Mengxia Yan

Chemical characteristic of an anticoagulant-active sulfated polysaccharide from Enteromorpha clathrata

A sulfated polysaccharide FEP from marine green alga Enteromorpha clathrata was extracted with hot water and further purified by ion-exchange and size-exclusion chromatography. Results of chemical and spectroscopic analyses showed that FEP was a high arabinose-containing sulfated polysaccharide with sulfate ester of 31.0%, and its average molecular weight was about 511kDa. The backbone of FEP was mainly composed of (1→4)-linked β-L-arabinopyranose residues with partially sulfate groups at the C-3 position. In vitro anticoagulant assay indicated that FEP effectively prolonged the activated partial thromboplastin time and thrombin time. The investigation demonstrated that FEP was a novel sulfated polysaccharide with different chemical characteristics from other sulfated polysaccharides from marine algae, and could be a potential source of anticoagulant.

Structural characterization and anticoagulant activity of a sulfated polysaccharide from the green alga Codium divaricatum

A sulfated polysaccharide, designated CP2-1, was isolated from the green alga Codium divaricatum by water extraction and purified by anion-exchange and size-exclusion chromatography. CP2-1 is a galactan which is highly sulfated and substituted with pyruvic acid ketals. On the basis of chemical and spectroscopic analyses, the backbone of CP2-1 was mainly composed of (1→3)-β-d-galactopyranose residues, branched by single (1→)-β-d-galactopyranose units attached to the main chain at C-4 positions. The degree of branching was estimated to be about 12.2%. Sulfate groups were at C-4 of (1→3)-β-d-galactopyranose and C-6 of non-reducing terminal galactose residues. In addition, the ketals of pyruvic acid were found at 3,4- of non-reducing terminal galactose residues forming a five-membered ring. CP2-1 possessed a high anticoagulant activity as assessed by the activated partial thromboplastin time and thrombin time assays. The investigation demonstrated that CP2-1 was an anticoagulant-active sulfated polysaccharide distinguishing from other sulfated polysaccharides from marine green algae.

Structural elucidation of an extracellular polysaccharide produced by the marine fungus Aspergillus versicolor

A homogenous extracellular polysaccharide, designated AWP, was isolated from the fermented liquid of the marine fungus Aspergillus versicolor from the coral Cladiella sp. and purified by anion-exchange and size-exclusion chromatography (SEC). Chemical and spectroscopic analyses, including one- and two-dimensional nuclear magnetic resonance (1D and 2D NMR) spectroscopy showed that AWP consisted of glucose and mannose in a molar ratio of 8.6:1.0, and its average molecular weight was estimated to be 500kDa. AWP is a slightly branched extracellular polysaccharide. The backbone of AWP is mainly composed of (1→6)-linked α-D-glucopyranose residues, slightly branched by single α-d-mannopyranose units attached to the main chain at C-3 positions of the glucan backbone. The investigation demonstrated that AWP is a novel extracellular polysaccharide different from those of other marine microorganisms.

Galactomannan with novel structure produced by the coral endophytic fungus Aspergillus ochraceus

The homogeneous extracellular polysaccharide, AW1, was obtained from the fermented broth of the fungus Aspergillus ochraceus derived from coral Dichotella gemmacea. AW1 was a galactomannan with a molar ratio of mannose and galactose of 2.16:1.00 and a molecular weight of about 29.0kDa. The structure of AW1 was investigated by chemical and spectroscopic methods, including methylation analysis, one- and two-dimensional nuclear magnetic resonance (1D, 2D NMR) and electrospray mass spectrometry with collision-induced dissociation (ES-CID MS/MS) spectroscopic analyses. The results showed that the backbone of AW1 consisted of (1⟶2)-linked α-d-mannopyranose residues. The mannopyranose residues in the backbone were substituted at C-6 by the (1⟶)-linked α-d-mannopyranose units and (1⟶5)-linked β-d-galactofuranose oligosaccharides with different degrees of polymerization. The investigation demonstrated that AW1 was a novel galactomannan with different structural characteristics from other fungal galactomannans, and could be a potential resource of the (1⟶5)-linked β-d-galactofuranose oligosaccharides.

Extracellular polysaccharide with novel structure and antioxidant property produced by the deep-sea fungus Aspergillus versicolor N2bc.

An extracellular polysaccharide, N1, was obtained from the culture medium of the deep-sea fungus Aspergillus versicolor N2bc by a combination of ethanol precipitation, ion-exchange and gel filtration chromatography. N1 was a mannoglucogalactan with molecular weight of about 20.5kDa. Results of chemical and spectroscopic analyses, including Fourier-transform infrared, one- and two-dimensional nuclear magnetic resonance spectroscopy showed that the main chain of N1 consisted of →2)-α-d-Glcp-(1→, →2)-β-d-Glcp-(1→ and →6)-β-d-Manp-(1→ units, substituted at C-6 position of →2)-α-d-Glcp-(1→ units. The branches were composed of galactofuranose-oligosaccharides built up of →5)-β-d-Galf-(1→, →6)-β-d-Galf-(1→ and terminal β-d-Galf units. At an average, there were two branching points for every five sugar residues in the backbone. N1 possessed a high in vitro antioxidant activity as evaluated by scavenging assays involving superoxide, 1,1-diphenyl-2-picrylhydrazyl, hydroxyl radicals and reducing power. The investigation revealed that N1 was a novel antioxidant polysaccharide differing from previously described extracellular polysaccharides and could be a potential antioxidant.

Structural elucidation of the exopolysaccharide produced by the mangrove fungus Penicillium solitum.

A water soluble extracellular polysaccharide, designated GW-12, was obtained from the liquid culture broth of the mangrove fungus Penicillium solitum by ethanol precipitation, anion-exchange and size-exclusion chromatography. Reversed-phase high performance liquid chromatography analysis showed that GW-12 mainly consisted of d-mannose, and its molecular weight was estimated to be about 11.3 kDa determined by high performance gel permeation chromatography. On the basis of chemical and spectroscopic analyses, including methylation analysis and nuclear magnetic resonance (NMR) spectroscopy, the structure of GW-12 may be represented as a mannan with branches. The main chain of GW-12 was composed of (1 → 2)-linked α-D-mannopyranose and (1 → 6)-linked α-D-mannopyranose residues, branched by single α-d-mannopyranose units attached to the main chain at C-6 positions of (1 → 2)-linked α-D-mannopyranose residues. There was three branch points for every seven sugar residues in the backbone.

Preparation and structural elucidation of a glucomannogalactan from marine fungus Penicillium commune.

The coral-associated fungus Penicillium commune produces an extracellular polysaccharide, FP2-1, when grown in potato dextrose-agar medium. FP2-1 was isolated from the fermented broth using anion-exchange and size-exclusion chromatography, and its structure was elucidated by chemical and spectroscopic analyses, including detailed nuclear magnetic resonance spectroscopy. The results showed that FP2-1 was a glucomannogalactan with a molar ratio of galactose, mannose and glucose of 3.9:1.9:1.0. Structure of FP2-1 may be represented, at an average, as a backbone of (1→2)-linked α-mannopyranose with the every second residue substituted at position 6 by a pentasaccharide branch. The branches consist of four (1→6)-linked β-galactofuranose residues with terminal α-glucopyranose residue attached to the last galactofuranose residue at position 2. FP2-1 was a novel galactofuranose-containing extracellular polysaccharide differing from previously described extracellular polysaccharides.

Sequence analysis of the sulfated rhamno-oligosaccharides derived from a sulfated rhamnan

Three sulfated rhamno-oligosaccharides, designated O1, O2 and O3, were obtained by mild acid hydrolysis of the sulfated rhamnan and purified by gel-permeation chromatography. On the basis of one- and two-dimensional nuclear magnetic resonance (1D, 2D NMR) spectroscopic analyses, the oligosaccharide O1 was characterized to be α-L-Rhap-(2SO4)-(1→3)-α-L-Rhap. The fragmentation pattern of the homogeneous disaccharide in the product ion spectra was recognized by negative-ion electrospray tandem mass spectrometry with collision-induced dissociation (ES-CID MS/MS). With the principles established, the sequences of the oligosaccharides O2 and O3 were deduced to be α-L-Rhap-(2SO4)-(1→3)-α-L-Rhap-(1→3)-α-L-Rhap, and α-L-Rhap-(2SO4)-(1→3)-α-L-Rhap-(1→3)-α-L-Rhap-(1→3)-α-L-Rhap (2SO(4)), respectively. The investigation demonstrated that the sulfated rhamnan-derived oligosaccharides were novel sulfated oligosaccharides different from those of other polysaccharides-degraded from algae, and it could be possible to determine the sequence of the sulfated rhamno-oligosaccharides directly from the glycosidic cleavage fragmentation in the product ion spectra.