Determination of the transgalactosylation activity of Aspergillus oryzae beta-galactosidase: effect of pH, temperature, and galactose and glucose concentrations

作者：Vera Carlos; Guerrero Cecilia; Illanes Andres^*

来源：Carbohydrate Research, 2011, 346(6): 745-752.

DOI：10.1016/j.carres.2011.01.030

• beta-Galactosidase
• Transgalatosylation
• Galacto-oligosaccharides
• Lactulose
• Inhibition

The catalytic potential of beta-galactosidase is usually determined by its hydrolytic activity over natural or synthetic substrates. However, this method poorly predicts enzyme behavior when transglycosylation instead of hydrolysis is being performed. A system for determining the transgalactosylation activity of beta-galactosidase from Aspergillus oryzae was developed, and its activity was determined under conditions for the synthesis of galacto-oligosaccharides and lactulose. Transgalactosylation activity increased with temperature up to 55 degrees C while the effect of pH was mild in the range from pH 2.5 to 5.5, decreasing at higher values. The effect of glucose and galactose on transgalactosylation activity was also assessed both in the reactions for the synthesis of galacto-oligosaccharides and lactulose and also in the reaction of hydrolysis of o-nitrophenyl beta-D-galactopiranoside. Galactose was a competitive inhibitor and its effect was stronger in the reactions of transgalactosylation than in the reaction of hydrolysis. Glucose was a mild activator of beta-galactosidase in the reaction of hydrolysis, but its mechanism of action was more complex in the reactions of transgalactosylation, having this positive effect only at low concentrations while acting as an inhibitor at high concentrations. This information is relevant to properly assess the effect of monosaccharides during the reactions of the synthesis of lactose-derived oligosaccharides, such as galacto-oligosaccharides and lactulose.

• 出版日期2011-5-1