Improvement in Epigenetic Aging Clock Induced by BioBran Containing Rice Kefiran in Relation to Various Biomarkers: A Pilot Study.

Many lifestyle-related diseases such as cancer, dementia, myocardial infarction, and stroke are known to be caused by aging, and the WHO's ICD-11 (International Classification of Diseases, 11th edition) created the code "aging-related" in 2022. In other words, aging is irreversible but aging-related diseases are reversible, so taking measures to treat them is important for health longevity and preventing other diseases. Therefore, in this study, we used BioBran containing rice kefiran as an approach to improve aging. Rice kefiran has been reported to improve the intestinal microflora, regulate the intestines, and have anti-aging effects. BioBran has also been reported to have antioxidant effects and improve liver function, and human studies have shown that it affects the diversity of the intestinal microbiota. Quantitative measures of aging that correlate with disease risk are now available through the epigenetic clock test, which examines the entire gene sequence and determines biological age based on the methylation level. Horvath's Clock is the best known of many epigenetic clock tests and was published by Steve Horvath in 2013. In this study, we examine the effect of using Horvath's Clock to improve aging and report on the results, which show a certain effect.

Invited review: Milk kefir microbiota-Direct and indirect antimicrobial effects.

Kefir is a fermented dairy product with well recognized probiotic properties. Recently, consumer interest in fermented products with probiotic microorganisms has increased due to the accumulating evidence of the effects of kefir microorganisms on the modulation of gut microbiota and their antimicrobial activity. Although the health properties of kefir have been reviewed in other works, the present review addresses the antimicrobial effects of kefir microbiota and associated compounds. The antimicrobial activity of kefir microorganisms could derive from different mechanisms. The microorganisms' capacity to adhere to the intestinal epithelium, preventing the adhesion of pathogens, and their immunomodulation properties are among the mechanisms suggested. Bacteria and yeast isolated from kefir have been shown to have in vivo and in vitro antimicrobial activity against enteropathogenic bacteria and spoilage fungi. However, most reports have focused their approach on single-strain antimicrobial properties; evaluation of antimicrobial activity of cocultures of kefir microbiota and their potential mechanisms of action has been neglected. Kefir microbiota and associated compounds have shown promising antimicrobial effects; however, more research needs to be done to discern the mechanisms of action.

Magnetic Biocatalysts of Pectinase: Synthesis by Macromolecular Cross-Linker for Application in Apple Juice Clarification.

RESEARCH BACKGROUND: Pectinase enzyme has become a valuable compound in beverage industry. One of the most significant concepts to overcome the drawbacks of using industrial enzymes is their immobilization. In the present study, magnetic chitosan microparticles were utilized as a substrate for pectinase immobilization. New methods of enzyme immobilization involve the use of non-chemical cross-linkers between the enzyme and the substrate. The aim of this study is to immobilize the pectinase enzyme using polyaldehyde kefiran as a macromolecular cross-linker on magnetic particles. EXPERIMENTAL APPROACH: Pectinase was immobilized in four steps: relative oxidation of kefiran and its application as a cross-linker, production of magnetic iron(II) iron(III) oxide (Fe3O4) microparticles, coating of magnetic Fe3O4 microparticles with chitosan, and immobilization of the enzyme on the substrate, prepared by the use of oxidized kefiran cross-linker. Parameters such as cross-linking concentration, time and ratio of chitosan magnetic microparticles to enzyme were optimized. Fourier-transform infrared spectroscopy (FTIR), dynamic light scattering, transmission electron microscopy, and vibrating sample magnetometer were used to identify the groups and investigate the structures. The biochemical properties (stability of enzyme activity at different pH, temperature and time), enzyme reusability, kinetic parameters (K m and ν max) and apple juice turbidity, using free and immobilized pectinase enzymes, were also measured. RESULTS AND CONCLUSIONS: Cross-linker concentration, cross-linking time and the ratio of magnetic Fe3O4 microparticles with chitosan to enzyme were important factors in activity recovery of pectinase. FTIR analysis correctly identified functional groups in the structures. The results showed that after enzyme stabilization, the particle size and molecular mass, respectively, increased and decreased the magnetic saturation strength. According to the thermal kinetic study, the activity of the immobilized pectinase was higher than of its free form. The findings of this study indicate excellent stability and durability of the immobilized pectinase. Finally, a magnetic pectinase micro-biocatalyst was used to clarify apple juice, which reduced turbidity during processing. NOVELTY AND SCIENTIFIC CONTRIBUTION: This study investigates the usage of kefiran oxidized as a new cross-linker for the immobilization of pectinase enzyme. Magnetic pectinase micro-biocatalyst has a good potential for industrial applications in the food industry, with high thermal stability.

Fermented product of rice with Lactobacillus kefiranofaciens induces anti-aging effects and heat stress tolerance in nematodes via DAF-16.

Rice kefiran is superior in functionality, has high concentration of mucilaginous polysaccharide, and low lipid content, compared to conventional kefiran. However, reports on its physiological functionality, especially studies on life expectancy and aging, in model organisms are rare. In this study, nematodes were used as model organisms that were fed rice kefiran, along with Escherichia coli OP50, as a result of which, the lifespan of nematodes was extended and age-related retardation of mobility was suppressed. It also increased the heat stress resistance in nematodes. Experiments using daf-16 deletion mutant revealed that rice kefiran functions via DAF-16. Thus, this study revealed the longevity, anti-aging and heat stress tolerance effects of rice kefiran in nematodes.

Co-production of functional exopolysaccharides and lactic acid by Lactobacillus kefiranofaciens originated from fermented milk, kefir.

Kefiran is a functional exopolysaccharide produced by Lactobacillus kefiranofaciens originated from kefir, traditional fermented milk in the Caucasian Mountains, Russia. Kefiran is attractive as thickeners, stabilizers, emulsifiers, gelling agents and also has antimicrobial and antitumor activity. However, the production costs of kefiran are still high mainly due to high cost of carbon and nitrogen sources. This study aimed to produce kefiran and its co-product, lactic acid, from low-cost industrial byproducts. Among the sources tested, whey lactose (at 2% sugar concentration) and spent yeast cells hydrolysate (at 6 g-nitrogen/L) gave the highest kefiran of 480 ± 21 mg/L along with lactic acid of 20.1 ± 0.2 g/L. The combination of these two sources and initial pH were optimized through Response Surface Methodology. With the optimized medium, L. kefiranofaciens produced more kefiran and lactic acid up to 635 ± 7 mg/L and 32.9 ± 0.7 g/L, respectively. When the pH was controlled to alleviate the inhibition from acidic pH, L. kefiranofaciens could consume all sugars and produced kefiran and lactic acid up to 1693 ± 29 mg/L and 87.49 ± 0.23 g/L, respectively. Moreover, the fed-batch fermentation with intermittent adding of whey lactose improved kefiran and lactic acid productions up to 2514 ± 93 mg/L and 135 ± 1.75 g/L, respectively. These results indicate the promising approach to economically produce kefiran and lactic acid from low-cost nutrient sources.

Rheology and microstructure of kefiran and whey protein mixed gels.

The effect of kefiran on cold-set gelation of whey protein isolate (WPI) at 25 °C was studied using rheological measurements and environmental scanning electron microscopy (ESEM). The gelation of samples was induced by the addition of glucono-δ-lactone to the dispersions. WPI concentration was maintained at 8% (w/v) and the concentration of kefiran varied from 0 to 0.08% (w/v). According to rheological measurements, the addition of kefiran into WPI dispersions resulted in a significant increase in the gel strength, the yield stress, and the shear stress values at the flowing point. The gelling point and gelation pH of samples decreased significantly with an increase in kefiran concentration. ESEM micrographs showed that the presence of kefiran played an important role in the microstructure formation of gels. The microstructure of kefiran-WPI mixed gels was more compact and dense, compared to the WPI gel. Depletion interactions between kefiran and whey protein aggregates can be regarded as the chief factor which was responsible for these effects. The present work demonstrated that rheological and microstructural properties of acid-induced whey protein gels were improved by the addition of kefiran.

Gelling ability of kefiran in the presence of sucrose and fructose and physicochemical characterization of the resulting cryogels.

In this work, the influence of sucrose and fructose on the gel-forming capacity of kefiran was investigated as well as the physicochemical characteristics of the resulting gels. The addition of sugar to gel-forming solutions did not alter the pseudoplastic flow properties of kefiran solutions and after one freeze-thaw cycle translucent gels with high water-holding capability were obtained. A highly porous matrix was revealed by microscopy whose pore size varied with sugar concentration. Sucrose and fructose had different effects on the rheological characteristics of sugar-kefiran gels. An increment in the strength of the gels with progressive concentrations of sucrose was evidenced by an increase in the elastic modulus (G'), indicating that sucrose reinforces the binding interactions between the polymer molecules (p ≤ 0.05). A drastic reduction in elastic modulus occurred, however, when 50.0 % w/w sucrose was added to kefiran gels, resulting in less elasticity. In contrast, when fructose was added to kefiran gels, elastic modulus decreased slightly with progressive sugar concentrations up to 10 %, thereafter increasing up to 50 % (p ≤ 0.05). Supplementation with up to 30 % sugar contributed to water retention and increased the viscous modulus. The relative increment in the elastic and viscous moduli elevated the loss tangent (tanδ) depending on the type and concentration of sugar. Sugars (sucrose, fructose) present in the matrix of the polysaccharide networks modified water-polymer and polymer-polymer interactions and consequently changed the gels' physicochemical characteristics, thus allowing the possibility of selecting the appropriate formulation through tailor-made kefiran cryogels.

Characterization of the new biodegradable WPI/clay nanocomposite films based on kefiran exopolysaccharide.

Physico-mechanical, thermal and structural characteristics of nanocomposite film composed of kefiran-whey protein isolate (WPI)-montmorillonite (MMT; 1, 3 and 5 % w/w) were studied. Incorporation of MMT significantly affected the mechanical attributes of the kefiran-WPI films. The tensile strength and Young's modulus increased and the percentage of elongation at break decreased as the MMT content increased. Moisture content, moisture absorption and water solubility decreased as the MMT concentration increased. Differential scanning calorimetry indicated that the glass transition temperature for kefiran-WPI film was -12.5 °C and was noticeably affected by an increase in MMT. X-ray diffraction analysis showed formation of an exfoliated structure with the addition of small amounts of MMT to the kefiran-WPI matrix. Intercalation and some exfoliation occurred up to 5 % (wt) increase in MMT. Scanning electron microscopy demonstrated ideal dispersion for MMT nanoparticles into the structure of the bio-nanocomposite films.

Development and characterization of kefiran-gelatin bio-nanocomposites containing Zhumeria majdae essential oil nanoemulsion to use as active food packaging in sponge cakes.

Active packaging films based on kefiran-gelatin were developed and characterized using Zhumeria majdae essential oil nanoemulsion (ZMEO-NE) at concentrations of 0 (control), 1, 2 and 3 %. The main compounds of the essential oil (EO) of Zhumeria majdae (ZM) plant were linalool (61.44 %) and camphor (20.67 %). Adding the ZMEO-NE to the films decreased permeability to water vapor (from 7.82 × 10-7 to 4.09 × 10-7 g·m/m2·Pa·h), ultimate tensile strength (from 38.44 to 33.48 MPa), percentage of light transmission, and increased thickness (from 0.085 to 0.121 mm), opacity (from 2.11 to 2.79), and elongation at break (from 19.97 to 34.73 %), and changed color parameters. The establishment of hydrogen bonds between the ZMEO-NE and polymer network was confirmed. The ZMEO-NE decreased the storage modulus and glass transition temperature. Distinct variations in the films' surface morphology and a reduction in the crystalline structure were observed due to the presence of the ZMEO-NE. Elevating the concentration of the ZMEO-NE increased antioxidant capabilities of the films. The films incorporating the ZMEO-NE exhibited notable antibacterial efficacy against Staphylococcus aureus (reductions ≥4 log CFU/cm2) and Escherichia coli (reductions ≥2 log CFU/cm2) bacteria. The films also demonstrated suitable antifungal properties during storage of sponge cakes for 16 days.

Application of culture-dependent and culture-independent methods for the identification of Lactobacillus kefiranofaciens in microbial consortia present in kefir grains.

The biological and technological characteristics of kefiran as well as its importance in grain integrity led us to analyze the microbial kefir grain consortium with focus on Lactobacillus kefiranofaciens. The presence of L. kefiranofaciens in the nine kefir grains studied was demonstrated by denaturing gradient gel electrophoresis. By culture dependent methods applying a methodology focused on the search of this species, 22 isolates with typical morphology were obtained and identified applying a combination of SDS-PAGE of whole cell proteins, (GTG)5-PCR and sequence analysis of the housekeeping gene encoding the α-subunit of bacterial phenylalanyl-tRNA synthase (pheS). This polyphasic approach allowed the reliable identification of 11 L. kefiranofaciens, 5 Lactobacillus paracasei, 4 Lactobacillus kefiri and 2 Lactobacillus parakefiri isolates. Isolated L. kefiranofaciens strains produced polysaccharide in strain-dependent concentrations and EPS produced by them also differed in the degree of polymerization. The isolation and accurate identification of L. kefiranofaciens is relevant taking into account the important role of this microorganism in the grain ecosystem as well as its potential application as starter in food fermentations.

Effect of caseinate salt addition on the structural characteristics of kefiran systems.

Sodium caseinates-kefiran systems were studied to explore whether any potential interactions between them might exist. The study was performed using low-deformation rheological techniques, which were dynamic and creep tests. The systems were prepared under various experimental conditions such as heating and acidification. Besides, the structure development of the systems in relation to time was also monitored using oscillatory shear rheometry. The results indicated that the structural characteristics of the systems were mainly affected by the state of the caseinates such as the formation of aggregates and to a lesser degree by the interactions of kefiran molecules with the caseinates. Freeze-thaw treatment produced cryogels with good thermal stability and fairly satisfactory mechanical properties. The morphology of the caseinate-kefiran systems was also investigated by means of confocal laser scanning microscopy.

Development of Conjugated Kefiran-Chondroitin Sulphate Cryogels with Enhanced Properties for Biomedical Applications.

Hydrogels based on natural polysaccharides can have unique properties and be tailored for several applications, which may be mainly limited by the fragile structure and weak mechanical properties of this type of system. We successfully prepared cryogels made of newly synthesized kefiran exopolysaccharide-chondroitin sulfate (CS) conjugate via carbodiimide-mediated coupling to overcome these drawbacks. The freeze-thawing procedure of cryogel preparation followed by lyophilization is a promising route to fabricate polymer-based scaffolds with countless and valuable biomedical applications. The novel graft macromolecular compound (kefiran-CS conjugate) was characterized through 1H-NMR and FTIR spectroscopy-which confirmed the structure of the conjugate, differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA)-which mirrored good thermal stability (degradation temperature of about 215 °C) and, finally, gel permeation chromatography-size exclusion chromatography (GPC-SEC)-which proved an increased molecular weight due to chemical coupling of kefiran with CS. At the same time, the corresponding cryogels physically crosslinked after the freeze-thawing procedure were investigated by scanning electron microscopy (SEM), Micro-CT, and dynamic rheology. The results revealed a prevalent contribution of elastic/storage component to the viscoelastic behavior of cryogels in swollen state, a micromorphology with micrometer-sized open pores fully interconnected, and high porosity (ca. 90%) observed for freeze-dried cryogels. Furthermore, the metabolic activity and proliferation of human adipose stem cells (hASCs), when cultured onto the developed kefiran-CS cryogel, was maintained at a satisfactory level over 72 h. Based on the results obtained, it can be inferred that the newly freeze-dried kefiran-CS cryogels possess a host of unique properties that render them highly suitable for use in tissue engineering, regenerative medicine, drug delivery, and other biomedical applications where robust mechanical properties and biocompatibility are crucial.

Influence of Three Extraction Methods on the Physicochemical Properties of Kefirans Isolated from Three Types of Animal Milk.

Kefiran is a heteropolysaccharide biopolymer usually extracted from kefir grains cultured in cow milk. Due to the lack of information on exopolysaccharides from other types of animal milk, in the present study, cow, buffalo and goat milks were used as raw materials for fermentation. The kefiran extractions from kefir grains were carried out with cold water (method I), hot water (method II) and mild heated water-ultrasound (method III), and then the recovery yield and the physicochemical properties of the kefirans were evaluated to establish the influence of both the extraction conditions and the type of milk. The highest yield was recorded for the cow kefiran using method III (4.79%). The recoveries of goat and buffalo kefirans with methods II and III were similar (2.75-2.81%). Method I had the lowest yields (0.15-0.48%). The physicochemical characteristics were studied with Fourier Transform-Infrared Spectroscopy (FT-IR), Scanning Electron Microscopy (SEM), and Differential Scanning Calorimetry (DSC). Fourier-transform infrared spectroscopy showed the same qualitative profile for all the samples, regardless of the method and the type of milk, confirming that the extraction methods did not affect the chemical structure of the kefirans. Otherwise, the thermal and morphological features of the samples showed differences according to both the type of the milk and the extraction method. The kefiran samples were very thermally stable, having a temperature of degradation (Td) in the range from 264 to 354 °C. The resulting morphological and thermal differences could lead to different practical applications of kefirans in the fields of nutrition and pharmacology.

Impact of Kefiran Exopolysaccharide Extraction on Its Applicability for Tissue Engineering and Regenerative Medicine.

Kefiran is an exopolysaccharide produced by the microflora of kefir grains used to produce the fermented milk beverage kefir. The health-promoting and physicochemical properties of kefiran led to its exploration for a range of applications, mainly in the food industry and biomedical fields. Aiming to explore its potential for tissue engineering and regenerative medicine (TERM) applications, the kefiran biopolymer obtained through three different extraction methodologies was fully characterized and compared. High-quality kefiran polysaccharides were recovered with suitable yield through different extraction protocols. The methods consisted of heating the kefir grains prior to recovering kefiran by centrifugation and differed mainly in the precipitation steps included before lyophilization. Then, kefiran scaffolds were successfully produced from each extract by cryogelation and freeze-drying. In all extracts, it was possible to identify the molecular structure of the kefiran polysaccharide through 1H-NMR and FTIR spectra. The kefiran from extraction 1 showed the highest molecular weight (~3000 kDa) and the best rheological properties, showing a pseudoplastic behavior; its scaffold presented the highest value of porosity (93.2% ± 2), and wall thickness (85.8 µm ± 16.3). All extracts showed thermal stability, good injectability and desirable viscoelastic properties; the developed scaffolds demonstrated mechanical stability, elastic behavior, and pore size comprised between 98-94 µm. Additionally, all kefiran products proved to be non-cytotoxic over L929 cells. The interesting structural, physicochemical, and biological properties showed by the kefiran extracts and cryogels revealed their biomedical potential and suitability for TERM applications.

Chromatographic profile, in silico and in vivo study of the pharmacokinetic and toxicological properties of major constituent present in kefir, the kefiran.

Kefiran is a polysaccharide present in kefir grains that have been widely explored due to its potential health benefits. The objective of this work was to characterize and quantify the components present in the ethanolic extract of milk kefir grains; to study its pharmacokinetic and toxicological properties in silico and evaluate the acute toxicity of the kefiran in zebrafish. The prediction of pharmacokinetic properties was performed by QikProp software. In silico toxicity assessment was performed using the DEREK (deductive estimate of risk from existing knowledge) software. In the chromatographic, kefiran was identified as the major component. Results showed that the kefiran had low human oral absorption and intestinal absorption its due poor solubility profile; low logP value, indicating its lipophilicity and the low MDCK and Caco-2 cells permability, and unable to cross the blood-brain barrier. Kefiran did not present any structural warning for in silico toxicity. In zebrafish, the dose of 2,000 mg/kg of kefiran produced nonsignificant alterations in the analyzed organs. It can be said then that kefiran has an acceptable degree of safety for use in the development of drugs or functional foods. Further research such as in vivo testing to confirm its pharmacological potential is currently underway.

Functional Properties of Kefiran in the Medical Field and Food Industry.

Kefir is produced through the fermentation of milk using kefir grain as a starter culture. Kefir grains include heterogeneous microorganisms embedded in a polysaccharide matrix called kefiran which is considered a biofilm; it also has many uses due to its therapeutic values. Kefiran is a microbial exopolysaccharide (EPS) obtained from the flora (acid-lactic bacteria and yeasts) of kefir grains and glucose units, in almost the same proportion. Kefiran has prebiotic nature agitating the growth of probiotics in the gastrointestinal tract of the human entity. It extends certain therapeutic benefits by balancing the microbiota in the intestine. This review presents the most recent advances regarding kefir and kefiran, their cultural condition, biological activities, and potential applications in the health and food industries.

Study of the effect of the addition of plasticizers on the physical properties of biodegradable films based on kefiran for potential application as food packaging.

Spectroscopies analysis indicated that kefiran contains branched hexasaccharide repeating units. Neat kefiran films, 2 and 5% w/w of glycerol, d-glucitol, d-galactitol, d-mannitol, and d-limonene were incorporated as plasticizers. Neat and plasticized kefiran films were characterized by physical, thermal, mechanical, optical, and water solubilization properties. Neat kefiran had a glass transition temperature (Tg) of -20 ± 2 °C and, with the addition of plasticizers between -15 to -17 ± 2 °C. The values were close to the neat kefiran, and the results could be attributed to a lower amount of plasticizer used. The solubility of the glycerol plasticized films increases by 33% and decreased with the concentration of other plasticizers in comparison with the neat kefiran. d-glucitol and d-galactitol decreased the microhardness and Young's Modulus of films around 30% and 74% respectively, obtaining more flexible kefiran films. Kefiran based films could find applications as potential materials in the food-packaging industry.

Synthesis and Characterization of Biocompatible Methacrylated Kefiran Hydrogels: Towards Tissue Engineering Applications.

Hydrogel application feasibility is still limited mainly due to their low mechanical strength and fragile nature. Therefore, several physical and chemical cross-linking modifications are being used to improve their properties. In this research, methacrylated Kefiran was synthesized by reacting Kefiran with methacrylic anhydride (MA). The developed MA-Kefiran was physicochemically characterized, and its biological properties evaluated by different techniques. Chemical modification of MA-Kefiran was confirmed by 1H-NMR and FTIR and GPC-SEC showed an average Mw of 793 kDa (PDI 1.3). The mechanical data obtained revealed MA-Kefiran to be a pseudoplastic fluid with an extrusion force of 11.21 ± 2.87 N. Moreover, MA-Kefiran 3D cryogels were successfully developed and fully characterized. Through micro-CT and SEM, the scaffolds revealed high porosity (85.53 ± 0.15%) and pore size (33.67 ± 3.13 µm), thick pore walls (11.92 ± 0.44 µm) and a homogeneous structure. Finally, MA-Kefiran revealed to be biocompatible by presenting no hemolytic activity and an improved cellular function of L929 cells observed through the AlamarBlue® assay. By incorporating methacrylate groups in the Kefiran polysaccharide chain, a MA-Kefiran product was developed with remarkable physical, mechanical, and biological properties, resulting in a promising hydrogel to be used in tissue engineering and regenerative medicine applications.

An investigation of factors affecting the electrospinning of poly (vinyl alcohol)/kefiran composite nanofibers.

This study aimed to investigate parameters affecting the electrospinning of poly (vinyl alcohol) (PVA)/kefiran composite nanofibers. Accordingly, PVA/kefiran composite nanofibers were produced using the electrospinning of PVA, kefiran blend solutions under various electrospinning parameters (such as applied voltage, nozzle-to-collector distance, and polymer injection rate), and solution parameters (such as the ratio of polymers). PVA and kefiran solutions were prepared in 8% and 6% w/w, respectively. Kefiran was blended with PVA solution in different proportions: 70:30, 60:40, 50:50, 40:60, and 30:70. According to the scanning electron microscope (SEM) images, kefiran mixed with PVA in 40:60 ratios produced the best result in nanofiber production. Then, device parameters such as voltage (12, 15, 18, and 20 kV), distance (120, 150, 170, and 200 mm), and polymer injection rates (1, 1.5, 2, and 2.5 mL/h) were changed. The investigation of SEM images showed that the optimal condition for the fabrication of nanofibers was 18 kV, 200 mm, and 1 mL/h. The nanofibers produced in the optimal condition were uniform without knots or adhesion in a small diameter. It was also found that concentration can be regarded as the most effective parameter affecting the diameter of nanofibers. Moreover, the transmission electron microscopy (TEM) image proved that phase separation did not occur between the two polymers. Graphical abstract: Kefiran biopolymer extracted from fermented milk was used in fabrication of PVA/kefiran composite nanofibers using the electrospinning method.

Octenyl succinylation of kefiran: Preparation, characterization and functional properties.

In this study, kefiran was esterified with octenyl succinic anhydride (OSA). The esterification reaction variables including pH (8.5), kefiran concentration (5% (w/w)), OSA concentration (12% (w/w)), temperature (~38 °C) and reaction time (~80 min) were found as optimum points to achieve the maximum degree of substitution (DS) (0.041 ± 0.002). Kefiran-OSA samples with DS of 0.021 (FDA suggested DS) and 0.041 (maximum DS) were prepared and compared with unmodified kefiran in all experiments. FTIR and 1H NMR spectroscopies proved the grafting of OSA on kefiran structure. XRD analysis revealed that with increase in DS, the physical state of kefiran to be more amorphous. In addition, the esterification modification led to a decrease in the degradation temperature and an increase in the apparent viscosity based on the obtained data from thermal analysis and viscosity measurement. The results of the foaming and emulsifying properties confirmed the improvement in surface properties of the modified kefiran. The frequency sweep test illustrated that with an increase in DS, the viscoelastic behavior of the kefiran cryogels to be more viscous. It can finally be stated that the modification with OSA was a high potential strategy to extend the industrial applications of the kefiran.