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Entrapping probiotic bacteria in gels with ionic cross-linking is typically achieved with polysaccharides (alginate, pectin, carraghenan). In this study, whey proteins were used for this purpose by carrying out the Ca(2+)-induced gelation of pre-heated whey protein isolate (WPI). A Lactobacillus rhamnosus cell suspension was added in a denatured WPI solution in a 30 : 70 volume ratio. Gelation was carried out by extrusion of the cell suspension in a CaCl(2) solution. Beads of approximately 3 mm diameter were formed. The population in the beads was 8.0 x 10(8) cells g(-1). Entrapment efficiency in gel beads was 96%, with a survival level of 23%. Scanning electron microscopy of beads before freeze-drying showed a tight protein network containing encapsulated Lb. rhamnosus cells homogeneously distributed throughout the matrix. The survival to freeze-drying of the bead-entrapped cells was 41%. Viability of microentrapped cells in a dynamic gastro-intestinal (GI) model was studied and the results were compared to free cells freeze-dried in a milk-based cryoprotective solution, as well as in a pre-denatured WPI solution. Results showed that protein gelation provided protection against acidic conditions in the stomach after 90 min, as well as against bile after 30, 60 and 90 min in the duodenum. Moreover, the milk-based cryoprotective solution was equally effective after 90 min in the duodenum. It is concluded that the gelation of whey proteins induced by Ca(2+) ions can protect the cells against adverse conditions of the GI system. However, certain stages in the entrapment process, particularly extrusion in the solution of CaCl(2), still need to be optimized in order to reduce the mortality of the cells during gelation.

This study investigated the influence of ionic strength, liposome net charge and enzyme concentration on the immobilization of chymotrypsin in liposomes obtained from proliposomes. Depending on ionic strength and chymotrypsin concentration, immobilization efficiencies (IE) as high as 96 and 68% were obtained for liposomes prepared with Pro-lipo 3045 S and Pro-lipo 3080 S respectively. Increasing ionic strength and enzyme concentration resulted in a decrease in IE for both types of liposomes, and this was more pronounced for ionic strength. Relatively high amounts of chymotrypsin were found to be immobilized on the surface of the liposomes. Hydrophobic interactions between chymotrypsin and the hydrophobic tails of the phospholipids during liposome formation were probably responsible for this phenomenon.

The development of a method for the continuous mass production of liposomes is vital for the industrial use of liposomes in food products. The method should be mild enough to prevent denaturation of the encapsulated material, and the materials used for the preparation of the liposomes should be safe and edible. Among the methods available, microfluidization seems to be the most promising. Microfluidization consists of processing emulsions under high pressure through an apparatus called a Microfluidizer. This apparatus also allows the production of another type of lipid vesicle: milkfat-coated microcapsules composed of milkfat and emulsifiers. The main advantages of microfluidization include the continuous production of large quantities of lipid vesicles without dissolving the phospholipids in organic solvents. These vesicles could be used in various food products for a variety of objectives. The addition of encapsulated material in liposomes or in milkfat-coated microcapsules to cheese milk resulted in a retention of 80-90% of the vesicles in the cheese, compared to only 2-4% if the material was added directly to the milk in the unencapsulated form. Liposomes and milkfat-coated microcapsules could be used as enzyme carriers to accelerate cheese ripening, or as carriers for flavouring systems to improve the organoleptic properties of low-fat cheeses or to impart distinctive flavours to new speciality cheeses. These microcapsules could also be used in food products to avoid undesirable side-reactions during food processing, or to supplement food products with nutritious additives.