High-performance Gel Filtration Chromatography Research Articles

Structural characteristics, immune-activating mechanisms in vitro, and immunomodulatory effects in vivo of the exopolysaccharide EPS53 from Streptococcus thermophilus XJ53

Our previous investigations have successfully identified the repeating structural units of EPS53, an exopolysaccharide derived from Streptococcus thermophilus XJ53 fermented milk, and substantiated its potential immunomodulatory properties. The present study further elucidated the structural characteristics of EPS53 and investigated the underlying mechanisms governing its in vitro immunoreactivity as well as its in vivo immunoreactivity. The results obtained from multi-detector high performance gel filtration chromatography revealed that EPS53 adopted a rigid rod conformation in aqueous solution, with the weight-average molecular weight of 1464 kDa, the number-average molecular weight of 694 kDa, and the polydispersity index of 2.11. Congo red experiment confirmed the absence of a triple helix conformation. Scanning electron microscopy showed that EPS53 displayed a three-dimensional fibrous structure covered with flakes. The in vitro findings indicated that EPS53 enhanced phagocytosis ability, reactive oxygen species (ROS) production, and cytokine levels of macrophages via the TLR4-mediated NF-κB/MAPK signaling pathways as confirmed by immunofluorescence staining experiments, inhibition blocking experiments, and Western blot assay. Additionally, the in vivo experiments demonstrated that EPS53 significantly increased macrophage and neutrophil number while enhancing NO and ROS levels in zebrafish larvae; thus, providing further evidence for the immunomodulatory efficacy of EPS53.

Characterization and in vitro prebiotic activity analysis of succinoglycan produced by Rhizobium radiobacter

In this study, a novel succinoglycan (COGs-3) was produced and purified from Rhizobium radiobacter ATCC 13333, and its structural characteristics, physical properties, and in vitro prebiotic activity were investigated. The COGs-3 production was 2.761 g·L−1 after single-factor and response surface optimizations. The molecular weight and molecular formula of COGs-3 were 1584 Da and C59H92O49. The structure of COGs-3 consisted of β-Glcp-(1→3)-β-Glcp-(1→3)-β-Glcp-(1→6)-β-Glcp-(1→6)-β-Glcp-(1→4)-β-Glcp-(1→4)-β-Glcp-(1→3)-α-Galp modified with two succinate and one pyruvate groups based on ultraviolet–visible, Fourier transform infrared spectroscopy, high-performance gel filtration chromatography, elemental, monosaccharide composition, matrix-assisted laser desorption/ionization time of flight tandem mass spectrometry, methylation, and nuclear magnetic resonance analyses, which was the type of succinoglycan. In addition, the thermal gravimetric and differential scanning calorimetry analyses showed that 40.23 % of COGs-3 maintained physical integrity when the temperature reached 600 ℃ and the melting point was 171.42 °C, which confirmed that COGs-3 had good thermal stability. The effects of COGs-3 on healthy human gut microbiota in vitro were also studied. COGs-3 was not digested by digestive juice and significantly increased the relative abundances of Bifidobacterium and Lactococcus and decreased the abundance of Megamonas and klebsiella. In addition, COGs-3 considerably promoted short-chain fatty acids biosynthesis. This work presents the first report on the in vitro prebiotic properties of succinoglycan, thereby providing new ideas for the application and development of succinoglycan.

Pharmacodynamic structure of deer antler base protein and its mammary gland hyperplasia inhibition mechanism by mediating Raf-1/MEK/ERK signaling pathway activation.

Mammary gland hyperplasia (MGH) is a common mammary disease whose main pathogenesis is the disruption of estradiol (E2) and progesterone (P) secretion, thereby causing overproliferation of mammary epithelial cells and mammary gland tissue hyperplasia. Deer antler base is a traditional Chinese medicine that has been used for many years to treat MGH. However, its pharmacological mechanism and pharmacodynamic material basis are unclear. In this study, we for the first time used the graded salting method to classify deer antler base protein (CNCP) as CNCP-A, CNCP-B, and CNCP-C and explored the pharmacological mechanism of the anti-MGH properties of CNCP. We found that CNCP could regulate the hormonal levels of E2, P, and follicle stimulating hormone (FSH) and improve the histopathological condition. The potential mechanism might be related to the recombinant C-Raf proto oncogene serine/threonine protein kinase/mitogen-activated protein/extracellular regulated protein kinase (Raf-1/MEK/ERK) signaling pathway. By upregulating the protein expression of the follicle stimulating hormone receptor (FSHR), cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) inhibited the activation of the downstream Raf-1/MEK/ERK signaling pathway, which in turn inhibited the proliferation of mammary epithelial cells. We analyzed the physicochemical properties of CNCP-A, CNCP-B, and CNCP-C and obtained CNCP-C-I by column chromatographic purification of the best pharmacophore protein CNCP. Using high-performance liquid gel filtration chromatography (HPGFC), we determined the molecular weight of CNCP-C-I and identified it by high-performance liquid tandem mass spectrometry (LC-MS/MS) to obtain the first match for a high confidence protein KRT1. This study provides a theoretical basis for the development of effective traditional Chinese medicines with low toxicity levels for the prevention and treatment of mammary gland diseases.

Emulsification characteristics of soy hull polysaccharides obtained by membrane separation

Membrane separation technology was used to separate and purify the microwave-assisted oxalic acid extraction of soy hull polysaccharides (MOSP) in order to obtain samples of different molecular weights. The emulsification characteristics of the MOSP were investigated including protein adsorption, polysaccharide adsorption, interfacial tension, emulsion index, and particle size; optical microscopy and Phenom electron microscopy were used to elucidate the emulsion structures. In addition, Fourier transform infrared spectroscopy (FT-IR), atomic force microscopy (AFM), and high-performance gel-filtration chromatography (HPGFC) were used to study the differences in the components and structures of MOSP in different molecular weights. The molecular weight had several important effects on the emulsifying properties of MOSP. The adsorption capacities of the emulsion droplets containing low molecular weight MOSP (L-MOSP), middle molecular weight MOSP (M-MOSP), and high molecular weight MOSP (H-MOSP) were relatively low, and those of H-MOSP were slightly higher than those of L-MOSP. With extended storage time, the particle sizes of the emulsions rich in L-MOSP, M-MOSP, and H-MOSP increased. L-MOSP, M-MOSP, and H-MOSP were mainly composed of furans. The conformation of the molecular chain was spherical. The emulsions formed with H-MOSP were the most stable.

Analysis of the polymerized impurities in cefmetazole sodium based on novel separation principle by liquid chromatography tandem ion trap/time-of-flight mass spectrometry

To effectively control the polymerized impurities in cefmetazole sodium, novel high performance gel filtration chromatography (HPSEC) with TSK-gel G2000SWxl column and RP-HPLC method with C18 column were used in replace of classical gel filtration chromatography with Sephadex G-10 gel. By studying the chromatographic behavior of polymerized impurities in both chromatographic systems with different chromatographic separation principles, the polymerized impurities in cefmetazole sodium were separated and detected effectively. The two-dimensional liquid chromatography tandem ion trap/time-of-flight mass spectrometry (2D LC-IT-TOF MS) was applied to characterize the structures of polymerized impurities eluted from HPSEC method, and liquid chromatography tandem ion trap/time-of-flight mass spectrometry was applied to characterize the structures of polymerized impurities and other unknown impurities eluted from RP-HPLC method. The structures of fourteen unknown impurities in cefmetazole sodium were deduced based on the MS n data, nine of which were polymerized impurities. The corresponding relationship between impurities in the HPSEC method and RP-HPLC method was established, and the specificity of the two methods was evaluated. The RP-HPLC method for analysis of the polymerized impurities has higher column efficiency and specificity than the HPSEC method. The RP-HPLC method is suitable for quality control of the polymerized impurities in cefmetazole sodium. The forming mechanisms of degradation impurities in cefmetazole sodium were also studied.

High-Efficiency Production of Auricularia polytricha Polysaccharides Through Yellow Slurry Water Fermentation and Its Structure and Antioxidant Properties.

Yellow slurry water is a kind of nutrient-rich wastewater of tofu. Firstly, the medium of yellow slurry was optimized. Then, APP40, APP60, and APP80 were obtained by sedimentation with different concentration of ethanol (40, 60, and 80%). The physicochemical properties and primary structures of the three polysaccharides were studied by high performance anion exchange chromatography (HPAEC), high performance gel filtration chromatography (HPGFC), scanning electron microscope (SEM), atomic force microscope (AFM), and Fourier transform infrared (FT-IR) spectrometer. Finally, the effects of three polysaccharides on antioxidation activity were studied. According to the experimental optimization the results, the biomass and the production of Auricularia polytricha Polysaccharides (APPS) reached the peak, and they were 13.5 ± .655 and 9.42 ± .253 g/L (p < .05). The SEM and the AFM showed that the height of APP80 gradually increased from 31.1 to 46.7 nm and from APP40 to APP80. The particle size of APP80 increased, the pores decrease or even disappear, and the molecules begin to aggregate. The FT-IR spectrum analysis showed that the three polysaccharides possessed key functional groups. The carbohydrate content of APP40, APP60, and APP80 was 20.2, 34.25, and 31.73%. The molecular weights of APP40, APP60, and APP80 are 9.462 × 104, 8.742 × 104, and 8.091 × 104 Da, respectively. The three polysaccharides were composed of rhamnose, galactose, glucose, mannose, and xylose but with different molar ratio. APP80 showed strong reducing ability and scavenging activity of 2,2–diphenyl–1–picrylhydrazyl (DPPH) and hydroxyl radicals through antioxidant activities evaluated in vitro. This study introduces a way for the effective use of yellow slurry water.

Multiple-fingerprint analysis of Poria cocos polysaccharide by HPLC combined with chemometrics methods

In this study, the multiple fingerprints, which were integrated with HPGFC-ELSD (high performance gel filtration chromatography - evaporative light scattering detector) fingerprint, PMP-HPLC-DAD (1-phenyl-3-methyl-5-pyrazolone-high performance liquid chromatography - diode array detector) fingerprint of complete acid hydrolysates and HILIC-HPLC-ELSD (hydrophilic interaction - high performance liquid chromatography - evaporative light scattering detector) fingerprint of enzyme hydrolysates, were established to evaluate the quality of polysaccharides from Poria cocos (PCPs). The similarity evaluation showed that 16 batches of PCPs from different origins had high similarity in structural characteristics based on the multiple fingerprints. The chromatographic data of multiple fingerprints of PCPs were fused, processed and analyzed by chemometric methods including HCA (hierarchical cluster analysis), PCA (principal component analysis) and PLS-DA (partial least squares discriminant analysis). The 16 batches of PCPs were divided into 3 categories in PCA, indicating a certain relationship between the structural characteristics and the origins. PLS-DA analysis indicated that Man, Glc, Gal, Fuc, the components with m/z of 2.22 × 104∼1.53 × 104 Da and 3.46 × 103∼2.69 × 103 Da, oligosaccharides with DPs of 6 and 7, respectively, could be regarded as potential chemical markers for the classification of PCPs from different origins. According to the multiple fingerprints and chemometric analysis, the two commercial samples were proved to be adulterants.

Isolation of peanut protein aggregates using aqueous extraction processing combined with membrane separation

SummaryA two‐stage microfiltration and ultrafiltration (MF/UF) for peanut protein recovery and water recycling from aqueous extraction processing (AEP) were investigated. Peanut protein aggregates and alkaline water were obtained using flat and wound MF/UF and alkaline water recycled in AEP. With this approach, most of the wastewater could be recycled to produce reusable water to solve wastewater problems in aqueous oil extraction processing from oilseeds. The result showed that a little oil and protein were lost (1.21% and 4.35%, respectively) during the recycling of permeate in AEP, which was still acceptable. Peanut protein aggregations (PPAs) were characterised by analysis with Fourier‐transform infrared spectroscopy (FTIR), sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS‐PAGE), high‐performance gel filtration chromatography (HPGFC) and binding force analysis. Solubility analysis suggested that PPA had molecular structures with more hydrophobic groups on the surface than in the peanut protein isolate (PPI). In vitro, PPA was digested by both pepsin and trypsin more than PPI. FTIR profiles demonstrated there were more β‐sheet and less α‐helix in PPA, which means much more aggregation. In addition, binding force analysis showed hydrophobic interaction was the major force that restricted dissolution of PPA and disulfide bonds were of second importance, which provided a possible physical treatment opportunity for improving the solubility of PPA.

Immunomodulatory and Antioxidant Activities of a Polysaccharide from Ligustrum vicaryi L. Fruit.

Ligustrum vicaryi L. is a hybrid of Ligustrum ovalifolium Hassk. var. aureo-marginatum and Ligustrum vulgale L., belonging to the Oleaceae family. It is often used as an ornamental shrub due to its golden leaves. However, its medical value is yet to be discovered. Recently, plant polysaccharides have attracted comprehensive attention owing to their biological properties, including immunomodulatory and antioxidant activities. This study aimed to extract, purify, and characterize the polysaccharide from the Ligustrum vicaryi L. fruit and investigate its immunomodulatory and antioxidant activities. The Ligustrum vicaryi L. fruit polysaccharide (LVFP) was obtained by ultrasonic extraction, ethanol precipitation, macroporous resin separation, and dialysis bag purification. The physicochemical properties of the LVFP were elucidated using Fourier-transform infrared spectrometry, high-performance ion chromatography, and high-performance gel filtration chromatography. The results indicated that the LVFP consisted of rhamnose, arabinose, galactose, and glucose in a ratio of 1.79 : 7.55 : 4.58 : 1.54, and its molecular weight was 88,949 Da. The immunomodulatory and antioxidant activities of the LVFP were investigated using a cyclophosphamide- (Cy-) induced immunosuppressed mouse model. The results demonstrated that the LVFP significantly increased spleen and thymus indexes, enhanced the phagocytic function of neutrophils, activated B and T lymphocytes, and upregulated serum levels of IL-10 and TNF-α. Moreover, we observed that the LVFP relieved Cy-induced liver damage by increasing superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) levels. These results suggested that the LVFP has the immunomodulatory and antioxidant activities, therefore laying a foundation for the application of the LVFP in the pharmaceutical and functional food industries.

Use ofKluyveromyces marxianusprefermented wheat bran as a source of enzyme mixture to improve dough performance and bread biochemical properties

AbstractBackground and objectivesIn this study,Kluyveromyces marxianuswith ability to produce β‐glucosidase was used to preferment wheat bran (WB) and release a mixture of natural enzymes for improving dough performance and bread quality. Five different enzyme activities were analyzed. The effect of the prefermented WB with enzyme mixture on stickiness and microstructure of dough, ferulic acid release in bread, and molecular weight distribution of arabinoxylan were evaluated using Texture Analyzer, scanning electron microscopy (SEM), HPLC, and high‐performance gel filtration chromatography (HPGFC).FindingsKluyveromyces marxianusexhibited abundant extracellular β‐glucosidase activity (up to 6.98 U/g). The main enzymes released in prefermented bran were endoglucanase and exoglucanase, β‐glucosidase, xylanase, and ferulate esterases. These enzymes were active during bread making (only prefermented bran added as improver or combined with xylanase) and affected solubility and molecular weight distribution of dietary fiber, which resulted in positive improvement of dough performance, release of phenolic acid, and overall quality of the breads. The changes observed were attributed to the extracellular enzymes fromK. marxianusand endogenous enzymes activated during prefermentation, thus enhancing the WB functionality in bread.ConclusionsWheat bran bioprocessing using high enzyme producing yeasts increases functional application of wheat bran in baked products.Significance and noveltyThis study added new knowledge for developing a natural enzyme‐based improver for bran‐enriched baked products in the food industry.

Isolation, Characterization, and Pharmaceutical Applications of an Exopolysaccharide from Aerococcus Uriaeequi.

Many marine bacteria secrete exopolysaccharides (EPSs), which are made up of a substantial component of the macro-molecules surrounding cells. Recently, the wide demand for EPSs for food, cosmetics, pharmaceutical and other applications has led to great interest in them. In this study, an EPS produced by marine bacteria Aerococcus uriaeequi HZ strains (EPS-A) was isolated and purified to examine its structure and biological function. The molecular weight of EPS-A analyzed by high-performance liquid gel filtration chromatography (HPGFC) is found to have a number average of 2.22 × 105 and weight average of 2.84 × 105, respectively. High-performance liquid chromatography (HPLC) and Fourier-transform–infrared (FT–IR) analysis indicate that EPS-A was a polysaccharide composed of glucose and a little mannose. In addition, the flocculating rate of sewage of EPS-A was 79.90%. The hygroscopicity studies showed that hygroscopicity of EPS-A was higher than chitosan but lower than that of sodium hyaluronate. The moisture retention of EPS-A showed similar retention activity to both chitosan and sodium hyaluronate. EPS-A also can scavenge free radicals including both OH• free radical and O2•− free radical and the activity to O2•− free radical is similar to vitamin C. Safety assessment on mice indicated that the EPS-A is safe for external use and oral administration. EPS-A has great potential for applications in medicine due to its characteristics mentioned above.

Structure Characterization of Honey-Processed Astragalus Polysaccharides and Its Anti-Inflammatory Activity In Vitro.

Honey-processed Astragalus is a dosage form of Radix Astragalus mixed with honey by a traditional Chinese medicine processing method which strengthens the tonic effect. Astragalus polysaccharide (APS), perform its immunomodulatory effects by relying on the tonic effect of Radix Astragalus, therefore, the improved pharmacological activity of honey-processed Astragalus polysaccharide (HAPS) might be due to structural changes during processing. The molecular weights of HAPS and APS were 1,695,788 Da, 2,047,756 Da, respectively, as determined by high performance gel filtration chromatography combined with evaporative light scattering detection (HPGFC-ELSD). The monosaccharide composition was determined by ultra-performance liquid chromatogram quadrupole time-of-flight mass spectrometry (UPLC/ESI-Q-TOF-MS) after pre-column derivatization with 1-phenyl-3-methyl-5-pyrazolone (PMP). The results showed that the essential components were mannose, glucose, xylose, arabinose, glucuronic acid and rhamnose, is molar ratios of 0.06:28.34:0.58:0.24:0.33:0.21 and 0.27:12.83:1.63:0.71:1.04:0.56, respectively. FT-IR and NMR analysis of HAPS results showed the presence of uronic acid and acetyl groups. The anti-inflammatory activities of HAPS were more effective than those of APS according to the NO contents and the expression of IFN-γ, IL-1β, IL-22 and TNF-α in lipopolysaccharide (LPS)-induced RAW264.7 cells. This findings suggest that the anti-inflammatory and bioactivity improvement might be associated with molecular structure changes, bearing on the potential immunomodulatory action.

Relative molecular mass and monosaccharide composition of polysaccharide in Dendrobium officinale flowers

In order to provide new information on polysaccharide in Dendrobium officinale flowers,the monosaccharide composition and relative molecular mass distribution of 11 families of flowers were investigated and analyzed by high performance gel filtration chromatography (HPGFC) and pre-column derivatization ultra performance liquid chromatography (UPLC) in this study. Then cluster analysis was carried out for the monosaccharide peak areas by utilizing SPSS 19.0 software. The results showed that the polysaccharides of all the above 11 hybrid families of D. officinale flower were separated into three fractions (DOP-1, DOP-2 and DOP-3) with the average relative molecular mass of 5.53×105, 3.49×105 and 2.12×105. The polysaccharides in 11 different families were mainly composed of glucose, mannose, galactose, galacturonic acid and arabinose; mannose had the highest proportion among them, with mannose/glucose ratio of 0.302-3.335. Additionally, the relative contents of various monosaccharides in different families varied. 11 families of D. officinale flower could be classified into four categories according to their monosaccharide components and relative contents. In this study, the relative molecular mass distribution and monosaccharide composition of polysaccharides in D. officinale flowers were defined, which can provide foundations for its resource utilization．.

Structural Characterization and Antifatigue Effect In Vivo of Maca (Lepidium meyenii Walp) Polysaccharide.

Maca (Lepidium meyenii Walp) polysaccharides (MP) with purity of 99.2% were obtained to investigate their structural characteristics and antifatigue effect in vivo. The physicochemical properties of MP were analyzed through high-performance gel filtration chromatography, IR, monosaccharide composition, methylation, GC-MS, and NMR analyses. The antifatigue effect of MP was evaluated by using a mouse weight-loaded swimming model. MP is an acidic heteropolysaccharide with an average molecular weight (Mw ) of 793.5 kDa. It is composed of D-GalA: D-Glc: L-Ara: D-Man: D-Gal: L-Rha = 35.07:29.98:16.98:13.01:4.21:0.75 (mol, %). The findings revealed that MP contained β-1,3-Galp(A), β-1,3-Glcp, and α-1, 3-Manp linked alternatingly to form a backbone (5:4:1). MP (above mid-dosage 50 mg/kg bw/d) could effectively elongate swimming durations and accelerate average swimming speeds (within the 1st 5 min) of mice (P < 0.05) and improve the serous biochemical parameters of mice. Compared with the control model, high-dosage (100 mg/kg bw/d) MP treatment could significantly enhance glutathione peroxidase and creatine kinase activities (P < 0.05) and decreased lactate dehydrogenase activity (P < 0.01). High-dosage MP could significantly reduce the levels of blood urea nitrogen, lactic acid, and malondialdehyde (P < 0.05). MP is an acidic polysaccharide with a high D-GalA content, which could be responsible for the antifatigue effect of maca.

Discussion and Analysis of &lt;em&gt;Flammulina velutipes Polysaccharides&lt;/em&gt; Compositions, Molecular Weight and Monosaccharide Composition

The aim of the present work was to explore the composition, molecular weight and monosaccharide composition of ,Flammulina velutipes polysaccharides by DEAE Cellulose-52, High Performance Gel Filtration Chromatography (HPGFC) and HPLC with PMP derivative with the FVP isolated and purified from Flammulina velutipes. The results showed that Flammulina velutipes polysaccharides were composed of three kinds of polysaccharides; the molecular weight of FVP was composed of 4191338, 372779, 19002; the molecular weight of 372779, 19002, one of the biggest average molecular weight polysaccharides is neutral polysaccharide, the rest of the two kinds of polysaccharide composition of less average molecular weight is acidic polysaccharides and the proportion of two kind of acidic polysaccharides significantly greater than neutral polysaccharide, 44.31 and 37.76% respectively, only 17.93% of neutral polysaccharide. FVP was composed of Glc, Man, Gal, Xyl, Fuc; the highest content of Glc, followed by Gal and Man, Xyl and Fuc content is relatively low. The molar ratio was relatively 13.05:2.75:3.16:1.48:1.00. In conclusion, FVP contained the more Glc, the less Fuc and a small amount of galacturonic acid and glucuronic acid, which showed FVP may contain acidic polysaccharides.

Inulin Molecular Detection Methods of High-Performance Gel Filtration Chromatography and Changes in Inulin Molecular Weight During Storage

Inulin Molecular Detection Methods of High-Performance Gel Filtration Chromatography and Changes in Inulin Molecular Weight During Storage

Isolation, purification, characterization and immunostimulatory activity of polysaccharides derived from American ginseng

In this study, crude American ginseng polysaccharide (AGPS) was extracted with hot water and preliminarily purified by using resin S-8 and Polyamide columns. Then, it was further purified and separated by DEAE-Sepharose CL-6B and Sepharose CL-6B chromatography, respectively. Five main fractions were obtained, named WPS-1, WPS-2, SPS-1, SPS-2 and SPS-3. Their homogeneities and structural characteristics were elucidated based on UV–vis spectroscopy, High Performance Gel Filtration Chromatography (HPGFC), Gas Chromatography (GC), Scanning Electron Microscopy (SEM), Infrared Spectrum (IR), and NMR Spectroscopy methods. Furthermore, the immunostimulatory effects of these fractions upon splenic lymphocyte proliferation, macrophage phagocytosis and nitric oxide (NO) production, were investigated in vitro. The results indicated that their stimulations could be ordered as SPS-3>SPS-1>CPS (crude polysaccharides)>WPS-1>WPS-2>SPS-2. Among them, SPS-3 showed more potent immunomodulatory activity and could be explored as a potential immunopotentiating agent for use in functional food or medicine.

Separation and purification of low-molecular-weight chondroitin sulfates and their anti-oxidant properties

Low-molecular-weight chondroitin sulfate was obtained by degradation of chondroitin sulfate using hyaluronidase. Then, separated with sephadex G25, DEAE-52, and finally purified with AKATA superpeptide separation system and fluorescence-assisted carbohydrate electrophoresis. The main compo-nents detected by high performance gel-filtration chromatography were disaccharide, tetrasaccharide, hexasaccharide with molecular weight of 521, 1024 and 1527 Da, respectively. The anti-oxidant activity of these three oligosaccharides in vitro showed that the reducing power (maximum value at 10 mg/mL) and superoxide anion radical scavenging abilities were increased (maximum value at 4 mg/mL) with an increased in their concentration. There were no significant differences of the anti-oxidant properties between those three oligosaccharides.

Characterization and In vitro Antioxidant Activity of a Polysaccharide from C ordyceps sobolifera

The characteristics and antioxidant activities of Cordyceps sobolifera polysaccharide (CSP) were investigated. Results from high-performance gel filtration chromatography showed that the average molecular weight of CSP was 1.53 × 104 kDa, while this polysaccharide was mainly composed of mannose, glucose and galactose in the molar ratio of 1.7:8.9:1 as determined by high-performance liquid chromatography after pre-column derivatization with 1-phenyl-3-methyl-5-pyrazolone. Fourier-transform infrared spectrum of CSP indicated the typical functional groups of polysaccharide. Antioxidant activity results showed that CSP had reducing power (OD700 was 0.87 at 1.5 mg/mL), and it also exhibited scavenging activities on hydroxyl (·OH scavenging rate was 47.6% at 5 mg/mL) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals (scavenging rate was 55.6% at 1 mg/mL) in a concentration-dependent manner. These results indicate that CSP has good in vitro antioxidant activity and can potentially be used for functional foods. Practical Applications Cordyceps sobolifera is a pricy traditional Chinese herb and it has been used in China for a long time. Polysaccharide extracted from C. sobolifera was characterized, and the antioxidant activities of this polysaccharide were investigated. The result may lay the foundation for the development of antioxidant functional foods from C. sobolifera.

Finding of polysaccharide-peptide complexes in Cordyceps militaris and evaluation of its acetylcholinesterase inhibition activity

Acetylcholinesterase (AChE) inhibition enhances learning and cognitive ability for treatment of Alzheimer's disease. Polysaccharide–peptide complexes were identified in Cordyceps militaris (CPSPs) and characterized for their AChE inhibitory properties. Three polymers (CPSP-F1, -F2, and -F3) were extracted and separated by ultrasound-assisted extraction and diethylaminoethanol (DEAE)–Sepharose CL-6B column chromatography. Polysaccharide–peptide complexes were identified by DEAE–Sepharose CL-6B column chromatography and high-performance gel-filtration chromatography, Fourier transform infrared spectra, amino sugar composition analysis, and β-elimination reaction to identify polysaccharide–peptide bond categories. Separation of CPSP can increase AChE inhibitory activity from the crude polysaccharide of C. militaris. CPSP-F1 and CPSP-F2 exhibited half maximal inhibitory concentrations of 32.2 ± 0.2 mg/mL and 5.3 ± 0.0 mg/mL. Thus, we identified polysaccharide–peptide complexes from C. militaris and suggest CPSP has great potential in AChE inhibition bioassay.