SUMMARY: Spheroplasts of were regenerated in [C]glucose and buffered magnesium sulphate (0.1 -Tris/HC1; 0.5 -MgSO, pH 7.2) at 35°C. Uptake of glucose by spheroplasts was faster than that by intact yeast cells. After 6 h, 65% of the glucose taken up by the yeast appeared as CO and 30% was incorporated into the cellular material. With spheroplasts, 55% of the glucose taken up was expired as CO, 25% was excreted into the medium as other metabolites and 20% was incorporated into the cells. The regenerating spheroplasts excreted C-labelled carbohydrates into the medium which were fractionated on a Sephadex G-15 column. Acid hydrolysis of the low molecular-weight fraction yielded the following sugars: mannose (75.7%), fucose (3.8%), arabinose (3%), galactose (2.1%) and an unidentified monosaccharide (14%). Spheroplasts did not incorporate mannoprotein into the regenerated wall. The wall carbohydrate from regenerated spheroplasts was fractionated on the basis of solubility in sodium hydroxide. The alkali-insoluble fraction was analysed by sequential enzyme hydrolysis; 40% of the incorporated counts were associated with β(1 → 3)-linked glucan and 50% with a mixed glucan comprising β(1 → 3)- and β(1 → 6)-linkages and chitin.


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