Development and Evaluation of Probiotic Delivery Systems using the Rennet-induced Gelation of Milk Proteins

The aims of this study were to develop a milk protein-based probiotic delivery system using a modified rennet-induced gelation method and to determine how the milk protein concentration level and pH, which can affect the rennet-induced intra- and inter-molecular association of milk proteins, affect the physicochemical properties of the probiotic delivery systems, such as the particle size, size distribution, encapsulation efficiency, and viability of probiotics in simulated gastrointestinal tract. To prepare a milk protein-based delivery system, skim milk powder was used as a source of milk proteins with various concentration levels from 3% to 10% (w/w) and rennet was added to skim milk solutions followed by adjustment of pH from 5.4 or 6.2. <italic>L</italic>.<italic> rhamnosus</italic> GG was used as a probiotic culture. In confocal laser scanning microscopic images, globular particles with a size ranging from 10 μm to 20 μm were observed, indicating that milk protein-based probiotic delivery systems were successfully created. When the milk protein concentration was increased from 3% to 10% (w/w), the size of the delivery system was significantly (p <italic>&lt; </italic>0.05) increased from 27.5 μm to 44.4 μm, while a significant (<italic>p </italic>&lt; 0.05) increase in size from 26.3 μm to 34.5 μm was observed as the pH was increased from 5.4 to 6.4. An increase in the milk protein concentration level and a decrease in pH led to a significant (<italic>p</italic> &lt; 0.05) increase in the encapsulation efficiency of probiotics. The viability of probiotics in a simulated stomach condition was increased when probiotics were encapsulated in milk protein-based delivery systems. An increase in the milk protein concentration and a decrease in pH resulted in an increase in the viability of probiotics in simulated stomach conditions. It was concluded that the milk protein concentration level and pH were the key manufacturing variables affecting the physicochemical properties of milk protein-based probiotic delivery systems.