Structural characteristics and functional properties of a fucose containing prebiotic exopolysaccharide from Bifidobacterium breve NCIM 5671.

AIM: To evaluate the structure and functions of capsular exopolysaccharide (CPS) from Bifidobacterium breve NCIM 5671. METHODS AND RESULTS: A CPS produced by the probiotic bacteria B. breve NCIM 5671 was isolated and subjected to characterization through GC analysis, which indicated the presence of rhamnose, fucose, galactose, and glucose in a molar ratio of 3:1:5:3. The average molecular weight of the CPS was determined to be ∼8.5 × 105 Da. Further, NMR analysis revealed the probable CPS structure to be composed of major branched tetra- and penta-saccharide units alternately repeating and having both α- and ß-configuration sugar residues. CPS displayed an encouraging prebiotic score for some of the studied probiotic bacteria. Compared to standard inulin, CPS showed better resistance to digestibility against human GI tract in vitro. DPPH, total antioxidant, and ferric reducing assays carried out for CPS displayed decent antioxidant activity too. CONCLUSION: This study indicates that the CPS from B. breve NCIM 5671 has the potential to be utilized as a prebiotic food supplement. It is a high-molecular-weight (∼8.5 × 105 Da) capsular heteropolysaccharide containing rhamnose, fucose, galactose, and glucose.

Preventive and therapeutic aspects of fermented foods.

In recent times, the status of some fermented foods which are considered as functional foods that confer health benefits in certain disease conditions has grown rapidly. The health benefits of fermented foods are due to the presence of probiotic microbes and the bioactive compounds formed during fermentation. Microbes involved and metabolites produced by them are highly species specific and contribute to the authenticity of the fermented foods. Several studies pertaining to the effect of fermented foods on various disease conditions have been conducted in recent years using both animal models and clinical trials on humans. This review focuses on the impact of fermented foods on conditions such as diabetes, cardiovascular disease, obesity, gastrointestinal disorder, cancer and neurodegenerative disorders.

Prebiotic Chondroitin Sulfate Disaccharide Isolated from Chicken Keel Bone Exhibiting Anticancer Potential Against Human Colon Cancer Cells.

Chondroitin sulfate (CS)-Keel disaccharide (CSD) was produced by chondroitin AC lyase (PsPL8A) degradation of food grade CS-Keel polysaccharide isolated from chicken keel cartilage. PsPL8A showed specific activity, 340 ± 5.2 U mg-1 with CS-Keel polysaccharide. TLC showed CSD as the major product. CSD was purified by gel filtration and MS/MS, confirms it as C4S disaccharide. Structural characterization by FTIR and NMR showed presence of N-acetylgalactosamine and glucuronic acid. CSD displayed resistance to gastric juice with 23.7% hydrolysis at pH 1.0. CSD showed prebiotic score of 0.57 for L. acidophilus and 0.58 for B. infantis and produce SCFA products. MTT and morphological analysis confirmed, CSD (0.5 mg mL-1) does not decrease viability of mouse fibroblast (L929), but showed anti-proliferative potential against human colon cancer (HT-29) cell lines (80% inhibition). CSD treated HT-29 cells exhibit nuclear fragmentation and apoptosis. Prebiotic and anticancer potential of CSD can be utilized for functional food preparation and prevention of gastrointestinal disorders.

Structure modeling and functional analysis of recombinant dextransucrase from Weissella confusa Cab3 expressed in Lactococcus lactis.

The dextransucrase gene from Weissella confusa Cab3, having an open reading frame of 4.2 kb coding for 1,402 amino acids, was amplified, cloned, and expressed in Lactococcus lactis. The recombinant dextransucrase, WcCab3-rDSR was expressed as extracellular enzyme in M17 medium with a specific activity of 1.5 U/mg which after purification by PEG-400 fractionation gave 6.1 U/mg resulting in 4-fold purification. WcCab3-rDSR was expressed as soluble and homogeneous protein of molecular mass, approximately, 180 kDa as analyzed by SDS-PAGE. It displayed maximum enzyme activity at 35°C at pH 5.0 in 50 mM sodium acetate buffer. WcCab3-rDSR gave Km of 6.2 mM and Vm of 6.3 µmol/min/mg. The characterization of dextran synthesized by WcCab3-rDSR by Fourier transform infrared and nuclear magnetic resonance spectroscopic analyses revealed the structural similarities with the dextran produced by the native dextransucrase. The modeled structure of WcCab3-rDSR using the crystal structures of dextransucrase from Lactobacillus reuteri (protein data bank, PDB id: 3HZ3) and Streptococcus mutans (PDB id: 3AIB) as templates depicted the presence of different domains such as A, B, C, IV, and V. The domains A and B are circularly permuted in nature having (ß/α)8 triose phosphate isomerase-barrel fold making the catalytic core of WcCab3-rDSR. The structure superposition and multiple sequence alignment analyses of WcCab3-rDSR with available structures of enzymes from family 70 GH suggested that the amino acid residue Asp510 acts as a nucleophile, Glu548 acts as a catalytic acid/base, whereas Asp621 acts as a transition-state stabilizer and these residues are found to be conserved within the family.

Immobilization of glucansucrase for the production of gluco-oligosaccharides from Leuconostoc mesenteroides.

Glucansucrase from Leuconostoc mesenteroides was immobilized in 1 % (w/v) with sodium alginate to produce oligosaccharides. Glucansucrase gave three activity bands of approx. 240, 178, and 165 kDa after periodic acid-Schiff staining with sucrose. The immobilized enzyme had 40 % activity after ten batch reactions at 30 °C and 75 % activity after a month of storage at 4 °C, which is six times more stable than the free enzyme. Immobilized enzyme was more stable at lower (3.5-4.5) and higher (6.5-7.0) pH ranges and higher temperatures (35-40 °C) compared with the free enzyme. Immobilized and free glucansucrase were employed in the acceptor reaction with maltose and each produced gluco-oligosaccharide ranging from trisaccharides to homologous pentasaccharides.

Dextran Utilization During Its Synthesis by Weissella cibaria RBA12 Can Be Overcome by Fed-Batch Fermentation in a Bioreactor.

Weissella cibaria RBA12 produced a maximum of 9 mg/ml dextran (with 90% efficiency) using shake flask culture under the optimized concentration of medium components viz. 2% (w/v) of each sucrose, yeast extract, and K2HPO4 after incubation at optimized conditions of 20 °C and 180 rpm for 24 h. The optimized medium and conditions were used for scale-up of dextran production from Weissella cibaria RBA12 in 2.5-l working volume under batch fermentation in a bioreactor that yielded a maximum of 9.3 mg/ml dextran (with 93% efficiency) at 14 h. After 14 h, dextran produced was utilized by the bacterium till 18 h in its stationary phase under sucrose depleted conditions. Dextran utilization was further studied by fed-batch fermentation using sucrose feed. Dextran on production under fed-batch fermentation in bioreactor gave 35.8 mg/ml after 32 h. In fed-batch mode, there was no decrease in dextran concentration as observed in the batch mode. This showed that the utilization of dextran by Weissella cibaria RBA12 is initiated when there is sucrose depletion and therefore the presence of sucrose can possibly overcome the dextran hydrolysis. This is the first report of utilization of dextran, post-sucrose depletion by Weissella sp. studied in bioreactor.

Physicochemical, antioxidant and biocompatible properties of chondroitin sulphate isolated from chicken keel bone for potential biomedical applications.

Chicken keel bone cartilage was explored for cheaper and sustainable source for isolation of chondroitin sulphate (CS) for its future use in tissue engineering and pharmaceutical industry. HPSEC analysis displayed two peaks of 100kDa for CS-keel polysaccharide and 1kDa for protein. DLS analysis of CS-keel displayed polydispersity. CS-keel yield was 15% and 53±5% uronic acid content. The quantified percentages of UA-GalNAc4S and UA-GalNAc6S disaccharide in CS-keel were 58% and 42%, respectively. FT-IR identified CS-keel to be chondroitin 4-sulphate. 1H NMR of CS-keel confirmed the presence of N-acetylgalactosamine and Glucuronic acid. FESEM demonstrated layer structure and AFM displayed the size of CS-keel fibres. DSC, TGA and DTG studies of CS-keel showed Td at 243°C. In vitro cell proliferation assay and morphological analysis of mouse fibroblast L929 cell lines confirmed the biocompatibility of CS-keel. CS-keel (5mg/ml) exhibited ∼49% antioxidant activity against DPPH and 22% against superoxide radical protecting from oxidative damage. CS-keel demonstrated better (70.3%) emulsifying activity than commercial sodium alginate (60.2%).

Functional food applications of dextran from Weissella cibaria RBA12 from pummelo (Citrus maxima).

Weissella cibaria RBA12 isolated from pummelo from Northeast India produces a dextran composed of 97% α-(1→6) linkages in the main chain and 3% α-(1→3) branched linkages. The in vitro prebiotic activity of dextran-RBA12 was explored. Dextran-RBA12 displayed enhanced growth of probiotic Bifidobacterium and Lactobacillus spp., and controlled growth of non-probiotic enteric bacteria. Dextran-RBA12 showed superior resistance to physiological barriers with a maximum hydrolysis of 0.51%, 0.31% and 0.24% by artificial gastric juice, α-amylase and intestinal fluid, respectively, whereas compared to maximum hydrolysis of 25.23%, 19.13% and 6%, respectively after 5h of incubation shown by commercial prebiotic inulin. The production of dextran from Weissella cibaria RBA12 in sourdough prepared from whole wheat flour, wheat bran and rye bran showed the highest dextran of 3.26±0.12% d.w. in rye bran. The overall study summarized that dextran-RBA12 can be used as a prebiotic and also can be easily produced in sourdough.

A food additive with prebiotic properties of an α-d-glucan from lactobacillus plantarum DM5.

An α-d-glucan produced by Lactobacillus plantarum DM5 was explored for in vitro prebiotic activities. Glucan-DM5 demonstrated 21.6% solubility, 316.9% water holding capacity, 86.2% flocculation activity, 71.4% emulsification activity and a degradation temperature (Td) of 292.2°C. Glucan-DM5 exhibited lowest digestibility of 0.54% by artificial gastric juice, 0.21% by intestinal fluid and 0.32% by α-amylase whereas the standard prebiotic inulin, showed 25.23%, 5.97% and 19.13%, hydrolysis, respectively. Prebiotic activity assay of glucan-DM5 displayed increased growth of probiotic bacteria such as Bifidobacterium infantis and Lactobacillus acidophilus, but did not support the growth of non-probiotic bacteria such as Escherichia coli and Enterobacter aerogenes. The overall findings indicated that glucan from L. plantarum DM5 can serve as a potential prebiotic additive for food products.