Hidrólise da lactose pela β-galactosidase de Kluyveromyces fragilis: caracterização da enzima

Abstract

The β-galactosidase enzyme from Kluyveromyces fragilis was characterized in the soluble form using lactose 5% w/v found in skimmed powdered milk as substrate. Enzyme diluted 50 times hydrolyzed the lactose in batch reactor of 50 mL capacity. Enzyme activity and its activation energy were determined as a function of temperature and pH. Temperature ranged from 20 to 55oC and pH from 5.5 to 8.0. Activation energy was 9.50 kcal/mol. The energy of deactivation was 33.74 kcal/mol. Although the enzyme presented a high specific activity at 45oC and pH 6.5 (3.312 U/mg protein), values indicate that the best use of the enzymatic activity occur at 40oC or below, with half-life higher than 12 hours. The activation energy increased proportionally to pH increase. Therefore, the activation energy depends on pH and varies according to the origin of the enzyme.