B-LYMPHOCYTE-STIMULATING POLYSACCHARIDE FROM MUSHROOM PHELLINUS-LINTEUS

Hot water extract prepared from the mycelial culture of mushroom Phell inus linteus stimulated polyclonal antibody production in an in vitro culture system. The active fraction PLP was purified from the extract ca, 1030-fold by ethanol precipitation followed by DEAE-cellulose and gel permeation chromatography. PLP contained 13.2% (w/w) peptide and 8 2.5% (w/w) carbohydrate. About 6.8% (w/w) of the total carbohydrate wa s uronic acid. The molecular weight distribution of PLP was found to b e nearly homogeneous (153 kDa) in gel permeation HPLC analysis. Neutra l sugar composition analysis revealed Ara (7.0%), Xyl (3.7%), Glc (21. 1%), Gal(24.1%) and Man (44.2%). Uronic acid was identified as a glucu ronic acid by gas chromatography. Ten amino acids were detected and As p and Glu were the major components, In our assay system, the half-max imal concentration of PLP for B-lymphocyte stimulation was ca, 3 mu g/ ml. Partial acid hydrolysis as well as sodium periodate treatment of P LP decreased the activity significantly, suggesting that both the full molecular size and the sugar moiety were essential. However, proteina se K treatment for up to 48 h did not affect the activity.