Effect of prebiotics on thermally acclimatized lactobacilli cultures and their application as synbiotics in RTD fruit drinks.

In this study, the effect of prebiotics such as fructooligosaccharides (FOS), galactooligosaccharides (GOS), isomaltooligosaccharides (IMO), and inulin on the probiotic biomass and its probiotic properties were studied for thermally acclimatized Lactobacillus helveticus (H-45) and Lacticaseibacillus casei N (N-45) strains at 45 â„ƒ using adaptive laboratory evolution method. Among the prebiotics studied, GOS was found to be more suitable for synbiotic preparation. The tolerance of lactobacilli cultures H-45 and N-45 in the presence of acid and bile were 4.79 and 8.60% and 2.84 and 4.65% higher than their wild-type strains (H-37 and N-37). Similarly, H-45 and N-45 showed an increase in survivability of 5.29 and 8.63% under simulated gastric conditions and 9.21 and 7.70% under simulated intestinal conditions than H-37 and N-37. Propionic acid yield increased by 0.65-fold in N-45 compared to N-37 in the presence of GOS as a prebiotic, whereas H-37 showed 0.26-fold higher propionic acid production than H-45. Thermally acclimatized strain N-45 showed better survivability under stress conditions than H-45. The synbiotic combination of N45 + GOS was spray-dried using corn starch (CS) as carrier material to obtain spray-dried synbiotic powder (N45 + CS + GOS). This synbiotic powder was added to the ready-to-drink (RTD) fruit drinks prepared from five fruit-flavoured squashes (pineapple, orange, grape, mango, and lemon ginger). The varied amounts of added synbiotic powder did not significantly alter the physicochemical properties of the fruit drinks. Hence, synbiotic formulation N45 + GOS + CS may find application in developing various functional foods.

Development of Thermotolerant Lactobacilli Cultures with Improved Probiotic Properties Using Adaptive Laboratory Evolution Method.

Probiotics play a significant role in functional foods. Heat stress and dehydration are the two principal mechanisms leading to inactivation and loss of probiotics viability in its production. There is a need to develop an industrial organism to withstand higher temperatures during its processing and storage. This current study aims to develop thermotolerant strains of Lacticaseibacillus casei N (N) and Lactobacillus helveticus NRRL B-4526 (H) by acclimatizing the wild-type strains to the higher temperature of 45 °C by adaptive laboratory evolution. A two-fold increase in biomass was observed in both acclimatized strains up to the 200th generation, which subsequently remained stable after 500 generations. The morphological change of these acclimatized strains was observed under scanning electron microscopy. Also, there was an increase in probiotic attributes of these acclimatized strains compared to their wild-types. Among two acclimatized strains, L. casei N-45 had shown higher tolerance in the acidic pH 3.0 (89.31%), the bile of 0.3% (84.45%), simulated gastric juice (79.12%), and simulated intestinal juice (73.86%). There was also an increase in salt tolerance (NaCl), radical scavenging activity, autoaggregation, coaggregation, and hydrophobicity of these adapted strains. The total protein profiling using 2D gel electrophoresis reveals the differences in protein expressions between wild-type and acclimatized strains. Specific protein spots from acclimatized strains of H-45 and N-45 were further subjected to MALDI-TOF MS/MS. Some of the identified proteins were recognized to play a role in RNA chaperones and protein synthesis during stress conditions.