Isolation and characterization of exopolysaccharides from kombucha samples of different origins.

Kombucha is prepared by fermenting sugared green or black tea with a symbiotic culture of bacteria and yeast (SCOBY). Some of the bacteria within the SCOBY are known to form exopolysaccharides (EPS) from sucrose. However, it is yet unknown whether water-soluble EPS are formed in kombucha, and if so, which specific EPS are present. Therefore, different kombucha samples were prepared by fermentation of green and black tea with SCOBYs from different manufacturers. Subsequently, the EPS were isolated and characterized by using various chromatographic methods, partial enzymatic hydrolyses and NMR spectroscopy. It was demonstrated that levans with a varying degree of branching at position O1 (4.3-7.9 %) are present, while only trace amounts of glucans were detected. Furthermore, levans isolated from kombucha had a comparably low molecular weight and the content of levan within the kombucha samples varied from 33 to 562 mg levan/L kombucha. Therefore, our study demonstrated that levans are the main EPS type in kombucha and that levan amounts and structures varied when different starter cultures and ingredients were used. Furthermore, we provide a comprehensive data set on the structural variability of levans from kombucha.

Characterization of levan produced by a Paenibacillus sp. isolated from Brazilian crude oil.

A levan-type fructooligosaccharide was produced by a Paenibacillus strain isolated from Brazilian crude oil, the purity of which was 98.5% after precipitation with ethanol and dialysis. Characterization by FTIR, NMR spectroscopy, GC-FID and ESI-MS revealed that it is a mixture of linear ß(2 â†’ 6) fructosyl polymers with average degree of polymerization (DP) of 18 and branching ratio of 20. Morphological structure and physicochemical properties were investigated to assess levan microstructure, degradation temperature and thermomechanical features. Thermal Gravimetric Analysis highlighted degradation temperature of 218 °C, Differential Scanning Calorimetry (DSC) glass transition at 81.47 °C, and Dynamic Mechanical Analysis three frequency-dependent transition peaks. These peaks, corresponding to a first thermomechanical transition event at 86.60 °C related to the DSC endothermic event, a second at 170.9 °C and a third at 185.2 °C, were attributed to different glass transition temperatures of oligo and polyfructans with different DP. Levan showed high morphological versatility and technological potential for the food, nutraceutical, and pharmaceutical industries.

Structures of fructan and galactan from Polygonatum cyrtonema and their utilization by probiotic bacteria.

Polygonatum cyrtonema is a known tonic herb in Chinese Materia Medica, extensively consumed in China, but the structure and activity of its polysaccharide components remain to be clarified. Herein, two new polysaccharides (a fructan and a galactan) were purified from the dried and the processed P. cyrtonema rhizome, respectively. Structural analysis suggested that the fructan consisted of a (2 â†’ 6) linked ß-d-Fruf residues backbone with an internal α-d-Glcp residue and two (2 â†’ 1) linked ß-d-Fruf residues branches, and that the galactan was a (1 â†’ 4)-ß-d-galactan branched with a single ß-d-galactose at C-6 at about every nine residues in its main chain. The bioactive assay showed that the fructan and the galactan remarkably promoted growth of Bifidobacterium and Lactobacillus strains, indicating that they possess prebiotic activity. These findings may help expand the application of the polysaccharides from the tonic herb P. cyrtonema as functional ingredients in food products.

Rheological and emulsifying properties of an exopolysaccharide produced by potential probiotic Leuconostoc citreum-BMS strain.

EPS-BMS, is to our knowledge, the first high molecular weight exopolysaccharide from potential probiotic Leuconostoc citreum-BMS strain that consists on a mixture of α-(1,6)-dextran branched at the third position and ß-(2,6)-levan. This sample exhibited interesting rheological and emulsifying properties under different conditions. Steady shear experiments proved that EPS-BMS had a pseudoplastic behavior without thixotropic properties. Interestingly, pseudoplasticity was maintained even under stress conditions of temperature, pH and salts, which could provide some sensory properties for food products such as mouth feel. Dynamic oscillatory measurements reflected a liquid-like behavior of the sample regardless of the studied EPS concentration, pH, temperature and ionic force. Results related to the emulsifying as well as interfacial properties showed that EPS-BMS had great potential to be applied as emulsifier and/or emulsion stabilizer in both neutral and acidic conditions. Based on the properties reported in this work, EPS-BMS could be potentially applied in the food industry.

Structural elucidation and interfacial properties of a levan isolated from Bacillus mojavensis.

A strain with high exopolysaccharide (EPS) production was isolated from soil and identified as Bacillus mojavensis based on the 16S rRNA gene sequencing and biochemical properties. The EPS produced simultaneously with the growth phase reached a maximum of 22 g/L after attaining a stationary phase with sucrose used as sole carbon source. B. mojavensis EPS (BM-EPS) was recovered, fractionated by ethanol precipitation and analysed by NMR and methylation analyses. The BM-EPS was found to be composed of (ß2 â†’ 6)-Fruf residues, characteristic of a levan, with an average molecular weight of 2.3 MDa. A homogeneous micro-porous and rough structure matrix was observed by SEM of the freeze-dried powdered sample. A concentration-dependent water-soluble nature was observed, with good water (5.3 g/g) and oil (36 g/g) holding capacities. The levan displayed good emulsification activity with excellent stability against food grade oil, thus favoring it as a promising emulsifying agent to food industries.

Characterization of inulin-type fructans from two species of Radix Codonopsis and their oxidative defense activation and prebiotic activities.

BACKGROUND: Codonopsis pilosula and C. tangshen are both plants widely used in traditional Chinese medicine. Polysaccharides, which are their primary active components, are thought to be important in their extensive use. In this study, two neutral polysaccharide fractions of C. pilosula (CPPN) and C. tangshen (CTPN) were obtained by fractionation on a DEAE-Sepharose column and characterized. RESULTS: It was confirmed that the neutral polymers CPPN and CTPN were ß-(2,1)-linked inulin-type fructans with non-reducing terminal glucose, and degree of polymerization (DP) of 19.6 and 25.2, respectively. The antioxidant and prebiotic activities in vitro were assayed based on IPEC-J2 cell lines and five strains of Lactobacillus. Results indicated that the effects of CPPN and CTPN were increased antioxidant defense in intestinal epithelial cells through enhanced cell viability, improved expression of total antioxidant capacity, glutathione peroxidase, superoxide dismutase and catalase, and reduced levels of malondialdehyde and lactic dehydrogenase. The prebiotic activity of CPPN and CTPN was demonstrated by the promoting effect on Lactobacillus proliferation in vitro. The different biological activities obtained between the two fractions are probably due to the different DP and thus molecular weights of CPPN and CTPN. CONCLUSION: The inulin fractions from C. pilosula and C. tangshen were natural sources of potential intestinal antioxidants as well as prebiotics, which will be valuable in further studies and new applications of inulin-containing health products. © 2020 Society of Chemical Industry.

Extraction of Inulin from Andean Plants: An Approach to Non-Traditional Crops of Ecuador.

Inulin is a polysaccharide of fructose widely used in the food and pharmaceutical industry due to its physicochemical properties and technological applications. Inulin from jicama (Smallanthus sonchifolius) and cabuya (Agave americana) was obtained. The steps for inulin obtention were: raw material preparation, extraction and purification. The extraction conditions were determined using a random design with three levels of stirring speed (0, 130 and 300 rpm), and a 32 factorial experimental design with three levels of temperature (40, 60 and 80 °C) and solid:liquid ratio (1:2, 1:3 and 1:5 S:L). The results showed that the best extractions conditions for jicama were 130 rpm, 75 °C, 1:5 S:L and 25 min; while for cabuya were 80 °C, 300 rpm, 1:5 S-L and 100 min. The weight average molecular weight of inulin from jicama and cabuya were 5799.9 and 4659.75 g/mol, respectively. The identity of the obtained inulin from jicama and cabuya were confirmed by infrared (IR) and Raman spectroscopy. In addition, scanning electron microscopy, differential scanning calorimetry and thermogravimetry analyses were performed to characterize both inulins.

High yield production of levan-type fructans by Gluconobacter japonicus LMG 1417.

Levan, a ß-2,6-glycosidic linked fructan, is a promising alternative for the inulin dominated fructan market. Although levan is already used in some cosmetic products, the commercial availability of the fructan is still limited. Here we show that Gluconobacter japonicus LMG 1417 is a potent levan-forming organism and a promising platform for the industrial production of levan. The levansucrase LevS1417, which is produced by G. japonicus LMG 1417 and secreted by a signal-peptide-independent pathway, exhibited extraordinary high activity (4726 ±â€¯821 U mg-1 at 50 °C). A cell-free levan production based on the supernatant of the investigated strain led to a final levan yield of 157.9 ±â€¯7.6 g L-1. The amount of secreted levansucrase was more than doubled by plasmid-mediated homologous overproduction of LevS1417 in G. japonicus LMG 1417. Accordingly, the space-time yield of cell-free levan production was doubled using the plasmid-bearing mutant.

Molecular weight dependent structure of the exopolysaccharide levan.

Levan is a bacterial homopolysaccharide, which consists of ß-2➔6 linked ß-(D)-fructose monomers. Because of its structural properties and its health promoting effects, levan is a promising functional ingredient for food, cosmetic and pharmaceutical products. The properties of levan have been reported to be linked to its molecular weight. For a better understanding of how the molecular weight determines its polymer conformation in aqueous solution, levan produced by the food grade acetic acid bacterium Gluconobacter albidus TMW 2.1191 was analysed over a broad molecular weight range using dynamic and static light scattering and viscometry. Low molecular weight levan exhibit a compact random coil structure. As the molecular weight increases, the structure transforms into a compact non-drained sphere. The density of the sphere continues to increase with increasing molecular weight. This resulted in a negative exponent in the Mark-Houwink-Sakurada Plot. For the first time, an increase in molecular density with increasing molecular weight, as determined by a negative Mark-Houwink-Sakurada exponent, can be shown for biopolymers. Our results reveal the unique properties of high-molecular weight levan and confirm the need of further systematic studies on the structure-function relationship of levan for its targeted use in food, cosmetic and pharmaceutical applications.

Purification and structural characterization and antioxidant activity of levan from Bacillus megaterium PFY-147.

An exopolysaccharide (EPS)-producing bacterial PFY-147 was separated from Hiqiher vineyard soil and determined as Bacillus megaterium by 16S rDNA analysis and morphological. The PFY-147-EPS was purified by gel-filtration chromatography. The average molecular weight (Mw) of purified EPS was 1.878 × 106 Da. The total carbohydrate contents, uronic acid and sulfated group content in PFY-147-EPS were found to be 90.37 ± 1.48%, 7.24 ± 0.36% and 2.39 ± 0.36%, respectively. The PFY-147-EPS was found to be a levan containing a backbone of 2,6-substituted ß-fructoses by high performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR) and Fourier transform infrared (FT-IR) spectroscopy. The atomic force microscopy (AFM) presented that the levan formed pointed or thorny structural in aqueous solution. Scanning electron microscopy (SEM) of the levan showed compact and nonuniform block sizes morphology. Moreover, the levan exhibited higher thermal stability with a degradation temperature of 265.63 °C in DSC analysis. The levan possessed strong 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, Superoxide anion (O2-) radical, hydroxyl (OH) radical and and2,20-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS+) scavenging activities and strong metal ion chelating activity. The WHC and WSI of levan were 231.29 ± 3.76% and 97.34 ± 1.72%, respectively. These might have implications in food and medicine, as well as in pharmaceutical industries.

Chemical structure elucidation of an inulin-type fructan isolated from Lobelia chinensis lour with anti-obesity activity on diet-induced mice.

A neutral polysaccharide (LCPS) was obtained from Lobelia chinensis via hot water extraction and ethanol precipitation. After separation and purification, the homogeneous polysaccharide was obtained with a molecular weight of 2.6 × 103 Da. The chemical composition of the extracted polysaccharide contains fructose and glucose with protein-free identified by gas chromatography-mass spectrometer (GC-MS) method. Chemical structure of LCPS was indicated by nuclear magnetic resonance (NMR) spectroscopy. Data indicated that LCPS was an inulin-type fructan with α-D-Glcp-(1→[1)-ß-D-Fruf-(2]15 linkage. LCPS intake reduced at significant level high fat diet (HFD)-induced body weight gain, liver weight, total cholesterol and triglyceride in serum and hepatic tissue, respectively. The lipid droplet accumulation in hepatic tissue was similar between lean and LCPS groups lower than in HFD-induced mice from tissue section staining results. Together, polysaccharides from Lobelia chinensis could be a new source of natural anti-obesity agent against obesity with potential value in the manufacturing supplements and drugs.

Levan from a new isolated Bacillus subtilis AF17: Purification, structural analysis and antioxidant activities.

A strain of Bacillus subtilis AF 17 with high exopolysaccharide (EPS) production ability was isolated and identified based on morphological and physiological characteristics and phylogenetic analysis of 16S rDNA sequences. EPS was isolated from the strain fermentation broth by alcohol precipitation and gel-filtration chromatography. Its structural characteristics were investigated and elucidated by methylation analysis, gas chromatography mass spectrometry and nuclear magnetic resonance spectroscopy. Based on the obtained data, the EPS was found to be a levan containing a backbone of 6-substituted ß-fructoses, with a low grade of branching at position 1 (linear/branched ratio 20:1). Levan showed a molecular weight of about 20 MDa. The antioxidant activity of this biopolymer was studied and revealed that levan showed an interesting 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging capacity (IC50 levan = 1.42 mg/mL), reducing power, and also a strong total antioxidant activity. Overall, the results suggest that levan is a promising source of natural antioxidants and can be used as additive in food and pharmaceutical preparations.

Role of levan extracted from bacterial honey isolates in curing peptic ulcer: In vivo.

Peptic ulcer is one of the worldwide diseases where 10% of adults are affected by peptic ulcers at least once in their lifetime. The goal of this study was to evaluate the effect of levan in treating peptic ulcer. The bacterial honey isolates called Bacillus sp. levan was utilized. Levan was chemically characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), 1H and 13C NMR analysis. Levan was used to treat gastric ulcers induced in rats by oral administration of 5 mL/kg ethanol. Microscopic examination of stomach sections indicated that treatment with 200 mg/kg levan effectively healed the ulcers. Levan had no antimicrobial activity against a common cause of ulcers such as Helicobacter pylori bacteria. Rather, we proposed that the high adhesion (manifested as a protective coating) and prebiotic activity of levan may account for the observed beneficial effects. The immunohistochemical examination showed that levan led to a noticeable Bacillus sp. levan reduction in NF-κB in the upper gastric mucosa. The results concluded that the role of levan was more protective rather than preventive and suggested that levan could play a fundamental role in solving the peptic ulcer problems.

Fructans as Immunomodulatory and Antiviral Agents: The Case of Echinacea.

Throughout history, medicinal purposes of plants have been studied, documented, and acknowledged as an integral part of human healthcare systems. The development of modern medicine still relies largely on this historical knowledge of the use and preparation of plants and their extracts. Further research into the human microbiome highlights the interaction between immunomodulatory responses and plant-derived, prebiotic compounds. One such group of compounds includes the inulin-type fructans (ITFs), which may also act as signaling molecules and antioxidants. These multifunctional compounds occur in a small proportion of plants, many of which have recognized medicinal properties. Echinacea is a well-known medicinal plant and products derived from it are sold globally for its cold- and flu-preventative and general health-promoting properties. Despite the well-documented phytochemical profile of Echinacea plants and products, little research has looked into the possible role of ITFs in these products. This review aims to highlight the occurrence of ITFs in Echinacea derived formulations and the potential role they play in immunomodulation.

Extraction and Characterization of Inulin-Type Fructans from Artichoke Wastes and Their Effect on the Growth of Intestinal Bacteria Associated with Health.

Globe artichoke is an intriguing source of indigestible sugar polymers such as inulin-type fructans. In this study, the effect of ultrasound in combination with ethanol precipitation to enhance the extraction of long chain fructans from artichoke wastes has been evaluated. The inulin-type fructans content both from bracts and stems was measured using an enzymatic fructanase-based assay, while its average degree of polymerization (DP) was determined by HPLC-RID analysis. Results show that this method provides artichoke extracts with an inulin-type fructans content of 70% with an average DP between 32 and 42 both in bracts and in stems. The prebiotic effect of long chain inulins from artichoke extract wastes was demonstrated by its ability to support the growth of five Lactobacillus and four Bifidobacterium species, previously characterized as probiotics. Besides, we considered the possibility to industrialize the process developing a simpler method for the production of inulin-type fructans from the artichoke wastes so that the artichoke inulin preparation could be suitable for its use in synbiotic formulations in combination with different probiotics for further studies including in vivo trials.

Clinical Efficacy of Oligofructans from Ophiopogon japonicus in Reducing Atopic Dermatitis Flare-ups in Caucasian Patients.

Atopic dermatitis is a chronic relapsing inflammatory skin disease affecting 15-20% children and 2-10% adults worldwide. Topical treatments include corticosteroids and calcineurin inhibitors, despite frequently observed adverse events such as skin atrophy, itching and burning sensations. Good alternatives that can prolong disease relief in between flare-ups are therefore needed. We conducted a randomized, single-blind, placebo-controlled, multicenter clinical trial in a Caucasian cohort of 90 children and 144 adults with mild-to-moderate atopic dermatitis that applied tested products twice daily for 60 days. A natural active from Ophiopogon japonicus, that improves atopic dermatitis symptoms in vivo, was successful in reducing the SCORing of Atopic Dermatitis (SCORAD), including erythema, pruritus and body surface area in both cohorts. The active also improved patient's quality of life and significantly reduced the number of patients relapsing compared to placebo. We conclude that this treatment could be an effective solution to help control the disease in between flare-ups.

Characterization of Inulin-Type Fructan from Platycodon grandiflorus and Study on Its Prebiotic and Immunomodulating Activity.

Platycodon grandiflorus is a plant widely used in traditional Chinese medicine, of which polysaccharides are reported to be the main components responsible for its bio-functions. In this work, the inulin-type fructan (PGF) was obtained by DEAE anion exchange chromatography from the water extracted from P. grandifloras. Characterization was performed with methanolysis, methylation, and NMR and the results showed that PGF is a ß-(2-1) linked fructan, with terminal glucose and with a degree of polymerization of 2⁻10. In order to study its biofunctions, the prebiotic and immunomodulation properties were assayed. We found that PGF exhibited good prebiotic activity, as shown by a promotion on six strains of lactobacillus proliferation. Additionally, the PGF also displayed direct immunomodulation on intestinal epithelial cells and stimulated the expressions of anti-inflammatory factors. These results indicated that the inulin from P. grandiflorus is a potential natural source of prebiotics as well as a potential intestinal immunomodulator, which will be valuable for further studies and new applications.

Isolation and structural characterization of levan produced by probiotic Bacillus tequilensis-GM from Tunisian fermented goat milk.

The probiotic features of strain GM newly isolated from Tunisian spontaneously fermented goat milk and identified as Bacillus tequilensis-GM were assessed. Strain GM showed high resistance to saliva (90.64%), gastric juice (88.55%), intestinal juice (72.83%) and resistance to bile salts (65.22%), was able to act against Listeria monocytogenes ATCC 15313, Escherichia coli ATCC 25922 and Enterococcus feacalis ATCC 25912, showed high surface hydrophobicity (77.3%) and was sensitive to most of the studied antibiotics. Strain GM did not exhibit any hemolytic activity whereas it was able to produce protease, amylase and ß-galactosidase. Moreover, results showed that strain GM produced high molecular weight ß-(2 → 6)-levan with high ability to inhibit and to disrupt pathogenic biofilms and with high ability to reduce syneresis of sucrose-supplemented skimmed milk. B. tequilensis-GM can therefore be suitable to be used as starter culture in fermented dairy products, since it possesses desirable probiotic properties in addition to its ability to produce levan.

Purification, characterization and immunomodulatory activity of fructans from Polygonatum odoratum and P. cyrtonema.

Two neutral polysaccharides named POP-1 and PCP-1 were purified from Polygonatum odoratum and P. cyrtonema, respectively. Results showed that both of them were composed of fructose and glucose, and the average molecular weights of them were about 5 kDa. FT-IR, 1D- and 2D-NMR analyses revealed that both POP-1 and PCP-1 contained a (2→1)-linked ß-d-fructofuranose (Fruf) backbone and (2→6)-linked ß-d-Fruf side chains with an internal α-d-glucopyranose (Glcp) in neokestose form. The difference between these two purified polysaccharides were that PCP-1 possessed of an acetyl group attached at O-3 of α-d-Glcp residue. In addition, PCP-1 exhibited a little better immune stimulating activity than POP-1 on cell viability and IL-6 production of RAW 264.7 macrophages. These results indicated that the acetyl group might affect the immunoregulatory activity and PCP-1 have more potential to be explored as an immunomodulatory agent.

Recent development trends for chiral stationary phases based on chitosan derivatives, cyclofructan derivatives and chiral porous materials in high performance liquid chromatography.

The separation of enantiomers by chromatographic methods, such as gas chromatography, high-performance liquid chromatography and capillary electrochromatography, has become an increasingly significant challenge over the past few decades due to the demand of pharmaceutical, agrochemical, and food analysis. Among these chromatographic resolution methods, high-performance liquid chromatography based on chiral stationary phases has become the most popular and effective method used for the analytical and preparative separation of optically active compounds. This review mainly focuses on the recent development trends for novel chiral stationary phases based on chitosan derivatives, cyclofructan derivatives, and chiral porous materials that include metal-organic frameworks and covalent organic frameworks in high-performance liquid chromatography. The enantioseparation performance and chiral recognition mechanisms of these newly developed chiral selectors toward enantiomers are discussed in detail.