Cellulose Column Research Articles

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Strain Enterococcus sp. GHB26, isolated from the Algerian paste of dates Ghars, produced a bacteriocin. This bacteriocin was inactivated by proteolytic enzymes. Antibacterial activity of the bacteriocin was heat stable at 120°C for 20 min (533 AU/ml), stable at pH range of 2 to 12 and resistant to chemicals (SDS, EDTA, NaCl, Tween 80, Urea). The active bacteriocin from the cell-free supernatant of Enterococcus sp. GHB26 was purified by precipitation with ammonium sulfate followed by various combinations of gel filtration on a Sephadex G-25 column, cation exchange chromatography on a CM- Sephadex Cellulose column and reverse phase-high performance liquid chromatography on a C18 column. The bacteriocin was eluted as a single peak on the chromatogram from reverse phase-high performance liquid chromatography attesting the purity of this bacteriocin. The bacteriocin exhibited a bactericidal mode of action. Sodium dodecyl sulphate - polyacrylamide gel electrophoresis indicated that molecular weight of this bacteriocin is close to 3.5 kDa. Key words: Paste of dates, bacteriocin, antibacterial activity, chromatography, mode of action.

Extraction optimization, preliminary characterization and immunological activities in vitro of polysaccharides from Elaeagnus angustifolia L. pulp

In this research, extraction optimization, preliminary characterization and immunological activities in vitro of polysaccharides from Elaeagnus angustifolia L. pulp were investigated. A response surface methodology (RSM) with a Box-Behnken design (BBD) was used to optimize the extraction process. The maximum EAP yield was 9.82±0.38%, which is in good agreement with the predicted value (9.93±0.24%). Two homogeneous polysaccharides, EAP-1a and EAP-1b with molecular weights of 8.70kDa and 4.39kDa respectively, were prepared by DEAE-52 cellulose and Sephadex G-100 columns and characterized by HPLC, HPGPC, and FT-IR. Three polysaccharides (EAP, EAP-1a and EAP-1b) could stimulate macrophages to release NO and enhance phagocytic activities of RAW 264.7 cells in dose-dependent manner. Moreover, there was no significant difference between crude EAP group (400μg/mL) and positive control group (LPS) in effects on macrophages. The results implied that EAP had the potential to be developed as natural medicines or health foods.

Optimization of infrared-assisted extraction of Bletilla striata polysaccharides based on response surface methodology and their antioxidant activities

Bletilla striata polysaccharides (BSP) have attracted extensive research interest due to their potential medical application. Herein, infrared-assisted technique is employed for the first time to extract BSP from B. striata (Thunb.) Reichb.f. based on a Box-Behnken design (BBD) and response surface methodology, with the optimum extraction parameters as follows: 75°C extraction temperature, 2.5h extraction time; and water to solid ratio (53ml/g). Based on it, 43.95±0.26% yield of crude BSP was obtained. Subsequently, crude BSP was further decolorized, deproteinized, freeze-dried, and purified by a DEAE-52 cellulose column. Furthermore, the micro-structure and a triple-helical structure of BSP were characterized. Fourier transform infrared spectra confirmed its polysaccharide characterization via typical peaks. In addition, the significant in vitro antioxidant profiles of BSP were demonstrated by superoxide anion radical-scavenging assay, hydroxyl radical scavenging assay, DPPH free radical scavenging activity and chelation of ferrous ions. Taken together, this study provide an efficient extraction technique for BSP as a promising natural antioxidant.

Antioxidant activity of Inonotus obliquus polysaccharide and its amelioration for chronic pancreatitis in mice

Inonotus obliquus polysaccharide (IOP) was extracted by water with a yield of 9.83% and purified by an anion-exchange DEAE cellulose column and Sephadex G-200 gel with a polysaccharide content of 98.6%. The scavenging activities for 2,2-diphenyl-1-picryl-hydrazyl (DPPH) and hydroxyl radicals of IOP were 82.3% and 81.3% respectively at a concentration of 5 mg/mL. IOP was composed of Man, Rha, Glu, Gal, Xyl and Ara in a molar ratio of 9.81:3.6：29.1：20.5：21.6：5.4 respectively. The gel permeation chromatography indicated that IOP was a homogeneous polysaccharide with molecular weight of 32.5 kDa. IOP helped to alleviate pancreatic acinar atrophy and weight loss for chronic pancreatitis (CP) mice induced by Diethyldithiocarbamate (DDC). The SOD level was increased most by IOP-H treatment (400 mg/kg body weight). MDA, IL-1β and LDH were significantly decreased by IOP treatment, especially hydroxyproline, IFN-γ and AMS levels were decreased 39.18%, 37.82% and 41.57% by IOP-H treatment respectively compared to MC group. In conclusion, IOP possessed strong antioxidant activity for scavenging free radicals in vitro and vivo which could be propitious to CP therapy in mice.

The role of lipoprotein(a) in clotting reactions during lipoprotein apheresis—A case report

In individuals with familial hypercholesterolemia (FH) who are unable to reach a target low-density lipoprotein level on a drug regimen, lipoprotein apheresis (LA) may be the treatment of choice. Severe reactions involving clotting during LA are not well described in the literature. We report a case of a 63-year-old woman with FH and markedly elevated lipoprotein(a) (Lp[a]) levels who experienced such a reaction while undergoing LA with a dextran-sulfate cellulose column on the Kaneka MA-01 Liposorber system. Owing to the clotting as well as a blood pressure drop to <100 mm Hg systolic, the procedure was stopped early. Before her second procedure, she was given an increased loading dose of unfractionated heparin. She did not develop clotting during this second procedure. A growing body of literature on the role of Lp(a) in atherothrombotic complications and hemostasis supports a possible mechanism by which clotting in the instrument could occur during apheresis. Our patient's initial pretreatment Lp(a) was 3.5 times greater than the mean Lp(a) levels in patients with FH. This theory is consistent with our case in that the patient's Lp(a) levels progressively declined with each procedure, and she had no subsequent clotting.

Partial Purification and Thermal Stability of Two Peroxidases from Pithecellobium dulce (Roxb.) Benth. Aril

Two peroxidase (POX) forms, named PdPI and PdPII, were partially purified from the Pithecellobium dulce aril, using ammonium sulfate precipitation and anion exchange chromatography. Zymography of the proteins obtained after precipitation in the range of 60-90% ammonium sulfate (F6090 fraction) showed two bands with peroxidase activity. In DE-52 cellulose column, the peroxidase activity was detected in unadsorbed peak (PdPI) and adsorbed peak eluted with 0.2 mol L -1 NaCl (PdPII) suggesting that PdPI (~114 kDa) and PdPII (~77 kDa) are basic and acidic proteins, respectively, being PdPI stable at 80oC. This is the first report of partial purification of POX from P. dulce aril, indicating that this species may be a new source of enzymes for use in biosensors and other biotechnological applications.

Ultrasonic extraction, antioxidant and anticancer activities of novel polysaccharides from Chuanxiong rhizome

Ultrasonic-assisted extraction technology was employed to prepare Ligusticum chuanxiong Hort polysaccharide. Single factor test and orthogonal experimental design were used to optimize the extraction conditions. The results showed that the optimal extraction conditions consisted of ultrasonic temperature of 80°C, ultrasonic time of 40min and water to raw material ratio of 30mL/g. Three novel polysaccharides fractions, LCX0, LCX1 and LCX2, were isolated and purified from the crude polysaccharides using DEAE-52 cellulose and Sephadex G-100 column chromatography. The molecular weight and monosaccharide composition of three LCX polysaccharides fractions were analyzed with gel permeation chromatography (GPC) and HPLC analysis, respectively. Furthermore, the antioxidant and in vitro anticancer activities of the polysaccharides were investigated. Compared with LCX0, LCX2 and LCX1 showed relative higher antioxidant activity and inhibitory activity to the growth of HepG2, SMMC7721, A549 and HCT-116 cells. It is suggested that the novel polysaccharides from rhizome of L. chuanxiong could be promising bioactive macromolecules for biomedical use.

Evaluation of bioremediation potentiality of ligninolytic Serratia liquefaciens for detoxification of pulp and paper mill effluent

Due to high pollution load and colour contributing substances, pulp and paper mill effluents cause serious aquatic and soil pollution. A lignin-degrading bacterial strain capable of decolourising Azure-B dye was identified as lignin peroxidase (LiP) producing strain LD-5. The strain was isolated from pulp and paper mill effluent contaminated site. Biochemical and 16S rDNA gene sequence analysis suggested that strain LD-5 belonged to the Serratia liquefaciens. The strain LD-5 effectively reduced pollution parameters (colour 72%, lignin 58%, COD 85% and phenol 95%) of real effluent after 144h of treatment at 30°C, pH 7.6 and 120rpm. Extracellular LiP produced by S. liquefaciens during effluent decolourisation was purified to homogeneity using ammonium sulfate (AMS) precipitation and DEAE cellulose column chromatography. The molecular weight of the purified lignin peroxidase was estimated to be ∼28kDa. Optimum pH and temperature for purified lignin peroxidase activity were determined as pH 6.0 and 40°C, respectively. Detoxified effluent was evaluated for residual toxicity by alkaline single cell (comet) gel electrophoresis (SCGE) assay using Saccharomyces cerevisiae MTCC 36 as model organism. The toxicity reduction to treated effluent was 49.4%. These findings suggest significant potential of S. liquefaciens for bioremediation of pulp and paper mill effluent.

Separation, characterization and anticancer activities of a sulfated polysaccharide from Undaria pinnatifida

The purpose of this paper was to investigate separation, characterization and anticancer activities of a sulfated polysaccharide (SPUP) from Undaria pinnatifida. Firstly, polysaccharide from U. pinnatifida was separated by DEAE-52 cellulose and Sephacryl S-400 column chromatography. As results, SPUP was obtained with the yield of 19.42%. Then, SPUP was characterized using chemical analysis, gas chromatography, size-exclusion HPLC chromatography, UV–vis spectra and FT-IR spectrum. The content of total sugar, uronic acid, protein and sulfate radical were 80.48%, 3.21%, 7.12% and 29.14%, respectively. SPUP was a heteropolysaccharide composed of fucose, glucose and galactose in a molar percentage of 27.15:19.34:53.51 with molecular weight of 97.9kDa. Finally, the strongly against breast cancer activity of SPUP was confirmed by DMBA-induced breast cancer rats model. AS results, SPUP can significantly restrain breast abnormal enlargement, prolong tumor latency and reduced tumor incidence. Immunomodulatory activity and regulating abnormal sex hormones level might contribute to its anticancer activities.

Dimeric Octaketide Spiroketals from the Jellyfish-Derived Fungus Paecilomyces variotii J08NF-1.

Paeciloketals (1-3), new benzannulated spiroketal derivatives, were isolated from the marine fungus Paecilomyces variotii derived from the giant jellyfish Nemopilema nomurai. Compound 1 was present as a racemate and was resolved into enantiopure 1a and 1b by chiral-phase separation on a cellulose column. Compounds 2 and 3, possessing a novel benzannulated spiroketal skeleton, were rapidly interconvertible and yielded an equilibrium mixture on standing at room temperature. The relative and absolute configurations of compounds 2 and 3 were determined by NOESY analysis and ECD calculations. Compound 1 showed modest antibacterial activity against the marine pathogen Vibrio ichthyoenteri.

Novel functional polysaccharides from Radix Polygoni Multiflori water extracted residue: Preliminary characterization and immunomodulatory activity

The alkali-extractable polysaccharides (APMPs) were isolated from the water extracted residues of Radix Polygoni Multiflori, and further purified by DEAE-52 cellulose and Sephadex G-100 column chromatography to obtain a homogeneous polysaccharide (APMP-2) with molecular weights of 7724.8Da. HPLC chromatography analysis identified that APMP-2 was a heteropolysaccharides and mainly composed of Galactose and Xylose with a molar ratio of 4.31: 1.06. It was shown that both APMP and APMP-2 were of activation effects on splenocytes and peritoneal macrophages, and also significantly restore the proliferation rate, phagocytic index and cytokine (IL-2 and TNF-α) production level of 5-FU-treated splenocytes/peritoneal macrophages in a dosage-dependent manner. The results suggested that polysaccharides presented in Radix Polygoni Multiflori water-extracted residues possessed immunomodulatory activity and could be used as potential immunomodulators, and this finding could be a reference for the utilization of Radix Polygoni Multiflori water extracted residues.

Regenerated bacterial cellulose microfluidic column for glycoproteins separation

To analysis and separate glycoproteins, a simple strategy to prepare regenerated bacterial cellulose (RBC) column with concanavalin A (Con A) lectin immobilized in microfluidic system was applied. RBC was filled into microchannel to fabricate RBC microcolumn after bacterial cellulose dissolved in NaOH–sulfourea water solution. Lectin Con A was covalently connected onto RBC matrix surface via Schiff-base formation. Lysozyme (non-glycoprotein) and transferrin (glycoprotein) were successfully separated based on their different affinities toward the immobilized Con A. Overall, the RBC microfluidic system presents great potential application in affinity chromatography of glycoproteins analysis, and this research represents a significant step to prepare bacterial cellulose (BC) as column packing material in microfluidic system. What is more, troublesome operations for lectin affinity chromatography were simplified by integrating the microfluidic chip onto a HPLC (High Performance Liquid Chromatography) system.

Optimization of extraction, characterization and antioxidant activity of polysaccharides from Brassica rapa L.

The root of Brassica rapa L. has been traditionally used as a Uyghur folk medicine to cure cough and asthma by Uyghur nationality in Xinjiang Uygur Autonomous Region of China. In the present study, therefore, extraction optimization, characterization and antioxidant activity in vitro of polysaccharides from the root of B. rapa L. (BRP) were investigated. The optimal extraction conditions with an extraction yield of 21.48±0.41% for crude BRP were obtained as follows: extraction temperature 93°C, extraction time 4.3h and ratio of extraction solvent (water) to raw material 75mL/g. The crude BRP was purified by chromatographic columns of DEAE-52 cellulose and Sephadex G-100, affording three purified fractions of BRP-1-1, BRP-2-1 and BRP-2-2 with average molecular weight of 1510, 1110 and 838kDa, respectively. Monosaccharide composition analysis indicated that BRP-1-1 was composed of mannose, rhamnose, glucose, galactose and arabinose, BRP-2-1 was composed of rhamnose, galacturonic acid, galactose and arabinose, and BRP-2-2 was composed of rhamnose and galacturonic acid in a molar ratio of 1.27: 54.92. Furthermore, the crude BRP exhibited relatively higher antioxidant activity in vitro than purified fractions; hence, it could be used as a natural antioxidant in functional foods or medicines.

Plasmodium falciparum merozoite surface protein 1 block 2 gene polymorphism in field isolates along the slope of mount Cameroon: a cross - sectional study.

BackgroundMalaria remains a major global health burden despite the intensification of control efforts, due partly to the lack of an effective vaccine. Information on genetic diversity in natural parasite populations constitutes a major impediment to vaccine development efforts and is limited in some endemic settings. The present study characterized diversity by investigating msp1 block 2 polymorphisms and the relationship between the allele families with ethnodemographic indices and clinical phenotype.MethodIndividuals with asymptomatic parasitaemia (AP) or uncomplicated malaria (UM) were enrolled from rural, semi-rural and semi-urban localities at varying altitudes along the slope of mount Cameroon. P. falciparum malaria parasitaemic blood screened by light microscopy was depleted of leucocytes using CF11 cellulose columns and the parasite DNA genotyped by nested PCR.ResultsLength polymorphism was assessed in 151 field isolates revealing 64 (5) and 274 (22) distinct recombinant and major msp1 allelic fragments (genotypes) respectively. All family specific allelic types (K1, MAD20 and RO33) as well as MR were observed in the different locations, with K1 being most abundant. Eighty seven (60 %) of individuals harbored more than one parasite clone, with a significant proportion (p = 0.009) in rural compared to other settings. AP individuals had higher (p = 0.007) K1 allele frequencies but lower (p = 0.003) mean multiplicity of genotypes per infection (2.00 ± 0.98 vs. 2.56 ± 1.17) compared to UM patients.ConclusionsThese results indicate enormous diversity of P. falciparum in the area and suggests that allele specificity and complexity may be relevant for the progression to symptomatic disease.

Purification, structural characterization and anticancer activity of the novel polysaccharides from Rhynchosia minima root

Three novel acidic polysaccharides termed PRM1, PRM3 and PRM5 were purified from Rhynchosia minima root using DEAE-52 cellulose and sephadex G-150 column chromatography. Their structures were characterized by ultraviolet (UV) and Fourier transform infrared (FTIR) spectrometry, gel permeation chromatography (GPC), gas chromatography–mass spectrometry (GC–MS), and differential scanning colorimeter (DSC) analysis. The uronic acid contents of PRM1, PRM3 and PRM5 were 30.7%, 12.7% and 47.7%, respectively. PRM1 (143.2kDa), PRM3 (105.3kDa) and PRM5 (162.1kDa) were heteropolysaccharides because they were composed of arabinose, mannose, glucose and galactose. Their enthalpy values were 201.0, 111.0 and 206.8J/g, respectively. PRM3 and PRM1 exhibited strong in vitro anticancer activity against lung cancer A549 and liver cancer HepG2 cells in a dose-dependent manner. These findings suggested that PRM1 and PRM3 could be potentially developed as natural anticancer agents.

Characterization and antitumor activities of a water-soluble polysaccharide from Ampelopsis megalophylla

A water-soluble polysaccharide named as AMP was isolated and purified from the leaves of Ampelopsis megalophylla by DEAE-52 Cellulose and Sephadex G-100 column chromatography. AMP had an average molecular weight of about 8.4×104Da and was composed of galactose (Gal), mannose (Man), glucose (Glc), arabinose (Ara), and rhamnose (Rha) in a molar ratio of 2.7:1.6:1.1:0.6:0.3. After 10 days of AMP (50, 100, and 200mg/kg) treatment once daily in tumor-bearing mice, AMP oral administration could inhibit the growth of transplantable Sarcoma 180 (S180) tumor in mice and increase the spleen index and body weight. Furthermore, AMP also promote splenocytes’ proliferation induced by concanavalin A (ConA) and lipopolysaccharide (LPS), strengthen peritoneal macrophages to devour neutral red and increase the production of interleukin-2 (IL-2), tumor necrosis factor-alpha (TNF-α), and interferon-gamma (IFN-γ) in serum. These results suggest that AMP had clear antitumor activity, which might be related to its regulation of immune function in mice.

Biochemical analysis and hypoglycemic activity of a polysaccharide isolated from the fruit of Lycium barbarum L

Purification, characterization and hypoglycemic activities of polysaccharide from Lycium barbarum L. were investigated in this study. A water soluble polysaccharide (LBP) was obtained with ultrafiltration membranes separation, which was further purified by chromatography of DEAE cellulose column and Sephadex G-150 to get LBP3a and LBP3b. The high performance permeation chromatography (HPGPC) analysis showed that the average molecular weight (Mw) of LBP3b was 4.92kDa. Monosaccharide composition analysis revealed that the LBP3b was comprised of mannose, rhamnose, glucose, galactose and xylose with a molar ratio of 5.52:5.11:28.06:1.00:1.70. The preliminary structure features of LBP3b were investigated by UV, FT-IR, NMR and SEM. In vitro cell experiments showed that LBP3b had significantly inhibited the absorption of glucose in a dose-dependent manner. The study showed that LBP3b had potential use as an anti-diabetic agent.

High-performance liquid chromatographic enantioseparation of unusual amino acid derivatives with axial chirality on polysaccharide-based chiral stationary phases

The successful enantioseparation of axially chiral amino acid derivatives containing a cyclohexylidene moiety on an analytical and semipreparative scale was achieved for the first time by HPLC using polysaccharide-based chiral stationary phases. Racemic methyl N-benzoylamino esters, easily obtained by methanolysis of the corresponding 5(4H)-oxazolones, were subjected to chiral HPLC resolution using chiral stationary phases based on immobilized 3,5-dimethylphenylcarbamate derivatives of amylose (Chiralpak® IA column) or cellulose (Chiralpak® IB column). The behaviour of both selectors under different elution conditions was evaluated and compared. The amylose column showed better performance than the cellulose column for all enantiomers tested. The semipreparative resolution of axially chiral amino acid derivatives with different side chains has been achieved on a 250mm×20mm ID Chiralpak® IA column using the appropriate mixture of n-hexane/chlorofom/ethanol as eluent by successive injections of a solution of the sample in chloroform. Using this protocol up to 120mg of each enantiomer of the corresponding axially chiral amino acid derivative were obtained from 300mg of racemate. [(Sa)-2a, 105mg; (Ra)-2a, 60mg, [(Sa)-2b, 105mg; (Ra)-2b, 90mg, [(Sa)-2c, 120mg; (Ra)-2c, 100mg].

A novel polysaccharide from mycelia of cultured Phellinus linteus displays antitumor activity through apoptosis

Two novel polysaccharides termed PLPS-1 and PLPS-2 were isolated from mycelia of cultured Phellinus linteus by hot water extraction, purified by DEAE-52 cellulose and Sephadex G-100 column chromatography, and structurally characterized by FTIR and NMR spectroscopy, GC–MS, periodate oxidation/Smith degradation, and methylation analysis. The monosaccharide compositions of PLPS-1 (MW 2.5×105Da) and PLPS-2 (MW 2.8×104Da) were respectively Glc, Ara, Fuc, Gal, and Xyl in molar ratio 21.964:1.336:1.182:1:1, and Glc, Gal, Man, Ara, Fuc, Xyl in molar ratio 14.368:2.594:1.956:1.552:1.466:1; i.e., both were heteropolysaccharides. The backbone of PLPS-1 consisted primarily of repeating α-d-Glc(1→4)-α-d-Glc(1→6) units, while that of PLPS-2 consisted of α-(1→3)-d-Glc and α-(1→6)-d-Glc. The side branches were also different in their carbohydrate components. In in vitro antitumor assays, PLPS-1 displayed strong anti-proliferative effect against S-180 sarcoma cells through apoptosis, whereas PLPS-2 had no such effect. The difference in antitumor activity between the two PLPS evidently results from their structural differences. PLPS-1 has potential as a novel anticancer agent.

A novel alkali extractable polysaccharide from Plantago asiatic L. Seeds and its radical-scavenging and bile acid-binding activities.

A new acidic polysaccharide (PLP) was isolated and characterized from Plantago asiatic L. seeds by hot alkali extraction and chromatographic purification using DEAE cellulose and Sephacryl S-400 columns. PLP has a molecular weight of 1.15 × 10(6) Da, and a monosaccharide composition of xylose (Xyl), arabinose (Ara), glucuronic acid (GlcA), and galactose (Gal) in a molar ratio of 18.8:7.2:6.1:1. The results of methylation analysis, FT-IR, and 1D and 2D NMR indicated that PLP was a highly branched heteroxylan of β-1,4-linked Xylp backbone with three α-GlcAp-(1→3)-Araf attached to the O-3 position and one α-T-linked-GlcAp and one α-Araf-(1→5)-Araf attached to the O-2 position every eight monosaccharide residues. PLP exhibited scavenging abilities against hydroxyl, peroxyl anion, and DPPH radicals in vitro and showed significant binding capacities against cholic and chenodeoxycholic acids, suggesting its possible cholesterol-lowering activity. The results demonstrated the potential use of PLP in functional foods and nutraceuticals.