Kefir Research Articles

Analysis of Kefir Grains from Different Regions of the Planet Using High-Throughput Sequencing.

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.

Production of kurut (kurt) using probiotic Lacticaseibacillus rhamnosus GG strain in combination with a yeast isolated from Kazakhstan kurut

Kuruts are traditional acid-coagulated fermented dairy products in semisolid or dried form. The present study used probiotic Lacticaseibacillus rhamnosus GG (LGG) and Cryptococcus laurentii yeast, a kurut isolate, to make mixed-fermentation kurut. In addition, kurut was fermented with L. rhamnosus GG as starter. Kurut was also fermented with kefir grains representing a traditional way to initiate the fermentation. The viability of probiotic L. rhamnosus GG strain and physicochemical properties of semisolid kurut products were monitored during storage over 22 days at 4°C. The tested probiotic strain showed viability higher than 7 log CFU/mL in both mixed-fermentation and single-strain-fermentation kurut during storage at 4°C. When prepared with probiotic bacteria, the syneresis values of mixed-fermentation kurut were lower (p > 0.05) than single-strain-fermentation kurut. The flavor and overall acceptability scores for kurut prepared using the combination of probiotic LGG and C. laurentii yeast as starter culture were higher than for the other samples at 11 days of storage (p > 0.05). Taking the above into consideration, probiotic kuruts obtained by milk fermentation inoculated with co-culture of L. rhamnosus GG and C. laurentii yeast could be potential probiotic products to be produced in the dairy industry.

Microbiomes of Kefir Grains From Regions of Historical Origin and Their Probiotic Potential

Kefir has attracted interest in the scientific community due to its beneficial properties, such as: improving digestion, antimicrobial action, hypocholesterolemic, antihypertensive, and anti-inflammatory effects, control of plasma glucose levels, antioxidant, anticarcinogenic, and antiallergic activity. The comparative characteristics of the microbiomes of kefir grains (KG) obtained from the regions of their historical origin (Caucasus and China) is given. The diversity of their compositions was shown using classical microbiological and molecular genetic methods, including high-throughput sequencing of V4-region of bacterial 16S rRNA genes and the ITS1 region of the 18S-ITS1-5.8S-ITS2-28S complex of yeast rRNA. The species of the Lactobacillus genus are the most common of the bacterial cultures in kefir grains, while the most common of the yeast cultures is Pichia fermentans , as well as the rarer species of Yarrowia lipolytica and Galactomyces candidus .

Biological activities and physiochemical properties of low-fat and high-fat coconut-based kefir

Coconut milk is a rich source of several nutrients and free of lactose which can be an excellent alternative to cow milk for lactose-intolerant consumers. This study aimed to investigate the potential of producing coconut-based kefir beverages using low-fat and high-fat coconut milk. The beverage was evaluated for pH values, titratable acidity, peptide content, antibacterial activity, antioxidant activity, and rheological properties. The results showed a significant increase in lactic acid bacteria and yeast after the fermentation for 24 h. The pH of the low-fat and high-fat coconut milk kefir declined to 3.77 ± 0.01 and 3.84 ± 0.01, respectively. The titratable acidity increased to 0.75 ± 0.04% and 0.66 ± 0.01% lactic acid, respectively. The fermentation with kefir grains significantly increased the peptides content of low-fat coconut milk to 0.7 ± 0.016 mg mL−1, and for high-fat coconut milk, it increased to 0.571 ± 0.038 mg mL−1. In addition, low-fat coconut kefir showed strong antibacterial growth inhibition and antioxidant activity. The apparent viscosity of the kefir beverage decreased with increasing shear rate and was higher at a lower temperature. The low-fat coconut kefir's flow behavior index (n) and consistency index (K) were 0.216 and 0.223. The consistency index (K) was 3.067 and 0.576 Pa s n at 5 and 25 °C, respectively. Moreover, the (n) values of high-fat coconut kefir were recorded as 0.422 and 0.581, and (K) values as 0.33 and 1.245 Pa s n. Therefore, low-fat coconut milk can be an alternative to cow milk in producing kefir-based beverages.

The effects of fermentation with water kefir grains on two varieties of tigernut (Cyperus esculentus L.) milk

This study investigated the effect on the brown and yellow varieties of tigernut milk after fermentation with water kefir grain, and then determined the properties and bioactive components of the resulting tigernut kefirs. The yellow variety of tigernut milk had higher glucose, fructose and sucrose concentrations. The lactococci and lactobacilli counts for the tigernut kefirs reached from 8.71 to 8.78 log CFU/ml and from 6.94 to 8.14 log CFU/ml, respectively. The presence of yeast in tigernut kefir is related to water kefir grain microbiota. Tigernut kefirs contained from 2.44 to 2.96 g/L of lactic acid. The stepwise discriminant analysis demonstrated that succinic acid, folate, vitamin E, and iron contents were found to be discriminating attributes of the tigernut milk varieties. In contrast, benzoic acid, quercetin and vitamin K were discriminating attributes for tigernut kefirs. The total phenolic content of the samples varied from 77.82 to 101.77 mg of gallic acid equivalents (GAE) L−1; the brown variety of tigernut kefir had a lower total phenolic content than the yellow variety. An increase in the total antioxidant capacity was noticed after kefir fermentation. The extensive findings of this study demonstrated that tigernut milk from both varieties provide an excellent environment for the growth of water kefir microbiota.

The metagenomic composition of water kefir microbiota

Water kefir is a sourish, slightly alcoholic, and carbonated drink in which different dried fruits are added to the sucrose-containing solution and fermented using water kefir grains. Kefir grains in polysaccharide structure include Lactic Acid Bacteria (LAB), yeasts, and/or acetic acid bacteria that use sucrose as an energy source. Water kefir grains contain a varying number of microbiota with various microbiological compositions, depending on their origin. The present study metagenomically determined the microbial flora of water kefir grains obtained from five different commercial sources. According to the metagenomic composition results obtained with the selected specific primers, the dominant bacterial species in water kefir grains were determined to be Lactobacillus ruminis and Bacillus methanolicus, and the most common species were Lactococcus lactis and Enterococcus faecium. Achromobacterx ylosoxidans were detected in only one sample. Pichia kudriavzevii was the dominant yeast species, and Saccharomyces cerevisiae, Eremothecium cymbalariae, Candida glabrata, and Ogataea parapolymorph were seen as common yeast species in all kefir yeasts. Other yeast types showed differences. It was revealed that natural water kefir grains contained probiotic bacteria including Lactobacillus ruminis, so kefir produced using these types of grains can exert functional properties. Another important conclusion is that, albeit, at low levels, kefir grains contain strains that have probiotic potential, including Lactococcus lactis, Enterococcus faecium, and Lactobacillus reuteri.

Microbial diversity associated with Tibetan kefir grains and its protective effects against ethanol-induced oxidative stress in HepG2 cells

Tibetan kefir grains (TKGs) and their products are multi-functional health supplements. This study aimed to explore the diversity of microorganisms in Tibetan kefir grains (TKGs) and the protective effect of isolates on ethanol-treated HepG2 cells. Metagenomic sequencing showed that Lactobacillus Kefiranofaciens was the most abundant bacterial taxon, while Kluyveromyces marxianus was the most abundant fungal taxum in TKGs. Thirteen isolates belonging to the genera Lactobacillus (3), Fructobacillus (1), Lactococcus (1), Pediococcus (1), Acetobacter (3), and Kluyveromyces (4), were obtained by cultivation methods. KEGG pathway analysis revealed that K. marxianus contributed four KEGG pathways related to hepatic disease. LC-MS analysis revealed CFEs are rich sources of peptides amino acids, lipids and amino acids. Cell-free extracts (CFEs) prepared from K. marxianus protected HepG2 cells from ethanol-induced injury. Treatment with CFEs from isolates YG-4 and YK-6 significantly increased cell viability and reduced ethanol cytotoxicity. Also, YG-4 and YK-6 decreased reactive oxygen species, malondialdehyde and alanine aminotransferase, increased superoxide dismutase and catalase, and reduced apoptosis and loss of potential in mitochondria. In addition, CFEs, especially YG-4-derived CFEs, decreased ethanol-induced inflammatory cytokines (IL-6, IL-17 and TNF-α), while YG-4 and YK-6 CFEs upregulated the regulatory cytokine IL-4. Thus, YG-4 had the most significant anti-inflammatory effects by reducing ethanol-induced oxidative stress. These isolates’ properties may provide new avenues to use K. maxianus in alcoholic liver disease.

Sustainable, carbonated, non-alcoholic beverages using leftover bread

Bread is one of the most discarded foods in the world. In this study, we aimed to create sustainable, carbonated, non-alcoholic drinks by using leftover bread. For that purpose, we used leftover bread (non-treated, and pre-treated with Aspergillus oryzae) from the Michelin 3-star Azurmendi restaurant. The drinks were obtained by fermentation, via three different microorganisms (Saccharomyces bayanus, kombucha symbiotic culture of bacteria and yeast “SCOBY”, and water kefir grains). We investigated (i) the optimal fermentation processes, (ii) the physicochemical properties of the obtained drinks, and (iii) consumer liking by a sensory study. The drink elaborated by S. bayanus was preferred by consumers in the liking test.

Characterization and Immunological Activity of Exopolysaccharide from Lacticaseibacillus paracasei GL1 Isolated from Tibetan Kefir Grains

Two exopolysaccharide fractions (GL1-E1 and GL1-E2) of Lacticaseibacillus paracasei GL1 were isolated with the molecular weights of 3.9 × 105 Da and 8.2 × 105 Da, respectively. Both fractions possessed mannose, glucose, and galactose in molar ratios of 1.16:1.00:0.1, and 3.81:1.00:0.12, respectively. A structural arrangement of two fractions was proposed by methylation, one-dimensional and two-dimensional nuclear magnetic resonance experiments. The backbone of GL1-E1 consisted of →4)-α-D-Glcp(1→, →3,4)-α-D-Manp(1→, →3,6)-α-D-Manp(1→, →6)-α-D-Manp(1→, and →6)-α-D-Galp(1→ with α-D-Glcp at branching point. The backbone of GL1-E2 consisted of →4)-α-D-Glcp(1→, →3,4)-α-D-Manp(1→, →3,6)-α-D-Manp(1→, →6)-α-D-Manp(1→, →6)-α-D-Galp(1→, and →4)-β-D-Manp(1→, and the side chain also consisted of α-D-Manp residue. In addition, the differential scanning calorimetry (DSC) analysis indicated that both GL1-E1 and GL1-E2 had good thermal stability. Furthermore, the two fractions could promote the viability of RAW264.7 cells and exert an immunomodulatory role by enhancing phagocytosis, increasing nitric oxide (NO) release and promoting the expression of cytokines.

Yeast cell wall polysaccharides in Tibetan kefir grains are key substances promoting the formation of bacterial biofilm

This study investigated the interaction among Kluyveromyces marxianus G-Y4 (G-Y4), Lacticaseibacillus paracasei GL1 (GL1) and Lactobacillus helveticus SNA12 (SNA12) that isolated from Tibetan kefir grains. Additionally, the effects of G-Y4 on the growth and biofilm formation of GL1 and SNA12 were determined. The results indicated that G-Y4 promoted the growth of GL1 and SNA12 and improved their biofilm-forming ability. Furthermore, the dead cells of G-Y4 were found that could enhance bacterial biofilm formation, and the cell wall polysaccharide (CWPS) produced by G-Y4 was performed to be key substances that promote the formation of bacterial biofilms. Moreover, the structure of soluble cell wall polysaccharides (SCWP) and insoluble cell wall polysaccharide (NCWP) of G-Y4 were studied to determine their contribution to biofilm formation. Results showed that G-Y4-SCWP was α-mannan with the main chain of a →6)-α-d-Manp-(1→ unit and the branch structure of →2)-α-d-Manp-(1. At the same time, G-Y4-NCWP was a glucan rich in β-(1→3), β-(1→2), or β-(1→4) linkages.

The effect of kefir fermentation on the protein profile and the monoterpenic bioactive compounds in goat milk

Changes in proteins and monoterpenic bioactive components as a result of fermenting milk of different goat breeds with kefir grains were investigated. SDS-PAGE was performed to determine the protein profile. Percentages of αS-casein (αS-CN), β-CN, κ-CN, β-lactoglobulin, bovine serum albumin and other peptides were measured. Hair goat milk contained 5.7% αS1-CN, whereas cow milk contained 37% αS1-CN and the kefir fermentation process decreased the αS1-CN content in goat milk. Milk samples from extensively fed goats contained higher β-CN. Proteolysis increased due to the kefir fermentation, and new peptides were formed. Monoterpenic compounds such as carvacrol and thymol were present (0.14 mg 100 g−1 and 0.17 mg 100 g−1, respectively) in the Hair goat milk. After kefir fermentation, carvacrol was present (0.30 mg 100 g−1) in Hair goat kefir. This study is the first to show significant differences in proteolysis and monoterpenic bioactive components in the goat milk and kefir.

Ameliorative Effect of Saccharomyces cerevisiae JKSP39 on Fusobacterium nucleatum and Dextran Sulfate Sodium-Induced Colitis Mouse Model

The aim of this study was to evaluate the ability of the Saccharomyces cerevisiae strain with probiotic properties isolated from Tibetan kefir grains to ameliorate Fusobacterium nucleatum (Fn) infection and dextran sulfate sodium (DSS) treatment-induced murine model of colitis. The tolerance to simulated gastrointestinal conditions, hydrophobicity test, autoaggregation assay, and the antioxidant effect of strains was used to screen one strain with colonization and probiotic potential. The murine model of colitis was established by giving 109 cfu Fn 3 weeks by intragastric administration and 3% DSS in water 1 week before sacrifice. The results indicated that S. cerevisiae JKSP39 (SC) possessed optimal probiotic characteristics in vitro. Supplementation with SC increased the body weight and the expression of anti-inflammatory cytokines (IL-4 and IL-10), while decreasing the disease activity index score and expression of proinflammatory cytokines (TNF-α, IL-6, and IL-17F) in mice undergoing experimental colitis as compared with the colitis model group. Additionally, tight junction proteins and the number of goblet cells per crypt were significantly increased in the SC group, which indicated that the gut barrier was well repaired. The mechanism of SC ameliorating Fn-DSS-induced colitis could be related to the decreased levels of reactive oxygen species (myeloperoxidase, total superoxide dismutase, catalase, H2O2, and malondialdehyde) in the colon, the inhibition of endoplasmic reticulum stress, and the regulation of gut microbiota.

Isolation and screening of high‐quality lactic acid bacteria and yeast strains in kefir grains and preparation of kefir compound fermentation starter

In this study, a total of ten strains of kefir grains were isolated. Lactic acid bacteria and yeast were the research subjects. The final compound fermentation starter was produced using Leu. Mesenteroides, Lactobacillus kefir, Lactobacillus casei, and K. marxianus. The fermented milk of the compound fermentation starter and kefir grains were compared in curding time, acidity, water holding capacity (WHC), ethanol content, the viability of strains, and sensory score. The results showed that the quality of the fermented milk was similar, but the kinds of volatile aroma components in the kefir produced by the compound strain starter were significantly more than those of the kefir grains. The result of this study will help simplify and stabilize the industrial production of kefir, and provide a reference for the development of different flavored yogurt products. Novelty impact statement The dominant kefir grain strains were divided to create the strain compound starter, which can be used in place of kefir grains. It can ferment and produce a milk drink with similar characteristics to traditional kefir grains. The dairy industry can develop using strain starters to overcome the limitations of kefir grains and produce kefir products with consistently high quality.

Evaluation of quality and protein structure of natural water kefir-fermented quinoa protein concentrates

The utilisation of quinoa protein concentrates (QPCs) is limited due to their poor protein digestibility (78.54 %). In this study, QPCs (1 % w/v) were fermented in 5 % (v/v) water kefir grains (WKG) for 5 days at 25 °C. The protein quality of the fermented QPCs was enhanced, whereby the protein digestibility increased significantly (P < 0.05) from 78.54 to 87.67 % after 5 days of fermentation and significant (P < 0.05) changes to the secondary protein structure components (α-helix and random coil). Concurrently, saponins content and pH decreased significantly (P < 0.05). After 2 days of fermentation, the total phenolic content increased significantly (P < 0.05) from 314.5 to 528.8 mg of GAE/100 g. Various phenolic compounds were detected, with epicatechin being the most abundant in the fermented QPCs. WKG fermentation posed as a potential reliable method for enhancing the quality and protein structure of QPCs.

Nanofiltration of the Remaining Whey after Kefir Grains' Cultivation.

Acid whey is derived from fresh cheese. The proteins were isolated by a monolithic ion-exchange column. The remaining whey fraction was used as a starter culture substrate of kefir grains. The aim of this work was, firstly, to study the possibility of column replacement by a UF membrane. If we succeeded, the concentrate would be used as a starter culture substrate of kefir grains. The second part of the research was to purify the remaining solution. The idea was to separate this solution to the permeate and the concentrate by nanofiltration. Further application of both filtration streams was tested as aqueous solutions or dried matter. Chemical and microbiological analyses were performed of both the permeate and the concentrate. The permeate analyses showed that lactose had been fully removed. The aqueous permeate was not stable, mainly due to an increase of total bacteria from 103 to 106 CFU/mL. Therefore, the permeate was spray-dried. The dry permeate was added to the moisture solution in different concentrations. The results showed that up to 0.5% of the dry permeate could be added to the moisturizing solution, with negligible changes in properties having the ability to inhibit acne growth. Anaerobic digestion of industrial sludge was performed with the addition of an aqueous concentrate, which showed improvement in anaerobic fermentation.

The Chemical, Microbiological and Volatile Composition of Kefir-like Beverages Produced from Red Table Grape Juice in Repeated 24-h Fed-Batch Subcultures.

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.

Tailoring the Optimized Fermentation Conditions of SCOBY-Based Membranes and Milk Kefir Grains to Promote Various Functional Properties.

Kombucha culture (named SCOBY-Symbiotic Culture of Bacteria and Yeasts) and milk kefir grains represent multiple consortia of wild microorganisms that include lactic acid bacteria, acetic acid bacteria and yeasts with valuable functional properties. Their fermentative potential provides a wide range of derivate metabiotics (prebiotics, probiotics, postbiotics and paraprobiotics) with valuable in vitro and in vivo benefits. This study targeted the evaluation of the functionality of a co-culture of SCOBY-based membranes and milk kefir grains, used as freeze-dried starter cultures, for the fermentation of a newly formulated medium based on black tea infusion, supplemented with bovine colostrum and sugar, in order to produce bioactive compounds with functional properties. The design and optimization of the biotechnological process were achieved by using the Plackett–Burman experimental design (six factorial points, three center points) and the response surface methodology and central composite design (three factorial points, six axial points and two center points in axial) tools. The statistical analysis and the mathematical modelling of the responses such as the pH, titratable acidity, antioxidant activity and antimicrobial activity (against Bacillus subtilis, Escherichia coli, Staphylococcus aureus and Aspergillus niger) were investigated. Further, the composition of organic acids, polyphenols and flavonoids of the fermented product obtained under the optimized fermentation conditions was also analyzed. The fermentation of the medium containing 6.27% (w/v) bovine colostrum powder, 1.64% (w/v) black tea, 7.5% (w/w) sugar, pH 6.7, with an inoculum based of 0.36% (w/v) milk kefir grains powder and 0.5% (w/v) SCOBY-based membrane (both as freeze-dried culture), at 30 °C, for 5 days, in an aerobic stationary system, revealed an antifungal activity between 80 and 100% against Aspergillus niger, an antibacterial activity of 8–22 mm against Escherichia coli and Bacillus spp. And a titratable acidity of 445 °Th. The chemical composition of the obtained product had a positive impact on the functional properties of the fermented products in terms of the antimicrobial and antioxidant properties.

Acquisition strategies for fermentation processes with a low-cost miniaturized NIR-spectrometer from scratch: Issues and challenges

In recent years, low-cost miniaturized NIR spectrometers have become extremely useful for quantification, discrimination and classification problems in many field and industrial application domains. The dairy industry is showing particular interest in using portable near-infrared spectroscopy as a quality assessment method or process analytical tool but, when coming to at-line, in/on-line possibilities there is no effective and explanatory literature that presents how to approach analytical method development, even though handheld spectroscopy is invaluable to evaluate fermentation as a crucial dairy operation.In this study the potential and limits of a handheld spectrometer (1350–2550 nm) in gaining information about a dairy fermentation process were investigated. Kefir is a fermented dairy product of rising interest, due to its health beneficial effects and the complexity of the process which involved chemicals and physical transformations that bring to the final product starting from milk by inoculating the so-called “kefir grains”; although the chemistry and physics involved are convoluted, the process itself can be easily conducted at home. Handheld spectrophotometers could be helpful to determine early stages of fermentation, before visual changes occur; being a helpful tool for the healthy citizen.Spectral characteristics and variability related to different experimental set-up were explored first, for the starting and ending products and then, during the fermentation. Semi-skimmed milk was used as fermentation media. After adding the kefir grains, spectra were acquired at interval times during the evolution of the process that was carried out under temperature control. To catch all the information included in the spectra and to investigate the possibilities of paving the way for process monitoring, different approaches were employed.

Multi-Objective Optimization of Beverage Based on Lactic Fermentation of Goat’s Milk Whey and Fruit Juice Mixes by Kefir Granules

Numerous fruits are produced in Ecuador, of which about 40% are never eaten. In addition, fresh goat cheeses are in high demand. However, goat cheese generates goat milk whey with high contamination loads, and, therefore, it must be adequately treated before being discharged into ecosystems. This research aims to use a mixture of tree tomato, common strawberry juices, and goat’s milk whey, to be statically fermented by milk and water kefir grains (WKG) for 48 h. For this, a dual mixture design of L-optimal response surface methodology was carried out to find the conditions that maximized all the responses evaluated (lactic-acid bacteria and yeasts concentrations and the overall acceptability assessed on a 7-point scale). Experiments were carried out in San Gabriel, Ecuador. Temperatures during the day and night were 20.2 ± 0.3 °C and 18.7 ± 0.3 °C, respectively. Three conditions were selected, where the highest response values were reached. Complementary experiments demonstrated the validity of the models. When comparing the results of the present study with similar ones carried out previously, higher values were observed in the concentration of yeasts, which seems related to the presence of the WKG. It is concluded that they could be suitable functional beverage candidates.

Adsorption removal of ochratoxin A from milk by Tibetan kefir grains and its mechanism

Tibetan kefir grains (TKG) are microbial symbiotic starters used in various fermented foods. Recent studies have focused on removing toxin contaminants from food using TKG. This work aims to investigate the ability of TKG to remove ochratoxin A (OTA) from milk, the adsorption mechanism of TKG, and the detoxification behavior after exposure to a simulated gastrointestinal tract. The results revealed that TKG and TK removed 90.7% and 92% of OTA, respectively, while the removal capacity of isolates was low and strain-dependent (3.7%–40.7%). The limited OTA removal of freeze-dried TKG (19.6%), TKG kefiran (3.1%), and TKG protein (9.3%) indicated that the matrix network structure was critical in TKG adsorption. The effects of pH, temperature, inoculation percentage, and initial concentration on OTA removal using TKG were investigated. OTA adsorption was consistent with the Freundlich model and the pseudo-second-order kinetic model. In simulated gastric and intestinal fluids, the removal of OTA by TKG reached a maximum at 4 h with 76.1% and 75.6%, respectively. Moreover, only 33.9% of OTA was released from the TKG-OTA complex after a series of washing treatments. These findings support TKG to effectively adsorb OTA contamination in milk while fermenting and can reduce gastrointestinal absorption.