Chemical and enzymic analyses of Pichia polymorpha cell walls

Conventional techniques of chemical analysis have shown that the cell walls of the yeast Pichia polymorpha, at early stationary growth phase, consisted of carbohydrate (about 85%), protein (8%), and lipid (7%). Glucose and mannose were the only neutral sugars and glucosamine the sole amino sugar present among the cell wall components. Paper and gas-liquid chromatographies of acid hydrolysates of purified cell walls and cell wall fractions proved that mannan and alkali-insoluble glucans, in that order, were the major polysaccharide components, accounting for 83% of the total carbohydrate content. The isolation of an alkali-soluble glucan, non-precipitable with Fehling's solution, has been achieved. Treatments of whole cell walls and their fractions with purified cell wall lytic enzymes have shown the presence of both 1,3- and 1,6-beta-D-linkages in all the glucan fractions. 1,3-alpha-Glucan was not detected. Mannan-glucan complexes have been found containing about 50% each of mannose and glucose. All polysaccharides exhibited different turnover rates when cells were grown in the presence of D-[U-14C]glucose. The morphogenetic implications of these results are discussed.