Abstract
A new method for the separative determination of filamentous fungi amylase was described. The procedure was as follows : 5ml of 1.0% soluble starch solution, 3 ml of M/10-acetate buffer solution (pH 5.0) and 1 ml of enzyme solution in 50 ml Elrenmyer flask, was incubated at 37°C for 30 minutes. After addition of 1 ml of 1 N-NaOH solution to stop the reaction, dextrose formed in the reaction mixture was determined by the Tauber-Kleiner's method (glucoamylase activity). By using the Fehling-Lehmann-Schoolr's method in stead of the Tauber-Kleiner's method, free reducing sugar in the reaction mixture was determined (total amylase activity). Glucoamylase activity subtracted from total amylase activity was defined α-amylase activity. One unit of amylase activity was defined as the amount of enzyme which produces 100 μM of glucose from starch per one hour at 37°C.
