João Miguel Rocha

A variety of foods fermented with lactic acid bacteria (LAB) serve as dietary staples in many countries. The incorporation of health-promoting probiotics into fermented milk products can have profound effects on human health. Considering the health benefits of Yakult, the current study was undertaken to develop an enriched Yakult-like fermented skimmed milk drink by the addition of two probiotic strains, namely Lacticaseibacillus casei (Lc) and Lacticaseibacillus rhamnosus (Lr). The prepared drinks were compared in terms of various parameters, including their physicochemical properties, proximate chemical composition, mineral estimation, microbial viable count, antioxidant activity, and sensory evaluation. Each strain was employed at five different concentrations, including 1% (T1), 1.5% (T2), 2% (T3), 2.5% (T4), and 3% (T5). The prepared Yakult samples were stored at 4 °C and analyzed on days 0, 7, 14, 21, and 28 to evaluate biochemical changes. The findings revealed that the concentration of the starter culture had a significant (p ≤ 0.05) impact on the pH value and moisture and protein contents, but had no marked impact on the fat or ash content of the developed product. With the Lc strain, Yakult’s moisture content ranged from 84.25 ± 0.09 to 85.65 ± 0.13%, whereas with the Lr strain, it was from 84.24 ± 0.08 to 88.75 ± 0.13%. Protein levels reached their highest values with T5 (3% concentration). The acidity of all treatments increased significantly due to fermentation and, subsequently, pH showed a downward trend (p ≤ 0.05). The total soluble solids (TSS) content decreased during storage with Lc as compared to Lr, but the presence of carbohydrates had no appreciable impact. The drink with Lc exhibited a more uniform texture and smaller pore size than Yakult with Lr. Except for the iron values, which showed an increasing trend, the contents of other minerals decreased in increasing order of the added probiotic concentration used: 1% (T1), 1.5% (T2), 2% (T3), 2.5% (T4), and 3% (T5). The highest lactobacilli viable count of 8.69 ± 0.43 colony-forming units (CFU)/mL was observed with the T1 Lr-containing drink at the end of the storage period. Regarding the storage stability of the drink, the highest value for DPPH (88.75 ± 0.13%) was found with the T1 Lc drink on day 15, while the highest values for FRAP (4.86 ± 2.80 mmol Fe2+/L), TPC (5.97 ± 0.29 mg GAE/mL), and TFC (3.59 ± 0.17 mg GAE/mL) were found with the T5 Lr drink on day 28 of storage. However, the maximum value for ABTS (3.59 ± 0.17%) was noted with the T5 Lr drink on the first day of storage. The results of this study prove that Lc and Lr can be used in dairy-based fermented products and stored at refrigerated temperatures.

Exopolysaccharides (EPSs) produced by lactic acid bacteria present a particular interest for the food industry since they can be incorporated in foods via in situ production by selected starter cultures or applied as natural additives to improve the quality of various food products. In the present study, 43 strains were isolated from different plant-based fermented foods and identified by molecular methods. The species found were distinctively specific according to the food source. Only six Lactiplantibacillus plantarum strains, all isolated from sauerkraut, showed the ability to produce exopolysaccharide (EPS). The utilization of glucose, fructose and sucrose was explored with regard to EPS and biomass accumulation by the tested strains. Sucrose was clearly the best carbon source for EPS production by most of the strains, yielding up to 211.53 mg/L by strain Lactiplantibacillus plantarum ZE2, while biomass accumulation reached the highest levels in the glucose-based culture medium. Most strains produced similar levels of EPS with glucose and fructose, while fructose was utilized more poorly for biomass production, yielding about 50% of biomass compared to glucose for most strains. Composition analysis of the EPSs produced by strain Lactiplantibacillus plantarum ZE2 from glucose (EPS-1) and fructose (EPS-2) revealed that glucose (80–83 mol%) and protein (41% w/w) predominated in both analyzed EPSs. However, the yield of EPS-1 was twice higher than that of EPS-2, and differences in the levels of all detected sugars were found, which shows that even for the same strain, EPS yield and composition vary depending on the carbon source. These results may be the basis for the development of tailored EPS-producing starter cultures for food fermentations, as well as technologies for the production of EPS for various applications.