Effect of Fluidized Bed Drying, Matrix Constituents and Structure on the Viability of Probiotic Lactobacillus paracasei ATCC 55544 during Storage at 4 °C, 25 °C and 37 °C.

The stabilization of probiotics for application in non-refrigerated food products is a challenging task. In the present study, probiotic Lactobacillus paracasei (Lacticaseibacillus paracasei) ATCC 55544 cells were immobilized in a dairy matrix comprising of whole milk powder, skim milk powder, or milk protein isolate using fluidized bed drying technology. The samples were taken out at different drying stages, with an apparent water activity (aw) of aw 0.5, aw 0.4, and aw 0.3, respectively, and vacuum-packed to maintain the aw and stored at three different temperatures of 4 °C, 25 °C, and 37 °C. The study evaluated the impact of matrix constituents, milk fat, protein, and carbohydrate on the viability of encapsulated probiotic L . paracasei ATCC 55544 during storage for 1 month. The whole milk powder matrix provided superior protection to the bacteria. Confocal Laser Scanning Microscopy (CLSM) was used to investigate the structure of the immobilizing matrix and the location of the probiotic L. paracasei cells embedded within the matrix. The CLSM study revealed that the probiotic bacterial cells are mostly embedded as clusters beneath the top layer. We hypothesize that the biofilm-like structure, together with the protective whole milk powder matrix, helps to retain the superior viability of probiotic cells during storage at non-refrigerated storage conditions of 25 °C and 37 °C.

Importance of intact secondary protein structures of cell envelopes and glass transition temperature of the stabilization matrix on the storage stability of probiotics.

Lactobacillus reuteri LR6 cells were stabilized using a novel combination of wet granulation and fluidized-bed-drying techniques. The stabilized cells were stored at 37 °C and at two water activity (aw) levels (0.11 & 0.30). Superior storage stability was recorded in the lower aw environment, supported by a stronger glassy matrix when skim milk powder was used as the excipient. The initial viable cell populations of the samples stabilized in different matrices ranged from 8.3 to 9.1 log CFU/g. At the end of the storage period, the viable cell populations were reduced to 6.7 to 7.3 log CFU/g at aw 0.11 and to 6.1 to 6.6 CFU/g when the aw was maintained at 0.30. Fourier transform infrared spectroscopic examination of the cell envelopes revealed substantial dissimilarities between samples at the beginning and at the end of the storage period, which indicated alteration in the secondary protein structures of the cell envelope and also correlated well with the loss in cell viability. In milk-powder-based matrices, adjusting the aw to 0.30 resulted in a weaker or no glassy state whereas the same matrices had a high glass transition temperature at aw 0.11. This strong glassy matrix and low aw combination was found to enhance the bacterial stability at the storage temperature of 37 °C. Scanning electron microscopy revealed the formation of corrugated surfaces and blister-type deformations on the cell envelopes during the stabilization process.