Kefir milk, known for its high nutritional value and health benefits, is traditionally produced by fermenting milk with kefir grains. These grains are a complex symbiotic community of lactic acid bacteria, acetic acid bacteria, yeasts, and other microorganisms. However, the intricate coexistence mechanisms within these microbial colonies remain a mystery, posing challenges in predicting their biological and functional traits. This uncertainty often leads to variability in kefir milk\'s quality and safety. This review delves into the unique structural characteristics of kefir grains, particularly their distinctive hollow structure. We propose hypotheses on their formation, which appears to be influenced by the aggregation behaviors of the community members and their alliances. In kefir milk, a systematic colonization process is driven by metabolite release, orchestrating the spatiotemporal rearrangement of ecological niches. We place special emphasis on the dynamic spatiotemporal changes within the kefir microbial community. Spatially, we observe variations in species morphology and distribution across different locations within the grain structure. Temporally, the review highlights the succession patterns of the microbial community, shedding light on their evolving interactions.Furthermore, we explore the ecological mechanisms underpinning the formation of a stable community composition. The interplay of cooperative and competitive species within these microorganisms ensures a dynamic balance, contributing to the community\'s richness and stability. In kefir community, competitive species foster diversity and stability, whereas cooperative species bolster mutualistic symbiosis. By deepening our understanding of the behaviors of these complex microbial communities, we can pave the way for future advancements in the development and diversification of starter cultures for food fermentation processes.

Probiotics are known for their health-promoting resources and are considered as beneficial microorganisms. The current study focuses on the isolation, and on a complete in vitro and in vivo characterization, of yeast and lactic acid bacteria acquired from traditional homemade kefir in order to assess their potentiality as probiotic candidates. In particular, the isolates Pichia kudriavzevii Y1, Lactococcus lactis subsp. hordniae LAB1 and Lactococcus lactis subsp. lactis LAB2 were subjected to in vitro characterization to evaluate their suitability as probiotics. Resistance to acid and bile salts, auto-aggregation, co-aggregation, hydrophobicity, and biofilm production capability were examined, as well as their antioxidant activity. A safety assessment was also conducted to confirm the non-pathogenic nature of the isolates, with hemolysis assay and antibiotic resistance assessment. Moreover, mortality in the invertebrate model Galleria mellonella was evaluated. Current findings showed that P. kudriavzevii exhibited estimable probiotic properties, placing them as promising candidates for functional foods. Both lactic acid bacteria isolated in this work could be classified as potential probiotics with advantageous traits, including antimicrobial activity against enteric pathogens and good adhesion ability on intestinal cells. This study revealed that homemade kefir could be a beneficial origin of different probiotic microorganisms that may enhance health and wellness.

The utilization of pea proteins (PPs) is limited due to their relatively low protein digestibility (∼81%) compared to animal-based proteins, such as whey. The present investigation involved the fermentation of PPs at a concentration of 1% (w/v) using 5% (w/v) water kefir for 60 h at 25°C to improve the functional properties of PPs. The results showed a significant (p < 0.05) increase in lactic acid and acetic acid production during fermentation. These findings suggest that PPs can be effectively fermented using water kefir as a starter culture for the increased protein digestibility of PPs. The PP conformation underwent modifications, including secondary and tertiary protein structure alterations. The total phenolic compounds increased throughout the fermentation, reaching around 695.32 ± 15 mg gallic acid equivalent/100 g after 24 h of fermentation. Furthermore, the fermentation process has culminated in significant (p < 0.05) changes in the surface charge and hydrophobic properties of the fermented PPs, from -38.1 to -45.73 and 362.7 to 550.2, respectively. Fermentation using water kefir is a promising technique for improving the digestibility, protein structure, and nutritional values of PPs.

Soy protein isolate (SPI) has become a promising plant-based material as an animal protein products alternative. However, its application was limited due to the weak gelling properties. To investigate the effect of kefir fermentation on SPI gels properties, SPI-polysaccharide gels was produced by unfermented and kefir-fermented SPI using different concentration of KGM, chitosan, and calcium chloride in this study. Characterization of fermented SPI gels showed that fermentation by kefir grains can be applied to improve the textural strength, mechanical structure, and thermal characteristics of SPI gels. Compared to unfermented SPI gels, the water-holding capacity was remarkably enhanced to 63.11% and 65.71% in fermented SPI-chitosan gels. Moreover, the hardness of fermented SPI-KGM gels were significantly increased to 13.43 g and 27.11 g. And the cohesiveness and resilience of fermented-KGM gels were also improved than unfermented samples. Results of rheological characterization and thermogravimetric analysis revealed the strengthened mechanical features and higher thermal stability of fermented SPI gels. Additionally, the main role of hydrophobic interactions and secondary structure variations of SPI gels were demonstrated by intermolecular force measurements, Fourier-transform infrared spectroscopy, and X-ray diffraction. Moreover, the network structure was observed more compact and homogeneous performed by microstructural images in fermented SPI gels. Therefore, this research provided a novel approach combining multi-species fermentation with protein gelation to prepare SPI gel materials with improved nutrition and structural properties.

Soybeans possess unexpected flavors and are difficult to be absorbed by the gastrointestinal tract. Kefir grain fermentation provides diverse strains and bioactive compounds, which may enhance flavor and bioaccessibility. Third-generation sequencing was applied to analyze the microbial diversity in milk and soybean kefir grains in this study. In both types of kefir grains, the most common bacterial genus was Lactobacillus, and their fungal communities were dominated by Kazachstania. Lactobacillus kefiranofaciens was the most abundant species in kefir grains, while Lactobacillus kefiri showed a higher proportion in soybean kefir grains. In addition, the quantification of free amino acids and volatile flavor compounds in soybean solution and soybean kefir have shown the increased content of glutamic acid and a decreased amount of unpleasant beany flavor compounds, demonstrating that the nutritive value and sensory properties of soybean can be improved by kefir grain fermentation. Finally, the bioconversion of isoflavones during fermentation and in vitro digestion was evaluated, suggesting that fermentation is beneficial for aglycone formation and absorption. To conclude, kefir fermentation is proposed to change the microbial structure of kefir grains, promote the nutritional value of soybean-based fermented products, and provide possible solutions for the development of soybean products.

The taxonomic composition and spatial localization of yeast and bacteria in kefir grains (KG) obtained for study from different regions of the planet were investigated. The diversity of their microbiome has been demonstrated by high-throughput sequencing of bacterial 16S rRNA genes and the ITS1 region of the 18S-ITS1-5.8S-ITS2-28S complex of yeast rRNA. It has been established that the main representatives of the complex community of KG from different regions are lactic acid bacteria (LAB; lactobacilli, lactococci, and Leuconostoc spp. in different ratios) and different types of yeast of the genus Kazachstania (family Saccharomycetaceae). Acetic acid bacteria and a small percentage of yeast Kluyveromyces marxianus were detected in the KG from Tibet, and yeast Pichia kluyveri was detected in the KG from Ossetia.

Lacticaseibacillus paracasei (formerly Lactobacillus paracasei) is a nomadic lactic acid bacterium (LAB) that inhabits a wide variety of ecological niches, from fermented foodstuffs to host-associated microenvironments. Many of the isolated L. paracasei strains have been used as single-strain probiotics or as part of a symbiotic consortium within formulations. The present study contributes to the exploration of different strains of L. paracasei derived from non-conventional isolation sources-the South African traditional fermented drink mahewu (strains MA2 and MA3) and kefir grains (strains KF1 and ABK). The performed microbiological, biochemical and genomic comparative analyses of the studied strains demonstrated correlation between properties of the strains and their isolation source, which suggests the presence of at least partial strain adaptation to the isolation environments. Additionally, for the studied strains, antagonistic activities against common pathogens and against each other were observed, and the ability to release bioactive peptides with antioxidant and angiotensin I-converting enzyme inhibitory (ACE-I) properties during milk fermentation was investigated. The obtained results may be useful for a deeper understanding of the nomadic lifestyle of L. paracasei and for the development of new starter cultures and probiotic preparations based on this LAB in the future.

The aim of this work was to study the production of kefir-like beverages via the fed-batch fermentation of red table grape juice at initial pHs of 3.99 (fermentation A) and 5.99 (fermentation B) with kefir grains during 4 repeated 24-h fed-batch subcultures. All kefir-like beverages (KLB) were characterized by low alcoholic grade (≤3.6%, v/v) and lactic and acetic acid concentrations. The beverages obtained from fermentation B had lower concentrations of sugars and higher microbial counts than the KLB obtained in fermentation A. Additionally, the KLB samples from fermentation B were the most aromatic and had the highest contents of alcohols, esters, aldehydes and organic acids, in contrast with the nonfermented juice and KLB from fermentation A. These results indicate the possibility of obtaining red table grape KLB with their own distinctive aromatic characteristics and high content in probiotic viable cells, contributing to the valorization of this fruit.

The analysis of the literature on the microbiome composition and metabolic properties of kefir available at the RSCI and Web of Science was carried out. Kefir has been used by humans for centuries. It is a useful product of mixed lactic and alcoholic fermentation, produced using evolutionally established associative cultures, collected in an aggregated state termed kefir grains. General characterization of kefir grains from the territorial zones of different continents (Russia, Europe, Asia, and America) is provided. The methods for differentiation and identification of individual species are described, as well as their interactions within the community. The diversity of microbial composition of kefir grains depending on local cultivation conditions and storage processes is shown. The microorganisms present in kefir have a number of properties that determine their metabolism, interaction in the community, beneficial effects on human health and immune system, which is important for the prevention and control of bacterial and viral infections, especially during the COVID-19 pandemic.

Proteins not only serve as a nitrogen source for microorganisms but are the main skeleton of kefir grains. After subculturing in goat milk for 4 months, proteins and peptides in three kefir grains from China, Germany, and the United States were analyzed. Except for the S-layer protein from special Lactobacillus sp., αs1-casein, αs2-casein, and β-casein from goat milk were found in kefir grains. These proteins could form aggregates through a covalent interaction with polysaccharides to maintain the morphological stability of the grains. Furthermore, they were highly related to the microbiota in kefir grains. Additionally, a number of hydrophilic/hydrophobic peptides that were hydrolyzed by extracellular proteases were found from kefir grains. A correlation may exist between peptides and Lactobacillus sp. in kefir grains. Bioactive peptides, including DKIHPF, LGPVRGPFP, and QEPVLGPVRGPFP, were found from these kefir grains. The results indicated that goat milk as a substrate affects the protein and peptide composition of kefir grains.