A study of the phosphate linkages in phosphomannan in cell walls of Saccharomyces cerevisiae

Abstract

1. The phosphomannan of Saccharomyces cerevisiae was released by Pronase digestion of cell walls and isolated by chromatography on DEAE-cellulose or by precipitation with borate–Cetavlon solutions. Mannose and phosphorus were present in the molar ratio 18:1 and the phosphate groups were in the diester form. 2. Hydrolysis with acid gave mannose 6-phosphate. Under mild acid conditions (autohydrolysis) the phosphate groups were converted into the monoester form, mannose was released and the molecular size of the phosphomannan was substantially decreased. 3. Hydrolysis with alkali also gave a monoester phosphate and a similar decrease in molecular weight. Under mild alkaline conditions the serine and threonine content of the phosphomannan was decreased by about 80%. The phosphate content was not altered. 4. Treatment with 40% (v/v) HF removed 70% of the phosphorus from the phosphomannan with no detectable decrease in molecular weight. 5. Periodate oxidation gave an oxophosphomannan from which 80% of the phosphorus was eliminated under mild alkaline conditions. 6. The properties of the phosphomannan are consistent with a structure in which the phosphate groups are located on the outside of the molecule and link C-1 of a terminal mannose unit with C-6 of another mannose unit, which is in turn attached to the polysaccharide backbone of the molecule. 7. The implications of this structure are discussed in relation to flocculation.