Production of intracellular gamma-glucosidase in brewery waste by: saccharomyces carlsbergensis

RESUMO

Saccharomyces carlsbergensis strain ATCC 9080 was grown on acid hidrolyzed brewery waste. The sustrate contains 22 mg/ml of total carbohydrates of which approximately 60% represents reducing sugars such as maltose and glucose. Optimal conditions for substrate hydrolysis were 1.5% w/v sulfuric acid, 20 minutes and temperature of 110 grados C. Maximum enzyme production was obtained in a medium containing 0.05% w/v yeast extract at 35 grado C and at an initial pH of 6. Addition of nitrogen sources was unnecessary. Optimal conditions for gamma-glucosidase extraction from yeast included pH 7.5, 30 grados C, and 10 minutes of sonication. Maximum hydrolysis of PNPG occured at pH 7.5, temperature between 30 grados C and 25 minutes of incubation. Michaelis-Menten constants and Vmax were 1 mM, 0.56 mol/min/mg of protein on P-nitrophenly-gamma-D-glucophyranoside and 5 mV, 5 mol/min/mg of protein on maltose respectively