Polysaccharides α-D-galactan (GAL-Am) and β-D-glucan (GLC-Am) were obtained from Amanita muscaria fruiting bodies. They were purified using different methodologies, such as Fehling precipitation (for both fractions), freeze-thawing process and ultrafiltration (for GLC-Am). Results showed that the GAL-Am has (1→6)-linked Galp main chain branched at O-2 by terminal Galp units and has not been previously reported. Besides, GLC-Am has (1→3)-linked Glcp in the main chain, substituted at O-6 by (1→6)-linked β-Glcp units. Both are water-soluble, with 9.0 × 103 g/moL and 1.3 × 105 g/moL, respectively. GAL-Am and GLC-Am presented a selective proliferation reduction against B16-F10 melanoma cell line, not affecting non tumoral BALB/3T3 fibroblast cell line. Furthermore, both fractions reduced clonogenic capacity of melanoma cell line over an extended period of time. These results were obtained without modulations in B16-F10 cell adhesion, reinforcing the biological activities towards cell proliferation impairment and eliciting these polysaccharides as promising compounds to further exploration of their antimelanoma properties.

polysaccharides, chemical structure, β-glucan, β-D-glucan, α-galactan, A. muscaria, antimelanoma properties

13C NMR data:
Linkage	Residue	C1	C2	C3	C4	C5	C6
6	aDGalp	98.1	69.6	68.4	69.5	68.8	66.7
2	aDGalp	95.7	69.6	69.5	69.5	71.0	61.2
	aDGalp	98.0	75.1	66.9	66.2	68.8	66.7


1H NMR data:
Linkage	Residue	H1	H2	H3	H4	H5	H6
6	aDGalp	4.98	3.88	3.85	4.03	4.17	3.71-3.91
2	aDGalp	5.15	3.88	3.94	4.03	4.18	3.76-3.76
	aDGalp	5.02	3.97	3.98	4.25	4.17	3.71-3.91


1H/13C HSQC data:
Linkage	Residue	C1/H1	C2/H2	C3/H3	C4/H4	C5/H5	C6/H6
6	aDGalp	98.1/4.98	69.6/3.88	68.4/3.85	69.5/4.03	68.8/4.17	66.7/3.71-3.91
2	aDGalp	95.7/5.15	69.6/3.88	69.5/3.94	69.5/4.03	71.0/4.18	61.2/3.76-3.76
	aDGalp	98.0/5.02	75.1/3.97	66.9/3.98	66.2/4.25	68.8/4.17	66.7/3.71-3.91

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

[*]OC[C@H]1O[C@H](OC[C@H]2O[C@H]([*])[C@H](O[C@H]3O[C@H](CO)[C@H](O)[C@H](O)[C@H]3O)[C@@H](O)[C@H]2O)[C@H](O)[C@@H](O)[C@H]1O

486.423 g/mol (C₁₈H₃₀R₂O₁₅, R = next and previous repeats, not counted in mol weight)

Polysaccharides α-D-galactan (GAL-Am) and β-D-glucan (GLC-Am) were obtained from Amanita muscaria fruiting bodies. They were purified using different methodologies, such as Fehling precipitation (for both fractions), freeze-thawing process and ultrafiltration (for GLC-Am). Results showed that the GAL-Am has (1→6)-linked Galp main chain branched at O-2 by terminal Galp units and has not been previously reported. Besides, GLC-Am has (1→3)-linked Glcp in the main chain, substituted at O-6 by (1→6)-linked β-Glcp units. Both are water-soluble, with 9.0 × 103 g/moL and 1.3 × 105 g/moL, respectively. GAL-Am and GLC-Am presented a selective proliferation reduction against B16-F10 melanoma cell line, not affecting non tumoral BALB/3T3 fibroblast cell line. Furthermore, both fractions reduced clonogenic capacity of melanoma cell line over an extended period of time. These results were obtained without modulations in B16-F10 cell adhesion, reinforcing the biological activities towards cell proliferation impairment and eliciting these polysaccharides as promising compounds to further exploration of their antimelanoma properties.

polysaccharides, chemical structure, β-glucan, β-D-glucan, α-galactan, A. muscaria, antimelanoma properties

13C NMR data:
Linkage	Residue	C1	C2	C3	C4	C5	C6
3,3	bDGlcp	103.2	72.8	86.3-86.7	68.7	76.4	60.9
3,6	bDGlcp	103.2	73.7	76.7	70.2	76.4	60.9
3	bDGlcp	103.2	72.8	86.1	68.7	74.8	68.5
	bDGlcp


1H NMR data:
Linkage	Residue	H1	H2	H3	H4	H5	H6
3,3	bDGlcp	4.55	3.35	3.50	3.29	3.29	3.50-3.72
3,6	bDGlcp	4.27	3.09	3.14	3.14	3.29	3.50-3.72
3	bDGlcp	4.55	3.35	3.51	3.29	3.54	3.58-4.11
	bDGlcp


1H/13C HSQC data:
Linkage	Residue	C1/H1	C2/H2	C3/H3	C4/H4	C5/H5	C6/H6
3,3	bDGlcp	103.2/4.55	72.8/3.35	86.3-86.7/3.50	68.7/3.29	76.4/3.29	60.9/3.50-3.72
3,6	bDGlcp	103.2/4.27	73.7/3.09	76.7/3.14	70.2/3.14	76.4/3.29	60.9/3.50-3.72
3	bDGlcp	103.2/4.55	72.8/3.35	86.1/3.51	68.7/3.29	74.8/3.54	68.5/3.58-4.11
	bDGlcp

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

[*]O[C@H]1[C@H](O)[C@@H](CO)O[C@@H](O[C@@H]2[C@@H](O)[C@H](O[C@@H]3[C@@H](O)[C@H]([*])O[C@H](CO)[C@H]3O)O[C@H](CO[C@@H]3O[C@H](CO)[C@@H](O)[C@H](O)[C@H]3O)[C@H]2O)[C@@H]1O

648.564 g/mol (C₂₄H₄₀R₂O₂₀, R = next and previous repeats, not counted in mol weight)

Polysaccharides α-D-galactan (GAL-Am) and β-D-glucan (GLC-Am) were obtained from Amanita muscaria fruiting bodies. They were purified using different methodologies, such as Fehling precipitation (for both fractions), freeze-thawing process and ultrafiltration (for GLC-Am). Results showed that the GAL-Am has (1→6)-linked Galp main chain branched at O-2 by terminal Galp units and has not been previously reported. Besides, GLC-Am has (1→3)-linked Glcp in the main chain, substituted at O-6 by (1→6)-linked β-Glcp units. Both are water-soluble, with 9.0 × 103 g/moL and 1.3 × 105 g/moL, respectively. GAL-Am and GLC-Am presented a selective proliferation reduction against B16-F10 melanoma cell line, not affecting non tumoral BALB/3T3 fibroblast cell line. Furthermore, both fractions reduced clonogenic capacity of melanoma cell line over an extended period of time. These results were obtained without modulations in B16-F10 cell adhesion, reinforcing the biological activities towards cell proliferation impairment and eliciting these polysaccharides as promising compounds to further exploration of their antimelanoma properties.

polysaccharides, chemical structure, β-glucan, β-D-glucan, α-galactan, A. muscaria, antimelanoma properties

13C NMR data:
Linkage	Residue	C1	C2	C3	C4	C5	C6
	bDGlcp	103.0	72.9	86.2	68.4	76.4	60.9


1H NMR data:
Linkage	Residue	H1	H2	H3	H4	H5	H6
	bDGlcp	4.53	3.31	3.49	3.25	3.27	3.48-3.71


1H/13C HSQC data:
Linkage	Residue	C1/H1	C2/H2	C3/H3	C4/H4	C5/H5	C6/H6
	bDGlcp	103.0/4.53	72.9/3.31	86.2/3.49	68.4/3.25	76.4/3.27	60.9/3.48-3.71

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

[*]O[C@@H]1[C@@H](O)[C@H]([*])O[C@H](CO)[C@H]1O

162.141 g/mol (C₆H₁₀R₂O₅, R = next and previous repeats, not counted in mol weight)