Evaluation of lactose oxidase as enzymatic antifungal control for Penicillium spoilage in yogurt

Abstract

In this study, we investigated the antifungal activity of lactose oxidase (LO) as a potential biopreservative in dairy products. Our study objectives were to screen antifungal activity of LO against common mold strains, to detect the minimum inhibitory level of LO against the same strains, and to understand how LO affects the pH and lactic acid bacteria (LAB) counts in set yogurt. Five mold strains (Penicillium chrysogenum, Penicillium citrinum, Penicillium commune, Penicillium decumbens, and Penicillium roqueforti) were used throughout study. These strains were previously isolated from dairy manufacturing plants. Throughout the study, yogurts were stored at 21 ± 2°C for 14 d. Antifungal activity of LO was screened using 2 enzyme levels (1.2 and 12 g/L LO) against selected strains on the surface of a miniature laboratory set-yogurt model. For all tested strains, no visible mold growth was detected on the surface of yogurts covered with LO compared with control yogurt without LO. The minimum inhibitory level of LO against each strain was further investigated using 4 enzyme levels (0.12, 0.48, 0.84, and 1.2 g/L LO) on the miniature laboratory set-yogurt model. We detected 0.84 g/L LO as the minimum level inhibiting visible hyphal growth across strains. The minimum inhibitory level of LO varied for each individual strain. To study the effect of LO on the pH of yogurt, miniature laboratory set-yogurt models were covered with different enzyme levels (0.12, 0.48, 0.84, 1.2, and 12 g/L LO). At d 14, a difference was detected comparing pH values of treatments to control with no LO. Commercial lowfat set yogurt was used to study the effect of LO on LAB survival when yogurt surface was covered with 0.84 g/L LO under the same experimental conditions. Control with no LO was included. At d 14, 3 levels of catalase were added (0, 0.01, and 0.1%) to each treatment. To enumerate LAB, homogenized samples were plated on de Man, Rogosa, and Sharpe agar and incubated. Yogurts with 0.84 g/L LO had lower LAB counts compared with control yogurts, and catalase level did not have a significant effect on LAB counts. Our results demonstrated potential antifungal efficacy of LO against common spoilage organisms in dairy products with residual lactose and relatively low pH. Manufacturers should establish efficacy of LO against mold strains of interest and determine the effects of LO on organoleptic properties and LAB survival in set yogurt.