Glucoamylase (GA) from Aspergillus flavus HBF34 strain was partially purified 120 folds using starch affinity chromatography. Two isoenzymes (GA1 and GA2) were identified by polyacrylamide gel electrophoresis (PAGE) zymography. Sodium dodecyl sulfate (SDS)-PAGE analysis revealed that one of the enzymes consist of one subunit and the other, two subunits. The optimum pH of the purified GA was 6.0 and the optimum temperature was 60 C. GA was found to be stable at temperatures up to 50 C and at a pH range between 3.0 and 9.0. Km and Vmax values of the enzymes were determined using soluble potato starch, glycogen, amylopectin and amylose as substrates and calculated to be 0.046,