Alkali-assisted extraction, characterization and encapsulation functionality of enzymatic hydrolysis-resistant prolamin from distilled spirit spent grain.

Curcumin is a natural lipophilic polyphenol that exhibits significant various biological properties such as antioxidant and anti-inflammatory properties following oral administration. However, its uses have shown limitations concerning aqueous solubility, bioavailability and biodegradability that could be improved by prolamin-based nanoparticle. In this study, curcumin was encapsulated into prolamin from sorghum (SOP) and wheat (WHP) and distilled spirit spent grain (DSSGP), which was obtained after microbial proteolysis of the former two cereal grains. All the three prolamins showed clear variation of protein profiles and microstructure as confirmed by electrophoresis analysis, disulfide bond determination and Fourier-transform infrared spectroscopy (FTIR). For curcumin-loaded nanospheres (NPs) fabrication, three prolamin-based NPs shared features of spherical shape, uniform particle size, and smooth surface. The average size ranged from 122 to 193 nm depending on the prolamin variety and curcumin loading. In the experiments in vitro, curcumin showed significantly improved UV/thermal stability. Furthermore, DSSGP was more resistant to enzymatic digestion in vitro, hence achieving the controlled release of curcumin in gastrointestinal tract. Collectively, the results indicated the improved bioavailability and biodegradability of curcumin encapsulated by DSSGP, which would be an innovative potential encapsulant for effective protection and targeted delivery of hydrophobic compounds.

Widely targeted metabolomics-based analysis of the impact of L. plantarum and L. paracasei fermentation on rosa roxburghii Tratt juice.

Rosa roxburghii Tratt fruits (RRT) exhibit extremely high nutritional and medicinal properties due to its unique phytochemical composition. Probiotic fermentation is a common method of processing fruits. Variations in the non-volatile metabolites and bioactivities of RRT juice caused by different lactobacilli are not well understood. Therefore, we aimed to profile the non-volatile components and investigate the impact of L. plantarum fermentation (LP) and L. paracasei fermentation (LC) on RRT juice (the control, CG). There were both similarities and differences in the effects of LP and LC on RRT juice. Both of the two strains significantly increased the content of total phenolic, total flavonoid, and some bioactive compounds such as 2-hydroxyisocaproic acid, hydroxytyrosol and indole-3-lactic acid in RRT juice. Interestingly, compared with L. paracasei, L. plantarum showed better ability to increase the content of total phenolic and these valuable compounds, as well as certain bioactivities. The antioxidant capacity and α-glucosidase inhibitory activity of RRT juice were notably enhanced after the fermentations, whereas its cholesterol esterase inhibitory activity was reduced significantly. Moreover, a total of 1466 metabolites were identified in the unfermented and fermented RRT juices. There were 278, 251 and 134 differential metabolites in LP vs CG, LC vs CG, LC vs LP, respectively, most of which were upregulated. The key differential metabolites were classified into amino acids and their derivatives, organic acids, nucleotides and their analogues, phenolic acids and alkaloids, which can serve as potential markers for authentication and discrimination between the unfermented and lactobacilli fermented RRT juice samples. The KEGG enrichment analysis uncovered that metabolic pathways, purine metabolism, nucleotide metabolism and ABC transporters contributed mainly to the formation of unique composition of fermented RRT juice. These results provide good coverage of the metabolome of RRT juice in both unfermented and fermented forms and also provide a reference for future research on the processing of RRT or other fruits.

Characterization and prebiotic potential of polysaccharides from Rosa roxburghii Tratt pomace by ultrasound-assisted extraction.

In this study, polysaccharides (RRTPs) were extracted from Rosa roxburghii Tratt pomace by hot water or ultrasound (US)-assisted extraction. The structural properties and potential prebiotic functions of RRTPs were investigated. Structural characterization was conducted through HPAEC, HPGPC, GC-MS, FT-IR and SEM. Chemical composition analysis revealed that RRTPs extracted by hot water (RRTP-HW) or US with shorter (RRTP-US-S) or longer duration (RRTP-US-L) all consisted of galacturonic acid, galactose, glucose, arabinose, rhamnose and glucuronic acid in various molar ratio. US extraction caused notable reduction in molecular weight of RRTPs but no significant changes in primary structures. Fecal fermentation showed RRTPs could reshape microbial composition toward a healthier balance, leading to a higher production of beneficial metabolites including total short-chain fatty acids, curcumin, noopept, spermidine, 3-feruloylquinic acid and citrulline. More beneficial shifts in bacterial population were observed in RRTP-HW group, while RRTP-US-S had stronger ability to stimulate bacterial short-chain fatty acids production. Additionally, metabolic profiles with the intervention of RRTP-HW, RRTP-US-S or RRTP-US-L were significantly different from each other. The results suggested RRTPs had potential prebiotic effects which could be modified by power US via molecular weight degradation.

Lentinula edodes Sing Polysaccharide: Extraction, Characterization, Bioactivities, and Emulsifying Applications.

In the present work, the optimization of extraction, emulsifying properties, and biological activities of polysaccharides from Lentinula edodes Sing (LES) were studied. The results showed LES polysaccharides extracted by hot water or ultrasonication are a group of ß-glucan. Among all the samples, the one extracted by hot water showed the best emulsifying capacity. In addition, the results demonstrated that LES polysaccharide had strong scavenging activities in vitro on DPPH and ABTS radicals, which reached the highest level for the one extracted by 90 min ultrasonication (p < 0.05). Overall, Lentinula edodes Sing polysaccharides (LESPs) may have potential applications as emulsifying agents in food industries.

Extraction, Characterization, and Platelet Inhibitory Effects of Two Polysaccharides from the Cs-4 Fungus.

Cardiovascular diseases are associated with platelet hyperactivity, and downregulating platelet activation is one of the promising antithrombotic strategies. This study newly extracted two polysaccharides (purified exopolysaccharides, EPSp and purified intercellular exopolysaccharides, IPSp) from Cordyceps sinensis Cs-4 mycelial fermentation powder, and investigated the effects of the two polysaccharides and their gut bacterial metabolites on platelet functions and thrombus formation. EPSp and IPSp are majorly composed of galactose, mannose, glucose, and arabinose. Both EPSp and IPSp mainly contain 4-Galp and 4-Glcp glycosidic linkages. EPSp and IPSp significantly inhibited human platelet activation and aggregation with a dose-dependent manner, and attenuated thrombus formation in mice without increasing bleeding risk. Furthermore, the EPSp and IPSp after fecal fermentation showed enhanced platelet inhibitory effects. The results have demonstrated the potential value of Cs-4 polysaccharides as novel protective ingredients for cardiovascular diseases.

Modification and enhanced anti-inflammatory activity by Bifidobacterial fermentation of an exopolysaccharide from a medicinal fungus Cs-HK1.

The exopolysaccharide (EPS) from the mycelial fermentation of a medicinal fungus Cordyceps sinensis Cs-HK1 had shown significant anti-inflammatory activity previously, and EPS-LM was a highly active fraction with a relatively low molecular weight (MW) isolated from the Cs-HK1 EPS. This study was to assess the effects of Bifidobacterial fermentation in anaerobic conditions on the molecular properties and anti-inflammatory activity of EPS-LM. In both Bifidobacterial cultures (B. breve and B. longum), EPS-LM was fractionally consumed as a carbon source, increasing the bacterial growth and acetic acid production. Analytical results from the fermentation digesta (supernatant) suggested that EPS-LM was partially degraded to lower molecular weight (MW) products with modified structures during the Bifidobacterial fermentation. More interestingly, the higher MW digesta fraction containing the partially degraded EPS-LM showed even stronger inhibiting activity than the original EPS-LM on the LPS-induced pro-inflammatory responses in THP-1 cell culture, including NF-κB activation, release of NO, TNF-α and IL-8. The study has shown that the fermentation by selected Bifidobacterial strains is effective to modify natural polysaccharides with enhanced bioactivities.

Isolation and Assessment of a Highly-Active Anti-Inflammatory Exopolysaccharide from Mycelial Fermentation of a Medicinal Fungus Cs-HK1.

The purpose of this work was to fractionate the complex exopolysaccharide (EPS) from a medicinal fungus Ophiocordyceps sinensis Cs-HK1 based on the molecular weight (MW) range and to assess the in vitro anti-inflammatory activity of different EPS fractions in THP-1 cell culture. The lower MW fraction (EPS-LM-1) showed a much higher anti-inflammatory activity. EPS-LM-1 was identified as a heteropolysaccharide consisting of mannose, glucose, and galactose residues with an average MW of 360 kDa. EPS-LM-1 significantly inhibited the lipopolysaccharide-induced inflammatory responses with the effective concentrations for 50% inhibition below 5 µg/mL on a few major proinflammatory markers. With such a notable in vitro anti-inflammatory activity, EPS-LM-1 is a promising candidate for the development of a new anti-inflammation therapy.

Mechanistic insights into the structure-dependant and strain-specific utilization of wheat arabinoxylan by Bifidobacterium longum.

Arabinoxylan (AX), an important dietary fiber from cereal grains, is mainly metabolised in the large intestine by gut bacteria, especially bifidobacteria. This study investigated the uptake and metabolism of wheat AX by a Bifidobacterium longum strain that could grow well with AX as the sole carbon source. The bacterial growth rate showed a significant correlation to the molecular weight (MW) of AX and its acid hydrolysates. Assessment of the key AX degrading enzymes suggested that the uptake and consumption of AX involved extracellular cleavage of xylan backbone and intracellular degradation of both the backbone and the arabinose substitution. The preference for native or partially hydrolysed AX with single substitutions and a sufficiently high MW suggested the structure-dependant uptake by the bacterial cells. Genetic analysis of B. longum showed the lack of ß-xylosidase, suggesting the existence of unknown enzymes or dual/multiple-specific enzymes for hydrolysis of the non-reducing end of xylan backbone.

A high-molecular weight exopolysaccharide from the Cs-HK1 fungus: Ultrasonic degradation, characterization and in vitro fecal fermentation.

This work was to examine the impact of power ultrasound (US) on the molecular properties of a high-molecular weight (MW) exopolysaccharide (EPS) from the Cs-HK1 medicinal fungus and the utilization, and prebiotic function of the US-treated EPS fractions in human fecal microflora in vitro. The US treatment caused notable reduction of intrinsic viscosity, average MW and aggregate size of EPS in water but no significant changes in the molecular structure. The US-treated EPS fractions were consumed more rapidly by the fecal microflora, resulting in a higher total level of short chain fatty acids. They also affected the relative abundance in the microflora more beneficially than the original EPS. The results suggest that power US is effective for modifying and improving the prebiotic properties of high-MW polysaccharides.

Anti-inflammation activity of exopolysaccharides produced by a medicinal fungus Cordyceps sinensis Cs-HK1 in cell and animal models.

This study was to assess the anti-inflammatory potential of exopolysaccharide (EPS) produced by a medicinal fungus Cordyceps sinensis Cs-HK1. The EPS was isolated from the Cs-HK1 mycelial fermentation broth by ethanol precipitation and purified by deproteinization and dialysis. The EPS had a total sugar content of 74.8% and a maximum average molecular weight (MW) over 107 Da, and consisted mainly of glucose and mannose, and a small amount of galactose and ribose. In THP-1 and RAW264.7 cell cultures, EPS significantly inhibited lipopolysaccharide (LPS)-induced inflammatory responses of the cells including the release of NF-κB and several pro-inflammatory factors such as NO, TNF-α and IL-1ß. In the murine model of LPS-induced acute intestinal injury, the oral administration of EPS to the animals effectively suppressed the expression of major inflammatory cytokines TNF-α, IL-1ß, IL-10 and iNOS and alleviated the intestinal injury. The results suggest that the Cs-HK1 EPS has notable anti-inflammatory activity and can be a potential candidate for further development of new anti-septic therapeutics. To the best of our knowledge, this is the first report on the anti-inflammation of an EPS from C. sinensis fungal fermentation.

Effects of exopolysaccharide fractions with different molecular weights and compositions on fecal microflora during in vitro fermentation.

This study was to investigate the potential prebiotic function of exopolysaccharide (EPS) from a medicinal fungus and the relationship to the molecular properties by in vitro human fecal fermentation. The EPS from Cordyceps sinensis Cs-HK1 mycelial fermentation was processed into three fractions with different monosaccharide contents, a higher molecular weight (MW) and a lower MW attained by two-step ethanol precipitation, and an intermediate MW by ultrasound-degradation of EPS. All the EPS fractions were well utilized during 24-48 h of fecal fermentation, leading to significant increases in the short chain fatty acid (SCFA) production. The consumption rate and production level of SCFAs varied slightly with the different EPS fractions. The EPS also influenced the composition and diversity of the fecal microflora, increasing the relative abundance of Firmicutes but suppressing that of Proteobacteria, which may be a beneficial effect for human health. Overall the results have shown that the Cs-HK1 EPS has significant prebiotic activity which is dependent on its molecular properties.

Protective effects of exopolysaccharide of a medicinal fungus on probiotic bacteria during cold storage and simulated gastrointestinal conditions.

The efficacy of probiotic health products depends on the capability of the constituent probiotic bacteria to survive through long period of cold storage and the gastrointestinal tract. This study was to evaluate the protective effects of a high-molecular weight (MW) exopolysaccharide (EPS) from a medicinal fungus Cs-HK1 on three different bifidobacteria. The EPS had a total dietary fiber content about 70% (w/w), which was close to its total carbohydrate content. It was resistant to artificial gastric acid (pH 2) with no more than 4% (w/w) hydrolysis in 6 h. EPS at 5 g/L significantly increased the survival rate of the probiotic bacteria during cold storage (4 °C) and in simulated gastric acid, reducing the death rate of different bacterial strains by 50% to 70%. The protective effect of EPS was weaker when the concentration was decreased to 3 g/L or when the MW of EPS was reduced by partial degradation with power ultrasound. EPS also showed significantly protective effect on the all bacterial strains in bile juice. The results have demonstrated the potential value of Cs-HK1 EPS as a novel prebiotic fiber for the formulation of synbiotic products with probiotic bacteria.

Protection of Bifidobacterial cells against antibiotics by a high molecular weight exopolysaccharide of a medicinal fungus Cs-HK1 through physical interactions.

This study was to assess the protective effect of exopolysaccharide (EPS) produced by a medicinal fungus Cordyceps sinensis Cs-HK1 on Bifidobacteria against antibiotic inhibition. The high-molecular weight EPS fractions showed significant protective effect on all five bifidobacterial strains against four common antibiotics, leading to a dramatic increase in the minimal inhibitory and minimal bactericidal concentrations. The protective effect of EPS on the bacteria was probably attributed to the formation of a viscous layer around the bacterial cell resisting the access by the antibiotics. The EPS layer surrounding the bacteria cell also promoted the aggregation of bacteria and formation of biofilm as observed by microscopy. EPS also enhanced the bifidobacterial adhesion to Caco-2 cell monolayer. In general, the protective effect as well as biofilm formation due to EPS was significantly correlated with the molecular weight of EPS fractions.

Cosmetic and Skincare Benefits of Cultivated Mycelia from the Chinese Caterpillar Mushroom, Ophiocordyceps sinensis (Ascomycetes).

Mushrooms are potential sources of novel natural cosmeceutical ingredients. This study was conducted to evaluate the cosmetic (skincare) benefits of the valuable medicinal species Ophiocordyceps sinensis (=Cordyceps sinensis). The mycelial extracts of 2 O. sinensis strains, Cs-HK1 and Cs-4, prepared sequentially with ethyl acetate, ethanol, and hot water were tested with in vitro assays for tyrosinase-, collagenase-, and elastase-inhibitory activity. The ethyl acetate extracts of both fungal strains showed potent antityrosinase and antielastase activity, with low half-maximal inhibitory concentrations (0.14-0.47 mg/mL) comparable to those of the respective reference compounds (arbutin and epigallocatechin gallate). All mycelial extracts exhibited moderate or significant anticollagenase activity; most extracts showed a significant photoprotective effect with a sun protection factor up to 25. The results from this study show the potential use of O. sinensis as a source of cosmetic ingredients for skincare applications.

Bifidogenic effects of Cordyceps sinensis fungal exopolysaccharide and konjac glucomannan after ultrasound and acid degradation.

The bifidogenic effects of exopolysaccharide (EPS) of a medicinal fungus (Cordyceps sinensis) and a well-known food polysaccharide konjac glucomannan (KGM) with different molecular weight (MW) ranges were evaluated through in vitro experiments in liquid cultures of Bifidobacteria. Native EPS and KGM were partially degraded with power ultrasound (US) to improve the water solubility, and further hydrolysed with trifluoroacetic acid to much lower MW. The acid-hydrolysed fractions (EPS-AH and KGM-AH) supported the growth of all five tested bifidobacterial species, while the US-degraded high MW fractions, EPS-US and KGM-US, could only slightly support the growth of some species. All EPS fractions increased the acetic acid production of most bifidobacterial species. Most remarkably, the high MW EPS-US, EPS-AH and KGM-US fractions significantly enhanced the cell viability with much higher colony forming unit (CFU) counts, suggesting a protective effect of these high MW polysaccharides for the bacterial survival. The results have shown that MW was a significant factor on the bifidogenic properties of partially degraded EPS and KGM.

Protective effects of natural and partially degraded konjac glucomannan on Bifidobacteria against antibiotic damage.

This study was to evaluate the protective effects of a dietary fiber, konjac glucomannan (KGM) from the plant tuber of Amorphohallus konjac on Bifidobacteria against antibiotic damage. KGM (∼8.8×108Da) was partially degraded with high-intensity ultrasound to KGM-US (∼1.8×106Da) and then hydrolyzed with trifluoroacetic acid (TFA) to KGM-AH (1369Da). KGM-US (at 5g/l) showed the most significant protective effect on most bifidobacterial strains against penicillin and streptomycin inhibition, increasing the minimal inhibitory and bactericidal concentration (MIC and MBC) dramatically, and KGM also showed significant effects on enhancing the MBC of enrofloxacin, penicillin, tetracycline and streptomycin. In addition, the adsorbance ability and biofilm formation effects of KGM and degraded KGM products may be partially responsible for the protective effects. The results suggested that natural KGM and ultrasound treated KGM have protective effects for the human gut probiotic bacteria against the damage caused by specific antibiotics.

Formation and Physiochemical Properties of Silver Nanoparticles with Various Exopolysaccharides of a Medicinal Fungus in Aqueous Solution.

Natural polysaccharides are the most widely used biopolymers for green synthesis of eco-friendly silver nanoparticles (AgNPs). In a previous study, a high molecular weight (MW) fraction of exopolysaccharides (EPS) produced by a medicinal fungus Cs-HK1 has been shown useful for green and facile synthesis of AgNPs in water. This study was to further evaluate the effects of molecular properties of EPS on the formation, stability and properties of AgNPs with different EPS fractions at various pH conditions. Three EPS fractions (P0.5, P2.0 and P5.0: MW high to low and protein content low to high) were reacted with silver nitrate at various pH 3.0-8.0 in water. The most favorable pH range was 5.5-8.0 for the formation and stable dispersion of AgNPs. At a given pH, the maximum amount of AgNPs was produced with P5.0, and the minimum with P0.5. The shape, size and physiochemical properties of AgNPs were strongly affected by the molecular characteristics of EPS (MW and conformation). The results may be helpful for understanding the factors and mechanisms for formation of stable AgNPs with natural polysaccharides and the interactions between AgNPs and the polysaccharide hydrocolloids in water.