the 600-800 ps time scales are also present. These time scales indicate that segmental motions as well as internal motions around the glycosidic linkages are the major sources of relaxation for this molecule at 299 K.

Carbohydrate Conformation↗
PubMed1997

Structure of complex cell wall polysaccharides isolated from Trichoderma and Hypocrea species.

The structure of fungal polysaccharides isolated from the cell wall of Trichoderma reesei, T. koningii, and Hypocrea psychrophila, have been investigated by means of chemical analyses and 1D and 2D NMR spectroscopy. The polysaccharides have an irregular structure, idealized as follows: [formula: see text] The proportions of the different side chains vary from a species to another, being n above some three times larger in H. psychrophila than in T. reesei or T. koningii.

Carbohydrate Conformation↗
PubMed1997

Structural elucidation of acidic fungal polysaccharides isolated from the cell-wall of genera Cylindrocladium and Calonectria.

The structure of acidic fungal polysaccharides, isolated from the cell wall of Cylindrocladium and Calonectria species, has been investigated by chemical analysis, methylation and reductive cleavage analyses, and 1D and 2D 1H and 13C NMR spectroscopy. The polysaccharides have an idealized repeating unit: [formula: see text] linked to a small mannan core (< 5%), with n = 3 for Cylindrocladium penicilloides and C. quinqueseptatum and n = 5 for Calonectria theae, C. crotalariae, and C. colhounii.

Carbohydrate Sequence↗
PubMed1996

An antigenic water-soluble glucogalactomannan extracted from cell walls of Paecilomyces fumosoroseus and Paecilomyces farinosus.

The water-soluble fraction (F1S) obtained after solubilizing in alkali the cell walls of four strains of Paecilomyces fumosoroseus and two of Paecilomyces farinosus amounted to 8.3-14.5% of the dry cell wall material. Two polysaccharides, F1S-A (13-20%) and F1S-B (57-68%) were separated from F1S by gel permeation through Sepharose CL-6B. 1H and 13C NMR spectra of F1S-B were recorded and showed analogous structural features in the six isolates of the two species. The fractions isolated from P. fumosoroseus strain CBS 375.70 were subjected to structural analysis and shown to be a (1-->4)-alpha-glucan (F1S-A) and a branched (1-->6)-mannan with terminal residues of beta-galactopyranose (F1S-B). Polyclonal antibodies against the latter polysaccharide were obtained (titre 1/8000). These antibodies reacted specifically with the F1S-B polysaccharides obtained from the four strains of P. fumosoroseus and the two strains of P. farinosus, but they did not react with similar fractions from other species of the same or related genera. The antibodies specifically stained P. fumosoroseus hyphae in indirect immunofluorescence tests.

Antigens, Fungal↗