Further Studies on Purification and Antigenic Activity of Saccharomyces cerevisiae Glucan

概要

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Acetone dried cells of Saccharomyces cerevisiae were treated with 45% phenol and further twice with hot water for 3 hr. at 135 in an autoclave to remove mannan, and then the resulting cell residue was extracted with 3% NaOH under N2 gas for 40 min. at 70-80. From the extracts in dialyzed bag, alkali-soluble glucan precipitated as gelatinous solid. The polysaccharide was autoclaved in 0.5NHOAc at 130 for 30 min., and thus obtained water-soluble glucan was fractionated with DEAE-Sephadex column chromatography, borate form, (GI and II). By analysis of sugar component of the eluted fractions, it was found that GI comprise a trace of mannose in addition to glucose and GII shows the presence of glucose as only sugar. Nitrogen and phosphorus were not detected in glucan fractions. Glucan antigens showed extremely weak precipitin activity against whole-cell antiserum of S. cerevisiae which was absorbed with purified mannan, and it did not showed the possibility to render immune for rabbits at whole cells of the organism. On the other hand, when cell-wall antiserum and the same antiserum absorbed with mannan were used, GII fraction showed high titer of 1 : 128 × 103 against both sera, and the results indicated that antibody responsible for precipitation of glucan is formed in rabbit serum by use of the autolyzed cells.
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