Two forms of glucoamylase (GI and GII) from starch-grown Lipomyces kononenkoae CBS 5608 mutant were purified to apparent homogeneity by means of ultrafiltration, Sephacryl S-200 gel filtration and DEAF Sephadex A-50 chromatography. The apparent molecular weight was calculated as ca. 150 kDa for GI and ca. 128 kDa for GII, respectively. Both enzymes were glycoproteins with isoelecMc points of 5.6 (GI) and 5.4 (GII). They had a pH optimun of 4.5 and were stable from pH 5 to 8. The temperature optimum for both enzymes was 6090, but they were rapidly inactivated above 70?. The Km values toward starch were estimated to be 6.57 ㎎ per ㎖ for GI and 4.52 ㎎ per ㎖ for GII, and the V_(max) values were 16.28 μM per ㎎ for GI and 32.25 μM per ㎎ for GII, respectively. The K_m and Vmax values of GII for a- or p-cyclodextrin were estimated to be 0.15 ㎎ per ㎖ and 2.0 ㎎ per ㎖, respectively (K_m) and 1.02 μM per ㎎ or 1.02 μM per ㎎, respectively (V_(max)). Neither enzyme exhibited pullulanase activity but they released only glucose from starch or cyclodextrin. Amino acid analysis indicated that both glucoamylases were enriched in proline and acid amino acids. Glucoamylase GII strongly cross-reacted with a monoclonal antibody rnised against GI enzymes, and the two enzymes shared very similar amino acid composition. Western blot analysis indicated that L. kononenkoae CBS 5608 mutant produced two forms of glucoamylase on starch, and that synthesis of them was subject to glucose repression.