The optimal condition for autolysis of spent brewer’s yeast from molasses fermentation at yeast suspension of 15% (w/w) solid content and pH 5.0 was 50 ^oC for 24 hours. This condition resulted in high autolysed yeast cells and protein autolysate. Autolysed cell was separated and soluble protein was extracted out with hot water (121 ^oC) for 1, 2, 3, 4 and 5 hours. Minimum protein content in cell wall was obtained at 4 and 5 hours extraction and the residue protein was 17.54% (w/w). The protein was further extracted by using 0.3% (w/v) Savinase^â 16L TYPE EX for 4 hours or 0.5% (w/v) Alcalase^â 2.4L for 4 hours which resulted in cell wall with protein contain of 6.18 and 8.81% (w/v) and β-glucan content of 82.44 and 76.88% (w/w), respectively. Removal of lipid in cell wall was carried out by using the mixture of hexane and methanol and pure methanol under reflux; the defatted yeast cell wall contained 0.15% (w/w) and  β-glucan as a water holding capacities, oil holding capacities and emulsifying stabilizer were studied compared with commercial products, it was found the brewer’s yeasts β-glucan from this study had the same water holding capacities, oil holding capacities but emulsion stabilizing capacity was comparable to the commercial product.