Purification and characterization of an extracellular glucoamylase from the yeast Candida tsukubaensis CBS 6389.
The starch-degrading yeast Candida tsukubaensis CBS 6389 secreted amylase at high activity when grown in a medium containing soluble starch. The extracellular alpha-amylase activity was very low. The major amylase component was purified by DEAE-Sephadex A-50 chromatography and Ultrogel AcA 44 gel filtration and characterized as a glucoamylase. The enzyme proved to be a glycoprotein with a molecular weight of 56 000. The glucoamylase had a temperature optimum at 55 degrees C and displayed highest activity in a pH range of 2.4-4.8. Acarbose strongly inhibited the purified glucoamylase. Debranching activity was present as demonstrated by the release of glucose from pullulan.
D002175:Candida / Q000201:enzymology*; D005087:Glucan 1,4-alpha-Glucosidase / Q000302:isolation & purification* / Q000378:metabolism; D005959:Glucosidases / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D013816:Thermodynamics
D005959:Glucosidases; D005087:Glucan 1,4-alpha-Glucosidase