Biochemical Characteristics and Potential Biomedical Applications of Hydrolyzed Carrageenans.

Seaweed contains a variety of bioactive compounds; the most abundant of them are polysaccharides, which have significant biological and chemical importance. Although algal polysaccharides, especially the sulfated polysaccharides, have great potential in the pharmaceutical, medical and cosmeceutical sectors, the large molecular size often limits their industrial applications. The current study aims to determine the bioactivities of degraded red algal polysaccharides by several in vitro experiments. The molecular weight was determined by size-exclusion chromatography (SEC), and the structure was confirmed by FTIR and NMR. In comparison to the original furcellaran, the furcellaran with lower molecular weight had higher OH scavenging activities. The reduction in molecular weight of the sulfated polysaccharides resulted in a significant decrease in anticoagulant activities. Tyrosinase inhibition improved 2.5 times for hydrolyzed furcellaran. The alamarBlue assay was used to determine the effects of different Mw of furcellaran, κ-carrageenan and ι-carrageenan on the cell viability of RAW264.7, HDF and HaCaT cell lines. It was found that hydrolyzed κ-carrageenan and ι-carrageenan enhanced cell proliferation and improved wound healing, whereas hydrolyzed furcellaran did not affect cell proliferation in any of the cell lines. Nitric oxide (NO) production decreased sequentially as the Mw of the polysaccharides decreased, which indicates that hydrolyzed κ-Carrageenan, ι-carrageenan and furcellaran have the potential to treat inflammatory disease. These findings suggested that the bioactivities of polysaccharides were highly dependent on their Mw, and the hydrolyzed carrageenans could be used in new drug development as well as cosmeceutical applications.

Brewing of glucuronic acid-enriched apple cider with enhanced antioxidant activities through the co-fermentation of yeast (Saccharomyces cerevisiae and Pichia kudriavzevii) and bacteria (Lactobacillus plantarum).

Co-fermentation using yeast (Saccharomyces cerevisiae and Pichia kudriavzevii) and the bacteria (Lactobacillus plantarum) as starters isolated from spontaneous sourdough was conducted for the brewing of glucuronic acid (GlcA)-enriched apple cider. The concentration of GlcA in the apple cider co-fermented for 14 d with commercial S. cerevisiae and L. plantarum was 37.7 ± 1.7 mg/mL while a concentration of 62.8 ± 3.1 mg/mL was recorded for fermentation with P. kudriavzevii and L. plantarum, which was higher than the corresponding single yeast fermentation. The co-fermented apple cider revealed higher 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of 171.67 ± 0.79 µg trolox equivalents (TE)/mL using P. kudriavzevii and L. plantarum, compared to the control (143.89 ± 7.07 µg TE/mL) just using S. cerevisiae. Thus, the co-fermentation of S. cerevisiae and L. plantarum and P. kudriavzevii and L. plantarum provided a new strategy for the development of GlcA-enriched apple cider with enhanced antioxidant capacity.

Characteristics of sourdough bread fermented with Pediococcus pentosaceus and Saccharomyces cerevisiae and its bio-preservative effect against Aspergillus flavus.

Six lactic acid bacteria (LAB) and four yeast strains were isolated from Pyeongchang spontaneous sourdough. In combination with the segregated Saccharomycopsis fibuligera and Saccharomyces cerevisiae, Pediococcus pentosaceus was employed for sourdough bread starters because of its antifungal action against Aspergillus flavus. The sourdough bread fermented with P. pentosaceus and S. cerevisiae displayed 56.4% ± 5.5% antifungal movement counter to A. flavus expansion at 96 h. The concentration of lactic and acetic acids in the sourdough bread was 4.5- and 1.6-folds above the control bread, respectively, contributing to the balanced sensory properties with a fermentation quotient (FQ) of 2.08-2.86. SPME- GC/MS newly distinguished twenty-two volatile compounds including six aldehydes, five alcohols, one phenol, three ketones, one acid, and six esters. The results suggest the P. pentosaceus and S. cerevisiae combination as promising sourdough starters for making enhanced quality bread free of preservatives.

Synthesis and biological evaluation of chalcone derivatives as neuroprotective agents against glutamate-induced HT22 mouse hippocampal neuronal cell death.

Seventeen chalcone analogues were synthesized from 7-methoxy-3,4-dihydronaphthalen1(2H)-one and various aromatic aldehydes under basic conditions and their therapeutic properties were studied in mouse hippocampal cell line HT-22 against neuronal cell death induced by glutamate. From this study, we selected an analogue C01 as a active compound which showed significantly high neuroprotection. This compound inhibited Ca2+ influx and reactive oxygen species (ROS) accumulation inside cells. The glutamate-induced cell death was analyzed by flow cytometry and it showed that C01 significantly reduced apoptotic or dead cell induced by 5 mM glutamate. Western blot analysis indicates that glutamate-mediated activation of MAPKs were inhibited by compound C01 treatment. In addition, the C01enhanced Bcl-2 and decreased Bax, the anti and pro apoptotic proteins respectively. Further analysis showed that, C01 prevented the nuclear translocation of AIF (apoptosis inducing factor) and inhibited neuronal cell death. Taken together, compound C01 treatment resulted in decreased neurotoxicity induced by 5 mM of glutamate. Our finding confirmed that compound C01 has neuro-therapeutic potential against glutamate-mediated neurotoxicity.

Enhancement of neuroprotection, antioxidant capacity, and water-solubility of crocins by transglucosylation using dextransucrase under high hydrostatic pressure.

Crocin, one of the major carotenoid pigments of Crocus sativus (saffron), is responsible for antioxidant activity, neuroprotection, and the inhibition of tumor cell proliferation. In order to improve the functionality of crocin, α-glucosyl-(1→6)-trans-crocins (C-Gs) were synthesized using sucrose and dextransucrase from Leuconostoc mesenteroides. High hydrostatic pressure (HHP) technique was applied to the synthesis process of C-Gs in order to improve its transglucosylation yield. A 100 MPa HHP condition enhanced the production yield of C-Gs by 1.95 times compared to that of 0.1 MPa atmospheric pressure. Novel C-Gs were purified by HPLC, and their chemical structures were determined using NMR analysis. Novel C-Gs increased water solubility 4.6-5.7 times and antioxidant activity 1.5-2.6 times, respectively, compared to crocin, and their neuroprotections (cell viability 92.5-100.4 %) on HT22 mouse hippocampal neuronal cells were significantly higher than that of crocin (cell viability 84.6 %). This advanced neuroprotection of novel C-Gs could be highly associated with their enhanced antioxidant activity. Thus, the enhanced water solubility and functionality of novel C-Gs can induce better clinical efficacy of neuroprotection than trans-crocin.

Fermented Wild Ginseng by Rhizopus oligosporus Improved l-Carnitine and Ginsenoside Contents.

We conducted this study to investigate the beneficial effects of Rhizopus oligosporus fermentation of wild ginseng on ginsenosides, l-carnitine contents and its biological activity. The Rhizopus oligosporus fermentation of wild ginseng was carried out at 30 °C for between 1 and 14 days. Fourteen ginsenosides and l-carnitine were analyzed in the fermented wild ginseng by the ultra high pressure liquid chromatography-mass spectrometry (UPLC-MS) system. Our results showed that the total amount of ginsenosides in ginseng increased from 3,274 to 5,573 mg/kg after 14 days of fermentation. Among the 14 ginsenosides tested, the amounts of 13 ginsenosides (Rg1, Rb2, Rb3, Rc, Rd, Re, Rf, Rg2, Rg3, Rh1, compound K, F1 and F2) increased, whereas ginsenoside Rb1 decreased, during the fermentation. Furthermore, l-carnitine (630 mg/kg) was newly synthesized in fermented ginseng extract after 14 days. In addition, both total phenol contents and DPPH radical scavenging activities showed an increase in the fermented ginseng with respect to non-fermented ginseng. These results show that the fermentation process reduced the cytotoxicity of wild ginseng against RAW264.7 cells. Both wild and fermented wild ginseng showed anti-inflammatory activity via inhibition of nitric oxide synthesis in RAW264.7 murine macrophage cells.

Composition and biochemical properties of ale beer enriched with lignans from Schisandra chinensis Baillon (omija) fruits.

To develop a beverage with high antioxidant capacity and desirable sensory characteristics, Schisandra chinensis (omija) fruits were added to ale type beer at different time points of the brewing process. The phenolic compounds contents in beer were found to be dependent at the moment of the addition of omija fruit. Addition of omija fruits at the initiation of boiling imparted highest oxidative stability to beer and resulted in highest total phenolic and flavonoid contents in ale beer (606.82 mg GAE/L and 406.75 mg QE/L, respectively). The amounts of schisandrin, gomisin A and gomisin B in beer were 12.10 mg/mL, 3.12 mg/mL and 0.86 mg/mL, respectively. Taken together, it is hypothesized that the addition of omija fruits to traditional brewing process can improve the development of value-added beer products.

Anti-cariogenic Characteristics of Rubusoside.

Streptococcus mutans plays an important role in the development of dental caries in humans by synthesizing adhesive insoluble glucans from sucrose by mutansucrase activity. To explore the anti-cariogenic characteristics of rubusoside (Ru), a natural sweetener component in Rubus suavissimus S. Lee (Rosaceae), we investigated the inhibitory effect of Ru against the activity of mutansucrase and the growth of Streptococcus mutans. Ru (50 mM) showed 97% inhibitory activity against 0.1 U/mL mutansucrase of S. mutans with 500 mM sucrose. It showed competitive inhibition with a K i value of 1.1 ± 0.2 mM and IC50 of 2.3 mM. Its inhibition activity was due to hydrophobic and hydrogen bonding interactions based on molecular docking analysis. Ru inhibited the growth of S. mutans as a bacteriostatic agent, with MIC and MBC values of 6 mM and 8 mM, respectively. In addition, Ru showed synergistic anti-bacterial activity when it was combined with curcumin. Therefore, Ru is a natural anti-cariogenic agent with anti-mutansucrase activity and antimicrobial activity against S. mutans. ELECTRONIC SUPPLEMENTARY MATERIAL ESM: The online version of this article (doi: 10.1007/s12257-018-0408-0) contains supplementary material, which is available to authorized users.

Fucoxanthin bioavailability from fucoxanthin-fortified milk: In vivo and in vitro study.

Our previous study reported the improved stability of fucoxanthin (FX) fortified in whole milk (WM) and skimmed milk (SM). In this study, in vivo and in vitro FX bioavailability were investigated using FX-fortified milk (FX-SM and FX-WM) and microalga Phaeodactylum tricornutum biomass (Pt-powder). Organ tissue accumulation of FX and its metabolites (FXOH: fucoxanthinol, AXA: amarouciaxanthin A) after repeated oral administration was in the following order: FX-SM > FX-WM > Pt-powder. In vivo pharmacokinetic study with a single oral administration also demonstrated that the absorption of FXOH and AXA was the highest for FX-SM. To reinforce the in vivo results, in vitro-simulated digestion and Caco-2 cell uptake assays were performed, which revealed that FX-SM showed the highest FX bioaccessibility (release from food matrices) and cellular uptake efficiency of FX and FXOH. In conclusion, skimmed milk was validated as an excellent food matrix for FX application in terms of stability and bioavailability.

Decrease of insoluble glucan formation in Streptococcus mutans by co-cultivation with Enterococcus faecium T7 and glucanase addition.

OBJECTIVES: To develop preventive canine oral health bio-materials consisting of probiotics and glucanase to reduce insoluble glucan and volatile sulfur compound formation. RESULTS: Co-cultivation of Enterococcus faecium T7 with Streptococcus mutans at inoculation ratio of 3:1 (v/v) resulted in 25% reduction in the growth of Streptococcus mutans. Amounts of soluble and insoluble glucans produced by S. mutans were decreased to 70 and 55%, respectively. Insoluble glucan was decreased from 0.6 µg/ml in S. mutans culture to 0.03 µg/ml in S. mutans co-cultivated with E. faecium T7 in the presence of Lipomyces starkeyi glucanase. Volatile sulfur compound, a main component of halitosis produced by Fusobacteria nucleatum, was decreased by co-cultivating F. nucleatum with E. faecium. CONCLUSION: E. faecium and glucanase can be combined as potentially active ingredients of oral care products for pets by reducing plaque-forming bacteria growth and their by-products that cause cavity and periodontal disease.

Preparation of Fucoxanthin-Loaded Nanoparticles Composed of Casein and Chitosan with Improved Fucoxanthin Bioavailability.

To facilitate the utilization of fucoxanthin (FX), a valuable marine carotenoid, in the food industry, FX-loaded casein nanoparticles (FX-CN) and chitosan-coated FX-CN (FX-CS-CN) were developed using the FX-enriched fraction from Phaeodactylum tricornutum. Two nanoscale particles (237 ± 13 nm for FX-CN and 277 ± 26 nm for FX-CN-CN) with spherical and smooth surfaces showed over 71% encapsulation efficiency and polydispersity index (PDI) value of 0.31-0.39 in water. Owing to the chitosan coating, FX-CS-CN showed a positive zeta potential (24.00 mV), whereas that of FX-CN was negative (-12.87 mV). In vitro simulated digestion demonstrated better FX bioaccessibility from the nanoparticles versus P. tricornutum powder (Pt-powder) and from FX-CN versus FX-CS-CN. However, in C57BL/6 mice, fucoxanthinol absorption to the blood circulation was two times higher for FX-CS-CN versus FX-CN, possibly due to increased retention or adsorption to mucin by the cationic biopolymer in the chitosan-coated particles. These results demonstrate that FX-CS-CN can enable the application of FX, with improved bioavailability and water dispersibility, in the food industry.

Development, Quantification, Method Validation, and Stability Study of a Novel Fucoxanthin-Fortified Milk.

To extend the scope of application of fucoxanthin, a marine carotenoid, whole milk (WM) and skimmed milk (SM) were fortified with fucoxanthin isolated from the microalga Phaeodactylum tricornutum to a final 8 µg/mL milk solution concentration. Using these liquid systems, a fucoxanthin analysis method implementing extraction and HPLC-DAD was developed and validated by accuracy, precision, system suitability, and robustness tests. The current method demonstrated good linearity over the range of 0.125-100 µg/mL fucoxanthin with R(2) = 1.0000, and all validation data supported its adequacy for use in fucoxanthin analysis from milk solution. To investigate fucoxanthin stability during milk production and distribution, fucoxanthin content was examined during storage, pasteurization, and drying processes under various conditions. Fucoxanthin in milk solutions showed better stabilizing effect in 1 month of storage period. Degradation rate constant (k) on fucoxanthin during this storage period suggested that fucoxanthin stability might be negatively correlated with decrease of temperature and increase of protein content such as casein and whey protein in milk matrix. In a comparison between SM and WM, fucoxantin in SM always showed better stability than that in WM during storage and three kinds of drying processes. This effect was also deduced to relate with protein content. In the pasteurization step, >91% of fucoxanthin was retained after three pasteurization processes even though the above trend was not found. This study demonstrated for the first time that milk products can be used as a basic food matrix for fucoxanthin application and that protein content in milk is an important factor for fucoxanthin stability.