O objetivo deste trabalho foi estudar a influência de parâmetros reacionais na obtenção via enzimática de galacto-oligossacarídeos (GOS), utilizando-se a enzima comercial β-galactosidase de Kluyveromyces lactis (Lactozym® 3000 L), tendo como substrato a lactose. Foi proposto um planejamento experimental 2³, verificando a influência da temperatura (30 a 40 °C), da concentração de lactose (200 a 400 mg.mL-1) e da concentração de enzima (5 a 10 U.mL-1) no desempenho da reação enzimática. Os ensaios foram conduzidos a 180 rpm e pH 7,0 (tampão fosfato de sódio 0,1 M). A enzima Lactozym® 3000 L apresentou atividade de transgalactosilação, tendo sido atingido rendimento do processo igual a 41,9% e concentração de GOS de 167,5 mg.mL-1 no sistema reacional composto por 400 mg.mL-1 de lactose e 5 U.mL-1 de enzima a 30 °C em 14 h de reação. Nessa condição, a conversão de lactose foi de 65,0%. Maior concentração de lactose foi favorável ao mecanismo de transgalactosilação, enquanto que, em menores concentrações, o mecanismo hidrolítico predominou.

The main goal of this work was to study the influence of the reaction parameters on the enzymatic production of galactooligosaccharides (GOS) using the commercial β-galactosidase from Kluyveromyces lactis (Lactozym® 3000 L) and lactose as the substrate. A 2³ experimental design was proposed, verifying the influence of the temperature (30 to 40 °C), lactose concentration (200 to 400 mg.mL-1) and enzyme concentration (5 to 10 U.mL-1) on the performance of the enzymatic reaction. The assays were carried out at 180 rpm and pH 7.0 (0.1 M sodium phosphate buffer). The enzyme Lactozym® 3000 L presented transgalactosylation activity, reaching a yield of 41.9% and GOS concentration of 167.5 mg.mL-1 in a reaction system composed of 400 mg.mL-1 of lactose and 5 U.mL-1 of enzyme at 30 °C and 14 h of reaction. Under these conditions the lactose conversion was 65.0%. Higher lactose concentrations favored the transgalactosylation mechanism, whereas the hydrolytic mechanism dominated with lower concentrations.

The optimization of a traditional technique of cellular disruption by abrasion was carried out and a process using ultrasonic waves associated with glass pearls to extract beta-galactosidase from Kluyveromyces marxianus proposed. In the first case, the effects of the diameter and weight of the pearls in relation to the volume of cellular suspension and amount of time for cellular disruption were evaluated. The efficiency of the new process of cellular disruption was evaluated by varying the length of time of sonification and comparing with the method of abrasion under the same conditions. The proposed method can be efficiently applied to obtain beta-galactosidase at laboratory scale.