Isolation and identification of poly-α-(1→4)-linked 3-O-methyl-d-mannopyranose from a hot-water extract of Mycobacterium vaccae

Abstract A polysaccharide around 3.6 kDa has been identified as the major carbohydrate moiety of a antineoplastic protein–polysaccharide complex (PS4A) obtained by boiling intact cells of Mycobacterium vaccae in water. 1 H and 13 C NMR spectra of this polysaccharide suggested it was a highly homogeneous polymer composed substantially of one monomer, probably an α-linked O-methylated mannose. Comparison of the COSY spectra of the original and acetylated polymer indicated that the glycosidic linkage and the methyl ether were interchangeable, at O-3 and O-4. Further study demonstrated that the benzyolated hydrolysate of the polymer was 1,2,4,6-tetra- O -benzoyl-3- O -methyl-β-mannopyranose. The hydrolysate was 3- O -methyl-α,β-mannopyranose and the polymer was therefore poly-α-(1→4)-linked 3- O -methyl- d -mannopyranose. This conclusion was further confirmed with an authentic sample of the monomer, which had spectral data identical to those of the hydrolyzate and co-eluted from an ion-exchange HPLC with the major sugar in the hydrolysate.