Sephadex G-100 Gel Column Research Articles

Structure identification and activity evaluation of polysaccharide from Codonopsis pilosula (C. pilosula nannf. Var. modesta (nannf.) L. T. Shen)

Codonopsis pilosula is a medicinal plant with properties related to food, and the antioxidant and hypoglycemic components of its natural polysaccharides, as well as their related mechanisms, have not been fully understood. This study examined the physicochemical properties and structure of C. pilosula polysaccharide (WCP) using an orthogonal extraction method involving hot water-alcohol precipitation, high-resolution spectroscopy, and microscopic imaging technology. The antioxidant and hypoglycemic activities of the polysaccharides were assessed. Six fractions (WCP1, WCP2, WCP3, WCP4, WCP5, and WCP6) were identified through purification using a DEAE-cellulose column and Sephadex G-100 gel column. Mannose, glucose, and arabinose were identified as the primary monosaccharide components of WCPs through Gas Chromatography (GC). The polysaccharide exhibited a crystal structure composed of irregularly entangled pyranose particles of WCPs as revealed by X-ray diffraction (XRD), scanning electron microscopy (SEM), and atomic force microscopy (AFM). The polysaccharide decomposed into smoother triple-helix fibrous flocs as indicated by Congo red analysis and circular dichroism analysis (CD). The antioxidant capacity of the polysaccharides against various free radicals was evaluated, with WCP4 demonstrating a significant reducing capacity. Moreover, WCP3 and WCP5 displayed notable inhibition rates of α-amylase and α-glucosidase, respectively. Molecular docking studies indicated the interaction of WCP5 with two enzymes, forming multiple hydrogen bonds through specific amino acids. In the gromacs simulation, both composite systems maintained a steady state in terms of root mean square deviation (RMSD), root mean square fluctuation (RMSF), and radius of gyration, providing strong support for their potential as a reference for the developing novel therapeutic approaches to diabetes.

Studies of the Immunomodulatory Activity of Polysaccharides from the Stem of Cynomorium songaricum Based on Intestinal Microbial Analysis.

Polysaccharides are the main effective components of Cynomorium songaricum's stem that perform biological activities and have positive impacts on immune enhancement. In this study, the polysaccharide CSP-III of Cynomorium songaricum's stem was isolated using a DEAE-52 cellulose column through Sephadex G-100 gel column chromatography. Upon analysis, the monosaccharide composition of CSP-III included Mannose (Man), Glucuronic acid (GlcA), Galacturonic acid (GalA), Rhamnose (Rha), Glucose (Glc), Galactose (Gal), and Arabinose (Ara), at a molar ratio of 0.01:0.11:0.03:0.57:0.02:0.32:1. The molecular weight of CSP-III was 4018234 Da. Meanwhile, the capacity of CSP-III, at various concentrations, to stimulate the proliferation of mouse spleen lymphocytes in vitro was compared, and the influence of CSP-III on cell proliferation was examined using RAW264.7 mouse mononuclear macrophages as a model. The influence of CSP-III on the expression of important phosphorylating proteins in the MAPK signaling pathway was initially analyzed by Western blotting. In RAW264.7 cells, CSP-III promoted the phosphorylation of JNK proteins, which thus activated the MAPK signaling cascade and exerted immunomodulatory effects. Moreover, according to in vivo studies using cyclophosphamide (CTX)-induced immunosuppression mouse models, CSP-III improved the CTX-induced histopathological damage, promoted T and B lymphocyte proliferation, upregulated CD4+ and CD8+ T-lymphocyte counts in the spleen, increased the serum levels of IgG and IgM, and activated three essential proteins of the MAPK signaling pathway. As revealed by analysis of intestinal flora, CSP-III improved the immune function by maintaining the homeostasis of the bacterial flora by boosting the relative abundances of some beneficial bacterial groups, such as Bacteroidetes, Desmodium, and Actinomyces, and reducing the relative abundance of Aspergillus phylum. Through in vitro and in vivo experiments, our present study demonstrates that polysaccharides from the stem of Cynomorium songaricum possess strong immunoregulatory effects. Findings in this work provide theoretical support for the potential application of Cynomorium songaricum in the field of health food.

Preparation, structural characterization and in vitro activity of ginger polysaccharide

AbstractA polysaccharide was extracted from the fresh ginger by water extraction and alcohol precipitation. A homogeneous polysaccharide was obtained by Sevage deproteinization, dialysis, DEAE-52 ion exchange chromatography and Sephadex G-100 gel column chromatography. The molecular weight of the ginger polysaccharide was determined by high performance gel permeation chromatography. The monosaccharide composition, scanning electron microscope and other physicochemical parameters of the ginger polysaccharide were further determined. The spectral properties of the ginger polysaccharide were investigated by ultraviolet and infrared spectroscopy. On this basis, methylation analysis, 1D and 2D NMR analysis were used to investigate its possible chemical structure. The results showed that the ginger polysaccharide group was divided into neutral polysaccharide components with high homogeneity, Mw = 35.52 kDa, Mn = 216.56 kDa and the sugar content was 93.5%. The ginger polysaccharide is mainly composed of Fuc, Ara, Rha, Gal, Glc, Xyl, Man, Gal-UA and Glc-UA, the amount of substance ratio was 1.01:1.96:9.54:14.82:64.20:3.84:2.07:1.41:1.16. To study the structure–activity relationship of ginger polysaccharide, acetylated ginger polysaccharide and phosphate ginger polysaccharide were prepared by chemical methods in this study. The results of antioxidant activity in vitro showed that the ginger polysaccharide and its derivatives had good biological activity in freeing ABTS + radical scavenging, freeing hydroxyl radical scavenging, freeing DPPH radical scavenging and reducing ability. Interestingly, the results showed that the ginger polysaccharide had a certain inhibitory effect on the growth of E. coli in vitro. In general, this study provides a certain experimental basis and reference for the further research on the activity of ginger polysaccharide and its derivatives. Graphical Abstract

Characterization of a novel polysaccharide from red ginseng and its ameliorative effect on oxidative stress injury in myocardial ischemia

BackgroundRed ginseng (RG) was widely used as traditional Chinese medicine (TCM) or dietary supplement. However, few researches had been reported on the red ginseng polysaccharide (RGP).MethodsIn this study, a novel heteropolysaccharide named RGP1-1 was fractionated sequentially by DEAE-52 column and Sephadex G-100 gel column. The primary structure of RGP1-1, including glycosyl linkages, molecular weight, monosaccharide composition, morphology and physicochemical property were conducted by nuclear magnetic resonance (NMR), gas chromatography-mass spectrometer (GC–MS), atomic force microscope (AFM), scanning electron microscope (SEM), differential scanning calorimetry-thermogravimetric analysis (DSC-TG) and so on. The effect of RGP1-1 in preventing and treating myocardial ischemia was evaluated by an animal model isoprenaline (ISO) induced mice.ResultsRGP1-1, with a homogeneous molecular weight of 5655 Da, was composed of Glc and Gal in the ratio of 94.26:4.92. The methylation and NMR analysis indicated the backbone was composed of → 1)-Glcp-(4 → and → 1)-Galp-(4 →, branched partially at O-4 with α-D-Glcp-(1 → residue. Morphology and physicochemical property analysis revealed a triple-helical conformation, flaky and irregular spherical structure with molecule aggregations and stable thermal properties of RGP1-1. And it contained 6.82 mV zeta potential, 117.4 nm partical size and polymerization phenomenon. Furthermore, RGP1-1 possessed strong antioxidant activity in vitro and in vivo, RGP1-1 could decrease cardiomyocyte apoptosis and myocardium fibrosis of mice in histopathology and it could decrease significantly the serum levels of cardiac troponin (cTnI), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), malondialdehyde (MDA). Western blot analysis showed that RGP1-1 can increase the expression of main protein Nuclear factor E2-related factor 2(Nrf2), NAD(P)H:quinone oxidoreductase 1 (NQO1), heme oxygenase-1(HO-1) and kelch-like ECH-associated protein1(keap1) in oxidative stress injure progress, and therefore regulate the pathway of Nrf2/HO-1.ConclusionThe above findings indicated that RGP1-1 had an improving effect on ISO-induced myocardial ischemia injury in mice, as novel natural antioxidant and heart-protecting drugs.

Isolation, Structural Analysis and Anti-Inflammatory Activity of a Polysaccharide from Ilex cornuta Fruits.

In the present study, a polysaccharide from Ilex cornuta fruits (LCFP-3) was obtained by hot water extraction, Diethyaminoethyl cellulose-52 (DEAE-52) chromatography column and Sephadex G-100 gel column purification. Its structural characteristics were further explored using high performance anion exchange chromatography (HPAEC), gas chromatography and mass spectrometry (GC/MS), scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy and nuclear magnetic resonance (NMR) spectroscopy. Monosaccharide composition analysis revealed LCFP-3 contained mainly Galactose (31.92 %), Arabinose (25.87 %) and Galacturonic acid (23.35 %) while small percentage of Rhamnose, Glucose, Mannose and Xylose. Chemical composition analysis showed that the total sugar content of LCFP-3 was 90.31 % and the protein content was 0.246 %. Gel permeation chromatography (GPC) analysis showed that its average molecular weight was 41.199 kDa. Structural analysis showed that LCFP-3 may be composed of residues, T-α-Arap, T-α-Rhap, 1,3-α-Arap, 1,4-α-Arap, T-β-Galp, 1,4-α-GalpA(OMe), 1,4-β-Glcp, 1,3-β-Galp, 1,3,6-β-Manp, 1,6-β-Galp, 1,3,4-β-GalpA, 1,4,6-β-Manp, 1,3,6-β-Glcp, 1,2,3,4-α-Xylp. The anti-inflammatory activity of LCFP-3 was evaluated using lipopolysaccharide (LPS)-induced RAW246.7 macrophages. The results showed that 1-200 μg/mL LCFP-3 could dose-dependently protect against LPS-induced toxicity and 1 μg/mL LCFP-3 could significantly inhibit LPS-induced NO production. Therefore, LCFP-3 exerted an anti-inflammatory activity and has great potential as a functional ingredient.

Preparation of Active Peptides from Camellia vietnamensis and Their Metabolic Effects in Alcohol-Induced Liver Injury Cells

Liver damage seriously affects human health. Over 35% of cases of acute liver damage are caused by alcohol damage. Thus, finding drugs that can inhibit and effectively treat this disease is necessary. This article mainly focuses on the effect of the metabolome physical activity of active peptides in Camellia vietnamensis active peptide (CMAP) and improving liver protection. DEAE Sepharose FF ion-exchange column chromatography was used in separating and purifying crude peptides from Camellia vietnamensis Two components, A1 and A2, were obtained, and the most active A1 was selected. Sephadex G-100 gel column chromatography was used in A1 separation and purification. Three components, Al-1, Al-2, and Al-3, were obtained. Through antioxidant activity in vitro as an index of inspection, the relatively active component A1-2 was removed. Reverse-phase high-performance liquid chromatography showed that the purity of component A1-2 was 93.45%. The extracted CMAPs acted on alcoholic liver injury cells. Metabolomics studies revealed that the up-regulated metabolites were ribothymidine and xanthine; the down-regulated metabolites were hydroxyphenyllactic acid, creatinine, stearoylcarnitine, and inosine. This study provides an effective theoretical support for subsequent research.

Purification, biochemical characterization, and molecular cloning of cellulase from Bacillus licheniformis strain Z9 isolated from soil

BackgroundCellulose is the most prevalent biomass and renewable energy source in nature. The hydrolysis of cellulosic biomass to glucose units is essential for the economic exploitation of this natural resource. Cellulase enzyme, which is largely generated by bacteria and fungus, is commonly used to degrade cellulose. Cellulases are used in a variety of industries, including bioethanol manufacturing, textiles, detergents, drugs, food, and paper. As part of our quest to find an efficient biocatalyst for the hydrolysis of cellulosic biomass, we describe the amplification, cloning, and sequencing of cellulase (cel9z) from Bacillus licheniformis strain Z9, as well as the characterization of the resulting enzyme. ResultsCellulase was partially purified from B. licheniformis strain Z9 using (NH4)2SO4 precipitation and Sephadex G-100 gel column chromatography with 356.5 U/mg specific activity, 2.1-purification fold, and 3.07 % yield. The nucleotide sequence of the cellulase gene was deposited to the GenBank, B. licheniformis strain Z9 cellulase (cel9z) gene, under accession number MK814929. This corresponds to 1453 nucleotides gene and encodes for a protein composed of 484 amino acids. Comparison of deduced amino acids sequence to other related cellulases showed that the enzyme cel9z can be classified as a glycoside hydrolase family 9. SDS-PAGE analysis of the purified enzyme revealed that the molecular mass was 54.5 kDa. The optimal enzyme activity was observed at pH 7.4 and 30 °C. The enzyme was found to be strongly inhibited by Mg2+ and Na+, whereas strongly activated by Fe3+, Cu2+, and Ca2+. ConclusionsB. licheniformis strain Z9 and its cellulase gene can be further utilized for recombinant production of cellulases for industrial application.

Structural elucidation, anti-radical and immunomodulatory activities of polysaccharides from the roots of Glehnia littoralis

This study investigated the structural elucidation, anti-radical and immunomodulatory activities of polysaccharides from the roots of Glehnia littoralis. A crude polysaccharide was extracted from the roots of G. littoralis through the ultrasonic-assisted extraction and further purified by DEAE-52 cellulose and Sephadex G-100 gel column, a major polysaccharide fraction named GLP80-1 was obtained. The chemical properties and structure of GLP and GLP80-1 were characterized by acid hydrolysis, methylation analysis, along with high performance gel permeation chromatography, Fourier transform infrared, nuclear magnetic resonance, scanning electron microscope, thermogravimetric analysis and X-ray diffraction. The molecular weight distributions of GLP were determined as 1.89 × 106 and 1.26 × 104 Da. The monosaccharide composition of GLP was glucose, glucuronic acid, galactose and arabinose with molar ratios of 0.91:0.04:0.03:0.02, respectively. The average molecular weight of GLP80-1 was determined as 1.63 × 104 Da. The structure of GLP80-1 was deduced to be a homogenous glucan, comprised a main chain of (1→4)-linked-α-D-Glcp with a single α-D-Glcp branch substituted at C-6. The results of biological activities in vitro showed that GLP and GLP80-1 exhibited free radical scavenging effects, and displayed promotion for the proliferation of mouse spleen lymphocytes and RAW264.7 cells. The data indicated that GLP and GLP80-1 had the potential to be explored as novel natural antioxidant and immunomodulator for application in functional food.

Protective effects of a polysaccharide from Boletus aereus on S180 tumor-bearing mice and its structural characteristics

A polysaccharide from the aqueous extract of Boletus aereus fruit (BAP) was isolated. The antitumor activities of BAP and/or cyclophosphamide (CTX) were investigated using the model of S180 tumor-bearing mice. Results indicated that BAP could effectively inhibit the growth of S180 solid tumors and protect the immune organs. Hematoxylin and eosin staining, Annexin V-FITC/PI staining, and mitochondrial membrane potential analysis demonstrated that BAP could induce the apoptosis of S180 tumor cells. In combination with CTX, BAP exhibited a significant synergistic antitumor effect on S180 cells. Furthermore, a novel polysaccharide, namely, BAPF, was purified from BAP by using DEAE Cellulose-52 column and Sephadex G-100 gel column. Structural characterization revealed that BAPF was primarily composed of mannose, glucuronic acid, glucose, galactose, arabinose, and fucose at a proportion of 12.98:1:16.8:16.48:1.08:9.1. Its average molecular weight was 1.79 × 106 Da. FTIR and NMR analyses demonstrated that BAPF was a pyranose with α-type and β-type glycosidic residues.

Immunostimulatory effects of the intracellular polysaccharides isolated from liquid culture of Ophiocordyceps sinensis (Ascomycetes) on RAW264.7 cells via the MAPK and PI3K/Akt signaling pathways

Ethnopharmacological relevanceChinese Cordyceps (DongChong XiaCao), a parasitic complex of a fungus Ophiocordyceps sinensis and a caterpillar, is a traditional Chinese medicine. Polysaccharides extracted from O. sinensis have immunomodulatory effects on macrophages. However, the mechanism of polysaccharides on macrophage and the composition of polysaccharides are not known. Aim of studyWe aimed to investigate composition and structure of the intracellular polysaccharides from O. sinensis mycelia (designed as OSP), and evaluate its the immunomodulatory effect on macrophages and its underlying mechanism. Materials and methodsWe performed a liquid-state fermentation of O. sinensis to produce mycelia. The DEAE-Sephadex-A25 cellulose column and Sephadex-G100 gel column chromatography were employed to purify and character the intracellular OSP. Macrophages RAW264.7 cells were employed to evaluate OSP's immunomodulatory activity and the possible mechanism responsible for the activation of macrophages in vitro. ResultsThe average molecular weight of OSP was distributed at 27,972 Da, OSP was composed of xylose, mannose, glucose, and galactose with the ratio of 2.9 : 6.6 : 166 : 2.6, with a trace amount of fucose, arabinose and rhamnose. The phagocytosis of RAW264.7 cells was improved significantly and remarkable changes were observed in the morphology with OSP-treated cells. Real-time quantitative polymerase chain reaction (RT-qPCR) analysis demonstrated that OSP had an ability to regulate the mRNA expression of pro-inflammatory and anti-inflammatory cytokines, and to induce the mRNA expression level of iNOS in a concentration dependent manner in RAW264.7 cells. Western blotting analysis showed that the regulation of NO and cytokines was mediated through mitogen-activated protein kinase (MAPK) and PI3K/Akt signaling pathways. ConclusionThis study demonstrated that OSP was with a capacity to activate macrophage cells RAW264.7 for an improvement of immunomodulation activities, which was through regulation of inflammatory mediators via MAPK and PI3K/Akt signaling pathways.

Glucose- lowering effect of Trametes orientalis polysaccharides in hyperglycemic and hyperlipidemic mice

To explore the glucose-lowering effect of the polysaccharide fractions of Trametes orientalis. The crude polysaccharides of T. orientalis were chromatographed on DEAE cellulose column using H2O and 0.5 mol/L NaCl solution as the eluent and DEAE-water and DEAE-salt with high polysaccharide contents were collected. The two fractions were separated using Sephadex G-100 gel column to obtain 4 polysaccharide fractions TOPW-1, TOPW-2, TOPS-1, and TOPS-2. The anti-oxidation activity of the polysaccharide fractions was investigated with ABTS method. The fractions TOPW-1 and TOPS-1 with consistent UV detection signals were collected and HPGPC was used to determine their relative molecular mass. The monosaccharide composition in homogeneous TOPW-1 was determined by acid hydrolysis combined with HPLC. The in vitro inhibitory activities of TOPW-1 and TOPS-1 against DPP4, adipocyte glucose intake and lipase activity were tested to preliminarily assess their glucose-lowering effect. In a mouse model of hyperglycemic and hyperlipidemic, the glucose-lowering effect of TOPS-1 (0.1, 0.2, and 0.4 g/kg) and its effect on blood lipid metabolism were investigated in comparison with Xiaoke pills (5 pills/kg) and Danshen tablets (0.5 g/kg). TOPW-1 was a homogeneous polysaccharide composed mainly of D-mannose, D-glucose, D-galactose, and D-fucose, with weak antioxidant and hypoglycemic effects. TOPS-1 was not a single polysaccharide and at the concentration of 500 μg/mL showed an high ABTS clearance rate (90.15%). In the mouse model of hyperglycemia and hyperlipidemia, treatment with TOPS-1 (0.2 g/kg) enhanced the activity of lipase and significantly reduced fasting glucose level and serum contents of total cholesterol, triacylglycerol, and low-density lipoprotein cholesterol without causing death in the mice. The glucose-lowering effects of TOPS-1 was not significant at the low (0.1 g/kg) or high (0.4 g/kg) dose, and a high dose tended to increase the mortality of the mice. T. orientalis polysaccharides have anti-oxidation, glucose-lowering and lipid-lowering effects in mice, and their glucose-lowering effect is probably medicated by reducing oxidative stress and ameliorating lipid metabolism disorder.

Extraction, Purification, Characterization, and Antiangiogenic Activity of Acidic Polysaccharide from Buddleja officinalis.

Firstly, optimal parameters of crude polysaccharide from Buddleja officinalis were obtained as follows: ratio of water to raw material of 26 : 1, ultrasonic power of 240 W, ultrasonic time of 45 min, and ultrasonic temperature of 62°C. Secondly, acidic polysaccharide (APBOM) from Buddleja officinalis was successfully acquired with the yield of 9.57% by using DEAE-52 cellulose and Sephadex G-100 gel column chromatography. Then, we found that total polysaccharide content of APBOM was 94.37% with a sulfuric acid group of 1.68%, uronic acid content of 17.41%, and average molecular weight of 165.4 kDa. Finally, APBOM was confirmed to have significant antiangiogenic effects.

Extraction, isolation, immunoregulatory activity, and characterization of Alpiniae oxyphyllae fructus polysaccharides

For further applications of Alpiniae oxyphyllae fructus in modern clinical medicine, Alpiniae oxyphyllae fructus polysaccharide (AOFP) was studied in the present work. The extraction conditions of AOFP were optimized by the response surface method with a Box-Behnken design. The maximum extraction rate of AOFP was 3.18%. An anion-exchange DEAE-52 cellulose column and a Sephadex G-100 gel column were used to isolate the AOFP, and three polysaccharides (AOFP1, AOFP2, AOFP3) were obtained. All three polysaccharides possessed immunoregulatory activity, but the effects of AOFP1 were greater than the other two polysaccharides. AOFP1 significantly stimulated Th1- and Th2-type immune responses and specific immune responses. Meanwhile, the characterization of AOFP1 was studied. AOFP1 was composed of arabinose, galactose, glucose, xylose, mannose, galacturonic acid, and glucuronic acid at a molar ratio of 16.46:12.7:4.9:17.11:4.35:6.52:6 with an average molecular weight of 43.4 kDa. These results suggest that AOFP1 can be developed as a natural immunomodulatory drug.

Characterization, antioxidant, and anticancer activities of a neutral polysaccharide from Duchesnea indica (Andr.) Focke.

A neutral polysaccharide (DIP-1) from Duchesnea indica (Andr.) Focke was obtained by hot water extraction, ethanol precipitation and chromatographic separation (DEAE-52 cellulose anion-exchange column and Sephadex G-100 gel column). The physicochemical properties of DIP-1 were elucidated by gel permeation chromatography, monosaccharide composition, Fourier transform infrared spectrometry, UV-visible spectrophotometry, scanning electron microscope and Congo red test. The results indicated that DIP-1 was consisted of mannose, glucosamine, glucose, galactose and arabinose in a ratio of 1.00:0.42:18.36:14.17:0.81, and its molecular weight was 218.3kDa. Meanwhile, DIP-1 presented a straight hexahedron structure, but no triple-helical conformation. In antioxidant activity tests, DIP-1 exhibited powerful scavenging activities on hydroxyl, DPPH, ABTS radicals and reducing power in a dose-dependent manner. Especially, DIP-1 demonstrated high inhibitory activities against SKOV-3 and Hep-G2 cells in vitro, with IC50 values of 1.42 and 1.23mg/ml, respectively. PRACTICAL APPLICATIONS: D. indica has been used for a long time as a Chinese medicine for therapy of many diseases, including cancer, inflammation, leprosy, fever, bleeding and so on. At present, polysaccharides have attracted comprehensive attention because of a large range of pharmacological and biological properties, including antitumor, antidiabetic, antioxidant and immunomodulatory activity. In the present study, we purified and characterized a neutral polysaccharide from D. indica for the first time. Moreover, the neutral polysaccharide exhibits significant antioxidant and antitumor activities. Therefore, the present study laid a foundation for the high-value application of D. indica polysaccharides in functional food and pharmaceutical industries.

Extraction optimization, characterization, antioxidant and immunomodulatory activities of a novel polysaccharide from the wild mushroom Paxillus involutus

Response surface methodology (RSM) using a Box-Behnken design (BBD) was applied to optimize the extraction of Paxillus involutus polysaccharides. The optimum conditions included an extraction time of 3h, extraction temperature of 79°C and a ratio of liquid to raw material of 43.1mL/g. Under the optimized conditions, the polysaccharides yield was 12.25%. Then, the polysaccharides were purified with DEAE-Cellulose 52 and Sephadex G-100 gel columns, and the fraction denoted as PIP2-1 with a molecular weight of 32kDa was obtained. PIP2-1 was composed of mannose, glucose, galactose, fucose with the mole percentages of 2.8%, 62.2%, 25.4% and 9.6%. The PIP2-1 possessed typical Fourier Transform infrared spectroscopy (FTIR) characterization of polysaccharides. The methylation analysis showed that the PIP2-1 mainly included 1-linked-fuc, 1,3-linked-man, 1-linked-glc, 1,4-linked-glc, 1,6-linked-glc, 1-linked-gal, 1,6-linked-gal, 1,4,6-linked-gal and 1,2,6-gal glycosidic bonds. Furthermore, PIP2-1 showed significant antioxidant activity against hydroxyl radicals (OH), 1,1-diphenyl-2-picrylhydrazyl radicals (DPPH), 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and superoxide radicals. Finally, PIP2-1 significantly enhanced the release of TNF-α and IL-6 in RAW264.7 cells. The results indicated that PIP2-1 could be exploited as a natural antioxidant and immunomodulator for functional food and medical applications.

Purification and characterization of lipase by Bacillus methylotrophicus PS3 under submerged fermentation and its application in detergent industry.

Lipase production bacterial isolate was isolated from soil of service station and identified as Bacillus methylotrophicus PS3 by 16SrRNA with accession number |LN999829.1|. Lipase enzyme was purified by sequential methods of ammonium sulfate precipitation and Sephadex G-100 gel column chromatography. The molecular weight of purified enzyme was 31.40 kDa on SDS-PAGE. This purification procedure resulted in 2.90-fold purification of lipase with a 24.10% final yield. The purified lipase presented maximal hydrolytic activity at a temperature of 55 °C, and pH of 7.0. Lipase activity was stimulated by Triton X-100 and SDS with Mg2+ and Ca2+ metals employ a positive effect and outlast its stable in organic solvent i.e. methanol and ethanol.

Anti-hyperglycemic activity of polysaccharides from calyx of Physalis alkekengi var. franchetii Makino on alloxan-induced mice

The present study was undertaken to investigate the hypoglycemic effect of polysaccharides (PPSC) isolated from calyx of Physalis alkekengi var. franchetii Makino in alloxan-induced diabetic mice. Seven polysaccharide fractions were obtained through a DEAE-52 column and Sephadex G-100 gel column and they were the main composes of PPSC. The PPSC treatment could prevent the loss of body weight in diabetic mice and result in a decrease of fasting blood glucose (FBG) and glycated serum protein (GSP) and an increase of fasting serum insulin in a dose-dependent manner. The histopathological examination of pancreas revealed the ability of PPSC to protect and reverse β-cells from necrosis due destruction of alloxan in diabetic mice. Furthermore, oral PPSC upregulated the expression of PI3K, Akt and GLUT4 mRNA in skeletal muscles and adipose tissues. The results suggest that PPSC possess significant anti-diabetic activity, as evaluated using alloxanised diabetic mice model. Consequently, PPSC might be a promising candidate for the development of a new anti-diabetic agent.

Effects of Maydis stigma polysaccharide on the intestinal microflora in type-2 diabetes

Context: Diabetes is a serious endocrine and metabolic disorder. Food supplements attract people’s attention in mitigating health problems from the aspect of gastrointestinal microflora. Maydis stigma (Zea mays subsp. mays L. [Poaceae]), has been used as water decoction for treating diabetes in folk medicine. It has great potential, and feasibly a stable form of Maydis stigma commercial products could be developed to fulfil the health food market.Objective: To study the effects of Maydis stigma polysaccharide (MSP) on the intestinal microflora in type-2 diabetes (T2D).Materials and methods: MSP was fractioned from Maydis stigma by distilled water, purified by DEAE-52 Cellulose chromatography and Sephadex G-200 gel column. Streptozotocin (160 mg/kg) was intraperitoneal injected for 3 days to build model. The diabetic mice were randomly divided into five groups together with control group (10 mice in each group). The doses of MSP were 400, 600 and 800 mg/kg, respectively. After 5 weeks of administration, antidiabetic effects and intestinal microflora balance restoring activities were evaluated by denaturing gradient gel electrophoresis.Results: Blood glucose levels of MSP-treated groups showed extremely significant hypoglycemic effects (p < 0.01), body weight increased showed extremely significant (p < 0.01) differences. Bacteroides, Lactobacillus and Prevotella were dominant organisms in the intestinal tract. The quality and quantity of Lactobacillus and Bacteroides genus increased remarkably with increasing concentration of MSP.Discussion and conclusion: Experimental results of this study suggest that MSP has the significant potential to be used as a natural agent for treating T2D and restoring the intestinal microflora balance.

Isolation, Purification and Characterization of Lipase from Microbacterium sp. and its Application in Biodiesel Production

Presently the world is facing the problem of scarcity of fossil fuel as it is a non-renewable source of energy. Biodiesel can be an alternative energy source having the advantage of being renewable as well as environment friendly. Enzymes are better catalysts for the production biodiesel as enzymes are more stable and their production is more convenient and safer. Microalgae Scenedesmus sp. was used as a feedstock for biodiesel production. Lipase producing bacteria was isolated from the pulp and paper mill and identified as Microbacterium sp. using 16S rDNA sequencing method. Lipase enzyme was purified by sequential methods of ammonium sulphate precipitation and Sephadex G-100 gel column chromatography. The molecular weight of purified enzyme was 40 kDa on SDS-PAGE. This purification procedure resulted in 2.1fold purification of lipase with a 20.8% final yield. The purified lipase exhibited maximal hydrolytic activity at a temperature of 50 0C and a pH of 8.5. The Km of lipase was 3.2mM and the Vmax 50μmol/min/mg. Lipase activity was stimulated by Triton X-100 and SDS and inhibited by Tween 20 and Tween 80. Biodiesel was prepared through sodium hydroxide, potassium hydroxide and Lipase (Celite and charcoal bound as substrate) catalyzed transesterification process, which enabled a yield of 72.5%, 90% (95.1%, and 15.5%) respectively determined by gas chromatography/mass spectrometry (GC/MS) analysis.

Separation, purification and preliminary structure analysis of acidic polysaccharides from safflower

The crude water-soluble polysaccharide (SCP) was extracted from safflower by using hot water, and further fractioned by DEAE-52 anion exchange and Sephadex G-100 gel column chromatography to yield two acidic polysaccharides (SCP2 and SCP3). High-performance gel permeation chromatograph analysis showed that molecular weights (M W) of polysaccharides (SCP2 and SCP3) were 5, 860 and 9,330 Da, respectively. According to Fourier transform infrared spectroscopy, gas chromatography and chemical methods, SCP2 consisted of rhamnose, arabinose, glucose and galactose with a molar ratio of 2.93:11.19:33.68:3.48, while the primary sugar residues were (1 → 2)-linked-rhamnopyranose, (1 → 2)-linked and (1 → 3)-linked-arabinofuranose, (1 → 3)-linked-glucopyranose and (1 → 2)-linked or (1 → 6)-linked-galactopyranose. Likewise, SCP3 was composed of rhamnose, arabinose, xylose, mannose, glucose and galactose in a molar ratio of 4.44:1.46:4.51:5.82:8.23:19.38, while the primary sugar residues were (1 → 2)-linked-rhamnopyranose, (1 → 2)-linked-xylofuranose, (1 → 3)-linked-arabinofuranose, (1 → 2)-linked or (1 → 6)-linked-mannopyranose and glucopyranose, (1 → 2)-linked or (1 → 6)-linked or (1 → 3)-linked-galactopyranose.