| Summary: | The application of β-galactosidase in the fermentation of milk enables the acquirement of lower levels of lactose that are tolerated by lactose maldigesters and can reduce the nutritional consequences of avoiding dairy products. The present study evaluated the viability of the fortification of lactose-free prebiotic Greek yogurt formulas with whey protein concentrate (WPC). Two rotational central composite designs (RCCDs) were applied: one to perform the hydrolysis of the whey protein concentrate and another for the yogurt formulations (α = 2 with 2 central points and 4 axial points). Two β-galactosidase enzymes obtained from <i>Kluyveromyces lactis</i> were used. The content of lactose, glucose, galactose, and lactic acid were determined in the WPC, milk (pasteurized and powdered), and yogurts. The three best formulations regarding the attributes’ viscosity, syneresis, firmness, and elasticity were sensorially evaluated by using a nine-point hedonic scale. A microbiological analysis was performed after 48 h of yogurt production. The characterization of the products and the comparison of the results obtained were evaluated using the Student’s <i>T</i> test and the analysis of variance with Tukey’s test (<i>p</i>-values < 0.05). The application of a lactose-free WPC promoted viscosity, firmness, and elasticity. The syneresis was reduced, and whey increased the protein and calcium content. Lactose-free WPC can be used as a partial substitute for skimmed powdered milk in yogurts. The obtained results are encouraging with respect to the production of lactose-free Greek yogurts by the dairy industry.
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