Study of the probiotic properties of Lacticaseibacillus casei subsp. casei NCIM 5752 and the optimization of whey-based media for the production of its biomass using response surface methodology.

In this study, Lacticaseibacillus casei NCIM 5752, a new isolate has been explored for probiotic properties and has shown significant bile salt hydrolase activity and cholesterol-reducing activity (56.7 ± 0.27%) in the presence of bile salts. It also tested negative for the production of lecithinase and gelatinase, indicating its non-pathogenic nature. The test strain was able to tolerate pH of 2.0 and 3.0 with 63.42 and 94.7% of the cells survived after 3 h. L. casei showed auto-aggregation of 85.3% and surface hydrophobicity of 22.5% in xylene and 19.4% in hexane. Paneer whey was explored as a basic raw material for alternative media formulation for growing lactic acid bacteria. Paneer whey was found to contain lactose (4.15%), protein (0.42%), and rich in mineral content. Response surface methodology was employed to optimize the medium composition with three independent variables yeast extract (X1), dextrose (X2), and dipotassium hydrogen phosphate (X3), and the response-Y was set to biomass obtained in terms of log CFU/ml. They were supplemented to paneer whey medium for growing this strain. The second-order polynomial regression model predicted that the maximum cell mass production of 11.30 ± 0.5 log CFU/ml at optimal composition of 16.22 g/L of yeast extract, 19.31 g/L of dextrose, and 2.12 g/L of dipotassium hydrogen phosphate in paneer whey medium. Experiments were conducted to validate the RSM results, and the biomass achieved was 11.27 ± 0.50 log CFU/ml, which is in close agreement with the yield predicted by the RSM. By applying the fermentation strategy, the biomass was increased to 5.56 ± 0.34 g/L dry cell weight corresponding to 11.58 ± 0.24 log CFU/ml. The newly optimized media was significantly cost-effective and produced 26.45% more biomass than the conventional MRS media. This optimized media may find application for the large-scale biomass production of probiotics. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03899-z.

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.

Solid-state fermentation of Jatropha seed cake for optimization of lipase, protease and detoxification of anti-nutrients in Jatropha seed cake using Aspergillus versicolor CJS-98.

This study focused on the solid-state fermentation of Jatropha seed cake (JSC), a byproduct generated after biodiesel production. Presence of anti-nutritional compounds and toxins restricts its application in livestock feed. The disposal of the JSC is a major environmental problem in the future, due to the generation of huge quantity of JSC after biodiesel extraction. Hence the JSC was assessed for its suitability as substrate for production and optimization of lipase and protease from Aspergillus versicolor CJS-98 by solid-state fermentation (SSF). The present study was also focused on the biodetoxification of anti-nutrients and toxins in JSC. The SSF parameters were optimized for maximum production of lipase and protease. Under the optimized conditions, the JSC supplemented with maltose and peptone (2%), adjusted to pH 7.0, moisture content 40%, inoculated with 1 × 10(7) spores per 5 g cake and incubated at 25°C, produced maximum lipase, 1288 U/g and protease, 3366 U/g at 96 h. The anti-nutrients like phytic acid (6.08%), tannins (0.37%), trypsin inhibitors (697.5 TIU/g), cyanogenic glucosides (692.5 µg/100 g), and lectins (0.309 mg/ml), were reduced to 1.70%, 0.23%, 12.5 TIU/g, 560.6 µg/100 g and 0.034 mg/ml respectively. The main toxic compound phorbol esters content in the JSC was reduced from 0.083% to 0.015% after SSF. Our results indicate that viability of SSF to utilize the huge amount of seed cake generated after extraction of biodiesel, for production of industrial enzymes and biodetoxification of anti-nutrients, toxins.