Sephadex G-100 Chromatography Research Articles

Structural Characterization of Polygonatum Cyrtonema Polysaccharide and Its Immunomodulatory Effects on Macrophages.

A neutral Polygonatum cyrtonema polysaccharide (NPCP) was isolated and purified from Polygonatum cyrtonema by various chromatographic techniques, including DEAE-52 and Sephadex-G100 chromatography. The structure of NPCP was characterized by HPLC, HPGPC, GC-MS, FT-IR, NMR, and SEM. Results showed that NPCP is composed of glucose (55.4%) and galactose (44.6%) with a molecular weight of 3.2 kDa, and the sugar chain of NPCP was →1)-α-D-Glc-(4→1)-β-D-Gal-(3→. In vitro bioactivity experiments demonstrated that NPCP significantly enhanced macrophages proliferation and phagocytosis while inhibiting the M1 polarization induced by LPS as well as the M2 polarization induced by IL-4 and IL-13 in macrophages. Additionally, NPCP suppressed the secretion of IL-6 and TNF-α in both M1 and M2 cells but promoted the secretion of IL-10. These results suggest that NPCP could serve as an immunomodulatory agent with potential applications in anti-inflammatory therapy.

Ultrasound assisted hot water extraction of polysaccharides from Taraxacum mongolicum: Optimization, purification, structure characterization, and antioxidant activity.

Ultrasound assisted hot water extraction (UAHWE) was applied to extraction of polysaccharides from Taraxacum mongolicum with hot water as extract solvent. Experimental factors in UAHWE process were optimized by response surface methodology. The optimal extraction parameters to achieve the highest Taraxacum mongolicum polysaccharides (TMPs) yield (12.08±0.14)% by UAHWE were obtained under the ultrasound power of 200W, extraction temperature of 62°C, solid-to-liquid ratio of 1:20g/mL, and extraction time of 40min, and then the crude TMPs were further purified by DEAE-52 and Sephadex G-100 chromatography to obtain a homogenous polysaccharide fraction (TMPs-1-SG). Subsequently, the structure of TMPs-1-SG was characterized by UV-vis, Fourier transform infrared spectroscopy (FT-IR), high performance gel permeation chromatography (HPGPC), high performance liquid chromatography (HPLC), scanning electron microscope (SEM), transmission electron microscopy (TEM), and Congo red test. The results display that TMPs-1-SG with an average molecular weight of 5.49 × 104Da was comprised of mannose (Man), galactose (Gal), xylose (Xyl), and arabinose (Ara) with a molar ratio of 39.85:52.61:27.14:6.30. Moreover, TMPs-1-SG did not contain a triple helix structure. Furthermore, TMPs-1-SG and TEM presented a sheet-like, rod-shaped, and irregular structure. Finally, the antioxidant activity of TMPs-1-SG was evaluated by in vitro experiment. The IC50 values of scavenging DPPH and OH radicals for TMPs-1-SG achieved 0.71mg/mL and 0.75mg/mL, respectively. The findings can provide an effective method for extracting polysaccharides from natural resources.

Purification, kinetic characterization of thermostable multicopper oxidase from the oyster mushroom and its versatility for greener agro-pulp bio bleaching in the paper industry.

Production of a thermostable laccase from Pleurotus florida was reported for the first time, both in submerged and solid-state fermentation using agro-industrial residues. This enzyme was purified using ammonium sulphate precipitation (60-90%), Sephadex G-100 and DEAE column ion exchange chromatography, respectively. The laccase was purified to 21.49 fold with an apparent molecular weight of 66 kDa and had an optimal pH of 5 with temperature stability at 60°C. Metal ions such as Cu2+ (91.26 µmole/mL/min), Mg2+ (68.15 µmole/mL/min), and Fe2+ (1.73 µmole/mL/min) enhanced the laccase activity, but Fe2+ (1.73µmole/mL/min) inhibited the enzyme activity. The purified laccase had Km and Vmax of 16.68 mM and 26.73 µmole/mL/min for guaiacol as a substrate. The isolated enzyme was characterized by FT-IR which revealed bands at 3655.0 cm-1, 2894.7 cm-1, and 1151.7 cm-1 corresponding to primary amines, C-H stretch, and amide -III, respectively. The enzymatic bio bleaching of paddy straw pulp was found to be most effective which resulted in a lowering of kappa number and yellowness by 19.47% & 17.84% whereas an increase in brightness and whiteness by 41.92%. & -19.61%. Thus, this might be stated that the crude laccase from P. florida can be exploited to reduce the toxic waste load for managing environmental pollution and helps in enhancing the yield and quality of the paper.

Isolation, Purification, Fractionation, and Hepatoprotective Activity of Polygonatum Polysaccharides.

In this study, three homogeneous fractions, PSP-N-b-1, PSP-N-b-2, and PSP-N-c-1, were obtained from an aqueous extract of Polygonatum using DEAE cellulose column chromatography, CL-6B agarose gel chromatography, and Sephadex G100 chromatography. Their monosaccharide compositions and molecular weights were analyzed. The results revealed that PSP-N-b-1, PSP-N-b-2, and PSP-N-c-1 are primarily composed of six monosaccharides: Man (mannose), GlcA (glucuronic acid), Rha (rhamnose), GalA (galacturonic acid), Glc (glucose), and Ara (arabinose), with molecular weights of 6.3 KDa, 5.78 KDa, and 3.45 KDa, respectively. Furthermore, we observed that Polygonatum polysaccharides exhibited protective effects against CCL4-induced liver damage in HepG2 cells in vitro, operating through both anti-oxidant and anti-inflammatory mechanisms. Our research findings suggest that Polygonatum polysaccharides may emerge as a promising option in the development of hepatoprotective drugs or functional foods with anti-inflammatory and antioxidant properties.

Extraction, purification, and investigation of the antibacterial potential of lysozyme isolated from the latex of Calotropis procera

Calotropis procera produces a latex used in traditional medicine because of its components which are found to be biologically active. The latex of C. procera has many benefits such as pain reduction, antimicrobial attitude and other precious attributes. Latex lysozyme (LL) is a particular protein contained in C. procera plant having a promising antibacterial property. The process applied for extraction implicated ammonium sulfate precipitation, water extraction, ion exchange and gel filtration column chromatography evidencing 14.4 kDa for LL as a molecular mass. LL after purification showed 397.18 U mg−1 as specific activity (10.30 purification fold), and with 37.9% as a yield. In addition, the chemical and physical analyses showed that LL had its best realization when the pH was 5.5 at 50 °C. Moreover when metal ions such as Ca2+ and Mg2+ were incorporated the relative activity of the enzyme resulted ampliflied. When a purification was conducted with CM-Cellulose and Sephadex G-100 chromatography, LL revealed an antibacterial property versus both Escherichia coli (MICs 14 µg ml−1) and Bacillus cereus (MICs 13 µg ml−1). HR-TEM analysis displayed an antimicrobial potential of LL after its purification; that analysis showed the deterioration of the cell wall (external membrane of the cell) of both E. coli and B. cereus. Therefore, the LL obtained from C. procera maintains ability as an antibacterial enzyme applicable in various domains such as biotechnological and pharmaceutical industries.Graphical abstract

Purification and partial characterization of a thermostable peroxidase isoenzyme from Bambara groundnut (Vigna subterranea L. Verdc) seedlings

Thermostable isoperoxidases, especially peroxidase A6, have already been found in seedlings of Vigna sp, indicating their potential for biotechnological applications. In this study, peroxidase A6 was purified from the roots of Bambara groundnut (Vigna subterranea L. Verdc) seedlings by a combination of gel filtration on Sephadex G-100, heat treatment, and ion exchange chromatography on CM cellulose and DEAE cellulose. The 41-kDa enzyme shows high catalytic efficiency in the oxidation of O-dianisidine, ABTS, TMB, DAB and OPD at acidic pH optimum and the reduction of H2O2. It has an optimum temperature of activity of 60 °C and a calculated activation energy of 221.5 KJ/mol for its thermal inactivation at pH 8. Mg2+ inhibits the enzyme. Ca2+ strongly increases its thermal stability, while Mn2+ and Zn2+ decrease it. The enzyme is inhibited by sodium azide at concentrations above 1 µM, with an IC50 value of about 10 µM. This inhibition, in addition to the RZ value of 2.4 (A403nm/A280nm), confirms the presence of a haem group in the active site, which is common for class III plant peroxidases. Due to its unique catalytic and thermal properties, peroxidase A6 has the potential to be a valuable tool for various biotechnological applications, especially for enzyme immunoassays and clinical diagnosis.

Antibacterial and anticancer activities of (free and immobilized) elastase produced by Klebsiella pneumoniae

Elastase is a type of protease that degrades explicitly elastin. The elastase produced by Klebsiella pneumoniae isolates was purified by three steps: ammonium sulfate precipitation, ion-exchange chromatography, and Sephadex G-150 chromatography. The optimal condition for elastase production showed high specific activity with starch (3.8 U/mg protein) and casein as a nitrogen source with a specific activity reaching (3.3 U/mg protein). The maximum elastase production was obtained when the pH value was (7.5) with specific activity (4.4 U/mg protein). Elastase (free and immobilized on TiO2- NPs) was used in application as antibacterial and anticancer, and results showed high antibacterial activity against pathogenic isolates, especially Lactobacillus acidophilus and Pseudomonas aeruginosa were affected by immobilized elastase. Free and immobilized elastase have anticancer activity against lung cancer using the A549 cell line, and immobilized elastase had the potent cytotoxic effect on A549 cells with IC50 142.8 µg/ ml compared with IC50 of normal cell line HdFn on 655.0 µg /ml. Key Words: Klebsiella pneumoniae, Elastase, Immobilization, TiO2-Nps- Antibacterial, Anticancer.

A novel heteropolysaccharide isolated from custard apple pulp and its immunomodulatory activity in mouse macrophages and dendritic cells

In this study, a novel heteropolysaccharide (ASPA80-1) with an average molecular weight of 5.48 × 104 Da was isolated and structurally elucidated from custard apple pulp (Annona squamosa) through DEAE-cellulose, Sephadex G-100 and Sephacryl S-300 HR chromatography and spectral analysis. ASPA80-1 is a water-soluble polysaccharide and it is a polymer consisting of predominant amounts of (1 → 3)-linked-L-arabinose (Ara) residues, small amounts of (1 → 6)-linked-D-galactose (Gal), (1 → 3,5)-linked-L-arabinose (Ara) residues and terminal linked-L-arabinose (Ara) residues, trace amount of (1 → 4)-linked-D-glucose (Glc) residues and (1 → 2)-linked-L-rhamnose (Rham) residues. ASPA80-1 showed significant effect on antigen-presenting cells (APCs) activation. On the one hand, ASPA80-1 activated RAW264.7 macrophage cells by inducing morphology change, enhancing phagocytic ability, increasing nitric oxide (NO) secretion and promoting expression of major histocompatibility complex class II (MHC II) and cluster of differentiation 86 (CD 86). On the other hand, ASPA80-1 promoted the maturation of dendritic cells (DCs) by inducing longer dendrites, decreasing phagocytic ability and increasing MHC II and CD86 expression. Furthermore, mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B (NF-κB) signaling pathways were activated after the intervention of ASPA80-1 on RAW264.7 cells or DCs. Thus, the novel heteropolysaccharide ASPA80-1 has the potential to be used as an immunoenhancing component in functional foods.

Production, optimization, and purification of alkaline thermotolerant protease from newly isolated Phalaris minor seeds

The present work aims to purify and perform a preliminary analysis on a thermostable serine alkaline protease from a recently identified P. minor. The enzyme was purified 2.7-fold with a 12.4 % recovery using Sephadex G-100 chromatography, DEAE-cellulose, and ammonium sulphate precipitation. The isolated enzyme has a specific activity of 473 U/mg. The purified protease had a molecular mass of 29 kDa, and just one band was seen, which matched the band obtained using SDS-PAGE. High thermostability was demonstrated by the enzymes, which had half-lives of 31.79 and 6.0 min (a 5.3-fold improvement), enthalpies of denaturation (ΔH°) of 119.53 and 119.35 KJ mol−1, entropies of denaturation (ΔS°) of 32.96 and 41.11 J/mol·K, and free energies of denaturation (ΔG°) of 108.87 and 105.58 KJ mol−1 for the protease enzyme. Studies on the folding and stability of alkaline proteases are important since their use in biotechnology requires that they operate in settings of extreme pH and temperature. According to the kinetic and thermodynamic properties, the protease produced by P. minor is superior to that produced by other sources and previously described plants, and it might find utility in a variety of industrial fields.

Preparation of nano-selenium from chestnut polysaccharide and characterization of its antioxidant activity.

Chestnut is widely cultivated and has high nutritional value due to its richness in polysaccharides. In order to improve the antioxidant activity of chestnut polysaccharide, chestnut polysaccharide (CP) was extracted by ultrasonic-assisted water extraction and alcohol precipitation and purified by cellulose DEAE-52 exchange and Sephadex G-100 chromatography in this study. CP isolates were characterized by I2-KI reaction, three-strand helical structure analysis, infrared spectrum analysis, and nuclear magnetic resonance detection. The results showed that CP is a pyrylan sugar with triple helical structure and connected by α-glycosidic bonds, with sugar residues 1,4-α-D-Glcp, 1,6-α-D-Galp, 1,5-α-L-Araf, 1,4-α-L-Rhap, and 1,4-β-D-Glcp in the CP backbone. After purification, the branching structure, rod, and spherical structure were significantly increased, with reduced lamellar structure. The in vitro scavenging rates of CP at 10 mg·mL-1 against DPPH, hydroxyl radicals, and ABTS were 88.95, 41.38, and 48.16%, respectively. The DPPH free radical scavenging rate of purified polysaccharide fraction CP-1a was slightly enhanced, and the other rates showed a small decrease. Selenized chestnut polysaccharide (CP-Se) was prepared using nano-selenium method. The selenization method was optimized and stable Se-CP was obtained. When the concentration was 5 mg·mL-1, Se-CP had significantly higher scavenging abilities 89.81 ± 2.33, 58.50 ± 1.60, and 40.66 ± 1.91% for DPPH, hydroxyl radical, and ABTS radicals, respectively, than those of CP. The results of this study provide insight into the effects purification and selenization of chestnut polysaccharide on antioxidant activity, and also provide a theoretical basis for the development of chestnut polysaccharide for use in functional foods or health products.

Purification and biochemical characterization of two laccase isoenzymes isolated from Trichoderma harzianum S7113 and its application for bisphenol A degradation

Two laccase isoenzymes (LacA and LacB) were isolated from a novel Trichoderma harzianum S7113 isolate employing ammonium sulfate precipitation, Sephadex G100, and DEAE Sepharose ion exchange chromatography. The molecular weights of the purified LacA and LacB laccases were estimated to be 63 and 48 kDa, respectively. The two isoenzymes had their optimum activities at the same temperature (50 °C), but at slightly different pH values (pH 3.0 for LacA and pH 2.5 for LacB). LacA and LacB had the same thermal stability at 40 °C and pH stability at pH 9.0. The two isoenzymes also showed a high level of specific activity toward ABTS, where the Km values of LacA and LacB were 0.100 and 0.065 mM, whereas their Vmax values were 0.603 and 0.182 µmol min−1, respectively. LacA and LacB catalytic activity was stimulated by Mg2+, Zn2+, K+, and Ni2+, whereas it was inhibited by Hg2+ and Pb2+, β-mercaptoethanol, EDTA, and SDS, and completely inhibited by sodium azide. Our findings indicate that purified laccase has a promising capacity for bisphenol A (BPA) bioremediation across a broad pH range. This finding opens up new opportunities for the commercialization of this technique in a variety of biotechnology-based applications, particularly for removing endocrine chemicals from the environment.

A novel cyclic dipeptide from Lactiplantibacillus plantarum MC39 inhibits proliferation of multidrug-resistant Klebsiella pneumoniae W8 and Enterobacter hormaechei U25

The present study pays special attention to lactic acid bacteria products, cyclic dipeptides (CDPs), as an effective prevention strategy that can be used to eradicate the threats posed by multidrug-resistant (MDR) Enterobacteriaceae. Herein, Lactiplantibacillus plantarum MC39, a strain isolated from an artisanal Egyptian cheese, was a producer of a novel CDP that could strongly inhibit, in vitro, MDR Klebsiella pneumoniae W8 and Enterobacter hormaechei U25, for the first time. Following partial purification with 80% ammonium sulphate precipitation and Sephadex G-100 chromatography, this biomolecule was identified as the cyclic dipeptide cyclo(Ala–Glu) using IR, HNMR, and LC–MS. Cyclo(Ala–Glu) was thermostable up to 121 °C, insensitive to pepsin and α-chymotrypsin, tolerant to organic solvents, active at acidic pH, and stable for 45 days of storage. Thus, it shows clear evidence to be usable as a bio-preservative in dairy products and as a potential therapeutic agent, controlling MDR K. pneumoniae and E. hormaechei.

Purification, structural characterization and immunological activity of Sibiraea laexigata (L.) Maxim polysaccharide.

Sibiraea laexigata (L.) Maxim (SLM) has been used as an herbal tea for treating stomach discomfort and indigestion for a long time in china. Polysaccharides have been identified as one of the major bioactive compounds in the SLM. In the present paper, ultrasonic-assisted enzymatic extraction (UAEE) method was employed in polysaccharides extraction derived from SLM using polyethylene glycol (PEG) as extraction solvent, two SLM polysaccharides (SLMPs) fractions (SLMPs-1-1 and SLMPs-2-1) were purified by DEAE Cellulose-52 and Sephadex G-100 chromatography in sequence. Then, the preliminarily structure of the two factions were characterized by chemical composition analysis, molecular weight measurement, UVS, HPLC-PMP, FT-IR, nuclear magnetic resonance (NMR) spectra analysis and SEM. The results showed that SLMPs-1-1 and SLMPs-2-1 with different molecular weights of 1.03 and 1.02 kDa, mainly composed of glucose (46.76 and 46.79%), respectively. The results of structural characterization from FT-IR, 1H NMR, and SEM revealed that SLMPs-1-1 and SLMPs-2-1 contained the typical pyranoid polysaccharide with α-glycosidic bond and β-glycosidic bond. Furthermore, it was found that SLMPs-1-1 could increase the levels of tumor necrosis factor-α (TNF-α) and interleukin-2 (IL-2), and alleviated the immune organs tissue damage of cyclophosphamide (Cy)-treated mice. RT-qPCR and Western-Blot analysis showed that SLMPs-1-1 could significantly up-regulated the levels of NF-κB, TLR4, which revealed that SLMPs-1-1 could participate in immunosuppressive protection of Cy-treated mice. These findings suggested that the potential of SLMPs-1-1 as an alternative immunostimulator could be used in food and pharmaceutical industries.

Microwave assisted extraction, characterization of a polysaccharide from Salvia miltiorrhiza Bunge and its antioxidant effects via ferroptosis-mediated activation of the Nrf2/HO-1 pathway

A microwave-assisted extraction procedure for the crude Salvia miltiorrhiza polysaccharides (SMPs) obtained from Salvia miltiorrhiza Bunge was optimized. Four independent variables were studied: microwave power, extraction time, solvent-to-solid ratio, and concentration of ethanol, with optimal settings of 1200 W, 12 min, 38, and 86 %, respectively. The SMPs were successively purified by DEAE Sepharose Fast Flow and Sephadex G-100 chromatography to produce a novel polysaccharide termed SMP1. The SMP1 was composed of glucose, galactose, and fructose in a molar ratio of 1:1.67:1.12 with an average molecular weight of 6087 Da. Pharmacological studies showed that SMP1 protected from OGD/R-induced ferroptosis and lipid peroxidation by activating Nrf2/HO-1 pathway in PC12 cells. Our research systematically indicated that polysaccharide could inhibit ferroptosis to alleviate oxidative stress injury, which laid the foundation for the future clinical application of Salvia miltiorrhiza polysaccharide.

Purification of wild type and recombinant xylanases and evaluation of their arabinoxylan hydrolysis ability

Arabinoxylan hydrolyzing enzymes are very rich and diversified groups, among these, endo-1,4-β-xylanase is the most important group which catalyzes the cleavage of xylan main skeleton to release arabinoxylan oligosaccharides such as D-xylose, xylobiose, L-arabinose, xylotetraose, xylopentose. This group of enzyme was efficiently produced from natural microorganism including bacteria, yeast and fungi and recombinant strains. In this study, we have purified wild-type and recombinant xylanase to compare the difference in hydrolysis products of these two enzymes. Using sephadex G-100 and DEAE-sephadex column chromatography, we have purified xylanase from wild-type Aspergillus niger DSM1957 with molecular weight of 25 kDa, specific activity of 3240 IU/mL, the purity increases 1.68 folds compare to crude extract, and a recovery rate of 36%. The recombinant enzyme from Pichia pastoris GS115/pPXlnA was purified by ProbondTM affinity chromatography column, with a molecular weight of 36 kDa; the purity increased 3.2 folds than the crude extract, and with the recovery efficiency of 21.1%. The activity of commercial wild-type xylanase was better than that of our recombinant and wild-type enzymes. Our wild-type xylanase could hydrolyze arabinoxylan into L-arabinose in 50 mM CH3COONa pH 5.0. Our results showed that xylanases purified from different sources were stable and highly specific to xylan. Xylanase has potential application in the production of high quality biotechnological products.

Modeling, optimization, purification, and characterization of polysaccharides from Lilium lancifolium Thunb

Ultrasound assisted aqueous two-phase extraction of polysaccharides from Lilium lancifolium Thunb was modeled by response surface methodology (RSM) and artificial neural network (ANN), and optimized using genetic algorithm coupled with ANN (GA-ANN). Statistical analysis showed that the models obtained by RSM and ANN could accurately predict the Lilium lancifolium Thunb polysaccharides (LLPs) yield. However, ANN prediction was more accurate than RSM. The optimum extraction parameters to achieve the highest LLPs yield (15.17 ± 0.21)% was obtained at the ultrasound power of 250 W, extraction temperature of 63 °C, liquid-to-solid ratio of 21 mL/g, and extraction time of 32 min. Subsequently, the crude LLPs were further purified via DEAE-52 and Sephadex G-100 chromatography to obtain a homogenous fraction (LLPs-2-SG, 421.41 kDa) that contained mannose, glucose, and galactose in a molar ratio of 10.52:23.06:7.19. The structure of LLPs-2-SG was characterized with UV–vis, FT–IR, AFM, and SEM. The findings provide a critical method for the extraction, separation and purification of polysaccharides from natural resources.

Isolation, Structural Characterization and Macrophage Activation Activity of an Acidic Polysaccharide from Raspberry Pulp.

The discovery of safe and effective plant polysaccharides with immunomodulatory effects has become a research hotspot. Raspberry is an essential commercial fruit and is widely distributed, cultivated, and consumed worldwide. In the present study, a homogeneous acidic polysaccharide (RPP-2a), with a weight-average molecular weight (Mw) of 55582 Da, was isolated from the pulp of raspberries through DEAE-Sepharose Fast Flow and Sephadex G-200 chromatography. RPP-2a consisted of rhamnose, arabinose, galactose, glucose, xylose, galacturonic acid and glucuronic acid, with a molar ratio of 15.4:9.6:7.6:3.2:9.1:54.3:0.8. The results of Fourier transform infrared spectroscopy (FT-IR), gas chromatography-mass spectrometer (GC-MS), 1D-, and 2D-nuclear magnetic resonance (NMR) analyses suggested that the backbone of RPP-2a was primarily composed of →2)-α-L-Rhap-(1→, →2,4)-α-L-Rhap-(1→, →4)-α-D-GalAp-(1→, and →3,4)-α-D-Glcp-(1→ sugar moieties, with side chains of α-L-Araf-(1→, α-L-Arap-(1→, and β-D-Galp-(1→3)-β-D-Galp-(1→ residues linked to the O-4 band of rhamnose and O-3 band of glucose residues. Furthermore, RPP-2a exhibited significant macrophage activation activity by increasing the production of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β), and the expression of inducible nitric oxide synthase (iNOS) and cytokines at the transcriptional level in RAW264.7 cells. Overall, the results indicate that RPP-2a can be utilized as a potential natural immune-enhancing agent.

Ultrasound assisted aqueous two-phase extraction of polysaccharides from Cornus officinalis fruit: Modeling, optimization, purification, and characterization

Ultrasound assisted aqueous two-phase extraction of polysaccharides from Cornus officinalis fruit was modeled by response surface methodology (RSM) and artificial neural network (ANN), and optimized using genetic algorithm coupled with ANN (GA-ANN). Statistical analysis showed that the models obtained by RSM and ANN could accurately predict the Cornus officinalis polysaccharides (COPs) yield. However, ANN prediction was more accurate than RSM. The optimum extraction parameters to achieve the highest COPs yield (7.85 ± 0.09)% was obtained at the ultrasound power of 350 W, extraction temperature of 51 ℃, liquid-to-solid ratio of 17 mL/g, and extraction time of 38 min. Subsequently, the crude COPs were further purified via DEAE-52 and Sephadex G-100 chromatography to obtain a homogenous fraction (COPs-4-SG, 33.64 kDa) that contained galacturonic acid, arabinose, mannose, glucose, and galactose in a molar ratio of 34.82:14.19:6.75:13.48:12.26. The structure of COPs-4-SG was also characterized with UV–vis, fourier-transform infrared spectroscopy (FT–IR), atomic force microscopy (AFM), scanning electron microscopy (SEM), Congo-red test, and circular dichroism (CD). The findings provide a feasible way for the extraction, purification, and optimization of polysaccharides from plant resources

Polysaccharides from Spores of Cordyceps cicadae Protect against Cyclophosphamide-Induced Immunosuppression and Oxidative Stress in Mice

This study investigated the purification, preliminary structure and in vivo immunomodulatory activities of polysaccharides from the spores of Cordyceps cicadae (CCSP). The crude CCSP was purified by diethylaminoethyl (DEAE)-cellulose and Sephadex G-100 chromatography, affording CCSP-1, CCSP-2 and CCSP-3 with molecular weights of 1.79 × 106, 5.74 × 104 and 7.93 × 103 Da, respectively. CCSP-2 consisted of mannose and glucose, while CCSP-1 and CCSP-3 are composed of three and four monosaccharides with different molar ratios, respectively. CCSP-2 exhibited its ameliorative effects in cyclophosphamide-induced immunosuppressed mice through significantly increasing spleen and thymus indices, enhancing macrophage phagocytic activity, stimulating splenocyte proliferation, improving natural killer (NK) cytotoxicity, improving bone marrow suppression, regulating the secretion of cytokines and immunoglobulins, and modulating antioxidant enzyme system. These results indicate that CCSP-2 might be exploited as a promising natural immunomodulator.

Characterization and diabetic wound healing benefits of protein-polysaccharide complexes isolated from an animal ethno-medicine Periplaneta americana L.

Periplaneta americana L. (PA), a type of animal medicine, has been widely used for wound healing in clinical settings. In order to further investigate the bioactive wound healing substances in PA, crude PA protein-polysaccharide complexes were further purified by cellulose DE-52 and Sephadex G100 chromatography in succession. Among these isolated fractions, two fractions eluted by 0.3 M and 0.5 M NaCl with the higher yield, respectively named PaPPc2 and PaPPc3 respectively, were chosen for the wound healing experiments. Mediated by HPGPC, amino acid and monosaccharide composition analysis, circular dichroism spectrum, glycosylation type, FT-IR, and 1H NMR analysis, the characterization of PaPPc2 and PaPPc3 was implemented. And then, the benefits of PaPPcs to promote cell proliferation, migration, and tube formation of HUVECs were determined in vitro, indicated these fractions would facilitate angiogenesis. Finally, as proof of concept, PaPPc2 and PaPPc3 were employed to accelerate the acute wounds of diabetic mice, involving in increase blood vessels and the amounts of angiogenesis-related cytokines (α-SMA, VEGF, and CD31). In short, this study provides an experimental basis to demonstrate the protein-polysaccharide complexes of Periplaneta americana L. as its wound healing bioactive substances.