Purification and Structure Study on Exopolysaccharides Produced by Lactobacillus paracasei KL1-Liu from Tibetan Kefir

2013 ◽  
Vol 781-784 ◽  
pp. 1513-1518 ◽  
Author(s):  
Hui Liu ◽  
Yuan Hong Xie ◽  
Tao Han ◽  
Hong Xing Zhang
Keyword(s):  
Molecular Weight ◽  
High Performance ◽  
Monosaccharide Composition ◽  
Gradient Elution ◽  
Structure Study ◽  
Lactobacillus Paracasei ◽  
Molar Ratio ◽  
Test Results ◽  
Sample Concentration ◽  
Multi Level

To investigate the pure conditions of exopolysaccharides produced by Lactobacillus paracasei KL1-Liu from Tibetan Kefir, and to analyse the structure, we used Multi-level single-factor test to purify EPS by Sepharose CL-6B. And the purity of EPS was detected by UV scan and high performance liquid chromatogram (HPLC). EPS molecular weight and monosaccharide composition were determined by HPLC. Results: Adoption phosphate buffer gradient elution 0.02-0.10 mol/L, the velocity 0.25 mL/min, the sample concentration 1.0 mg/mL, the sample capacity 1.0 mL. Under this purification conditions, components EPSa and EPSb were obtained. The purities of EPSa and EPSb were 82.82% and 91.74% respectively, which were 1.4 and 1.5 times of the pre-purification. Purity Test results showed that EPSa and EPSb polysaccharide were single components, basically no nucleic acid and protein in them. Structural analysis revealed that the molecular weight of EPSa and EPSb were 4.60×104 Da and 2.12×104 Da detected by HPLC. EPSa monosaccharide components were glucose and rhamnose, and the molar ratio was 1:0.68. EPSb were composed of glucose, xylose and rhamnose, and the molar ratio was 1:0.77:0.69.

scholarly journals Homologous Over-Expression of Chain Length Determination Protein EpsC Increases the Molecular Weight of Exopolysaccharide in Streptococcus thermophilus 05-34

2021 ◽  
Vol 12 ◽  
Author(s):  
Zhengyuan Zhai ◽  
Shuxin Xie ◽  
Hongxing Zhang ◽  
Huaxi Yi ◽  
Yanling Hao
Keyword(s):  
Molecular Weight ◽  
Chain Length ◽  
High Performance ◽  
Gel Permeation Chromatography ◽  
Streptococcus Thermophilus ◽  
Monosaccharide Composition ◽  
Molar Ratio ◽  
Gas Chromatography Mass Spectrometry ◽  
Over Expression ◽  
Length Determination

In Streptococcus thermophilus, EpsC is a polysaccharide co-polymerase which is involved in determining the chain length of EPS synthesized by the Wzx/Wzy-dependent pathway. Our previous study found that there was a positive correlation between transcription level of epsC and molecular weight of EPS in S. thermophilus 05-34. To further investigate the effects of EpsC on EPS biosynthesis, this gene was over-expressed in S. thermophilus 05-34 in this study. Reverse transcription qPCR and Western blotting confirmed the successful transcription and translation of epsC in 05-34, respectively. The yield of EPS was not affected by the over-expression of EpsC. Gas chromatography-mass spectrometry (GC-MS) showed that the monosaccharide composition was still composed of galactose and glucose in a molar ratio of 1.0:0.8, whereas high performance gel permeation chromatography (HPGPC) indicated that the molecular weight of EPS was increased from 4.62 × 105 Da to 9.17 × 105 Da by the over-expression of EpsC. In addition, S. thermophilus 05epsC which could produce higher molecular weight EPS improved the viscoelasticity and water-holding capacity of yogurt, but significantly reduced the level of syneresis in yogurt. In summary, these results indicated that homologous over-expression of EpsC in S. thermophilus could increase the molecular weight of EPS and improve the microrheological or physical properties of yogurt.

scholarly journals Physicochemical Characterization and Immunomodulatory Activity of a Novel Acid Polysaccharide from Solanum muricatum

Polymers ◽  
2019 ◽  
Vol 11 (12) ◽  
pp. 1972 ◽  
Author(s):  
Heng Yue ◽  
Qianqian Xu ◽  
Xianheng Li ◽  
Jeevithan Elango ◽  
Wenhui Wu ◽  
...  
Keyword(s):  
High Performance ◽  
Average Molecular Weight ◽  
Thermal Studies ◽  
Physicochemical Characterization ◽  
Monosaccharide Composition ◽  
Molar Ratio ◽  
Immunomodulatory Activity ◽  
Composition Analysis ◽  
Solanum Muricatum ◽  
Acid Polysaccharide

To investigate the structure and immunomodulatory activity of polysaccharide from Solanum muricatum, a novel acid polysaccharide named SMP-3a was purified from Solanum muricatum pulp through DEAE-52 cellulose column and Sephadex G-200 chromatography. Monosaccharide composition analysis showed that SMP-3a was mainly composed of rhamnose, arabinose, galactose, and galacturonic acid with the molar ratio of 1.09:2.64:1.54:1. The average molecular weight was found to be 227 kDa by high performance gel permeation chromatography (HPGPC). Thermal studies revealed the SMP-3a was a thermally stable polymer. Based on the results of methylation and NMR analysis, the backbone chain of SMP-3a was composed of →2)-α-l-Rhap-(1→, →4)-α-d-GalpA-(1→ and →4)-α-d-Galp-(1→. The side chain was consisted of α-l-Araf-(1→ and →5)-α-l-Araf-(1→. Immunomodulatory assay indicated that SMP-3a could significantly promote the proliferation of macrophages and stimulate the secretion of cytokines, including TNF-α, IL-1β, and IL-6. Our results suggested that SMP-3a could be used as a novel potential immunomodulatory agent in functional food.

scholarly journals Purification and Characterization of a New Lectin from Loach Skin Mucus

2019 ◽  
Vol 2019 ◽  
pp. 1-11
Author(s):  
Peng-peng Sun ◽  
Yuan-yuan Ren ◽  
Jie Zheng ◽  
Ai-jun Hu
Keyword(s):  
Molecular Weight ◽  
Carbohydrate Chain ◽  
Acid Resistance ◽  
Monosaccharide Composition ◽  
Molar Ratio ◽  
Pathogen Defense ◽  
Hemagglutination Activity ◽  
Purification And Characterization ◽  
Skin Mucus

Lectin from loach skin mucus plays an important role in pathogen defense. However, hardly can any paper relevant to the character of lectin from loach skin mucus be found in recent years. In this study, a kind of new lectin (LML), with a high hemagglutination activity of 166.23 × 103 HU/mg, was successfully isolated and purified from loach skin mucus. LML was a kind of glycoprotein with a molecular weight of 245 kDa. Also, the monosaccharide composition suggested that its carbohydrate chain was composed of rhamnose, arabinose, xylose, mannose, glucose, and galactose with a molar ratio of 2.02 : 11.66 : 2.06 : 1.00 : 14.09 : 6.00. Besides, LML depended on Ca2+ to induce hemagglutination and was strongly inhibited by D-lactose. The lectin exhibited powerful resistance to alkali and kept about 30% hemagglutination activity at pH 14.0, whereas its capacity of acid resistance was weak. The maximum hemagglutination activity of LML maintained at a temperature range from 20°C to 50°C. Moreover, the structure of LML was preliminarily studied, indicating it contained abundant glutamic acid, histidine, and serine, and its secondary structure contained α-helix (4.97%), β-sheet (27.55%), turns structure (49.78%), and unordered structure (17.70%).

Production and Rheological Properties of Welan Gum Produced by Sphingomonas sp. ATCC 31555 with Different Nitrogen Sources

2017 ◽  
Vol 27 (1) ◽  
pp. 55-63 ◽  
Author(s):  
Xiaopeng Xu ◽  
Zuoming Nie ◽  
Zhiyong Zheng ◽  
Li Zhu ◽  
Xiaobei Zhan
Keyword(s):  
Molecular Weight ◽  
Rheological Properties ◽  
Biomass Concentration ◽  
Bacterial Biomass ◽  
Nitrogen Sources ◽  
Monosaccharide Composition ◽  
Solution Concentration ◽  
Molar Ratio ◽  
Welan Gum ◽  
Beef Extract

This study aimed to investigate the effect of nitrogen sources on the production and rheological properties of welan gum produced by <i>Sphingomonas</i> sp. ATCC 31555. Six different nitrogen sources were used for ATCC 31555 fermentation, and 2 of these were further analyzed due to their more positive influence on welan gum production and bacterial biomass. Bacterial biomass, welan gum yield, welan viscosity, molecular weight, monosaccharide composition, acyl content, and welan structure were analyzed. Welan gum production and the biomass concentration of ATCC 31555 were higher in media containing NaNO<sub>3</sub> and beef extract. Welan viscosity decreased at higher temperatures of 30-90°C, and it increased with a higher welan concentration. In the media containing NaNO<sub>3</sub> (3 g·L<sup>-1</sup>), welan viscosity was higher at 30-70°C and a welan solution concentration of 6-10 g·L<sup>-1</sup>. With a reduced NaNO<sub>3</sub> concentration, the molecular weight of welan gum and the molar ratio of mannose decreased, but the molar ratio of glucuronic acid increased. With different nitrogen sources, the acetyl content of welan gum differed but its structure was similar. NaNO<sub>3</sub> and beef extract facilitated welan production. A reduced NaNO<sub>3</sub> concentration promoted welan viscosity.

scholarly journals A Sensitive and Rapid UPLC-MS/MS Method for Determination of Monosaccharides and Anti-Allergic Effect of the Polysaccharides Extracted from Saposhnikoviae Radix

Molecules ◽  
2018 ◽  
Vol 23 (8) ◽  
pp. 1924 ◽  
Author(s):  
Yan-Yan Gao ◽  
Yue Jiang ◽  
Guo-Chao Chen ◽  
Shuang-Shuang Li ◽  
Fei Yang ◽  
...  
Keyword(s):  
High Performance ◽  
Monosaccharide Composition ◽  
Water Soluble ◽  
Delayed Type Hypersensitivity ◽  
Molar Ratio ◽  
Serum Ige ◽  
Organ Index ◽  
Wide Range ◽  
Liquid Chromatography Tandem Mass ◽  
Saposhnikoviae Radix

Background: Allergic disease is a common clinical disease. Natural products provide an important source for a wide range of potential anti-allergic agents. This study was designed to evaluate the anti-allergic activities of the water-soluble polysaccharides extracted and purified from Saposhnikoviae Radix (SRPS). The composition and content of monosaccharides were determined to provide a material basis. Methods: An ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established to determine the composition and content of SRPS. 2,4-dinitrofluorobenzene (DNFB) induced a delayed-type hypersensitivity (DTH) mouse model orally administrated SRPS for seven consecutive days. Ear swelling, organ index, and serum IgE levels were observed to evaluate the anti-allergic activities. Results: The UPLC-MS/MS analysis showed that SRPS was consisted of eight monosaccharides including galacturonic acid, mannose, glucose, galactose, rhamnose, fucose, ribose, and arabinose with a relative molar ratio of 4.42%, 7.86%, 23.69%, 12.06%, 3.10%, 0.45%, 0.71%, and 47.70%, respectively. SRPS could effectively reduce ear swelling, a thymus index, and a serum IgE levels. Conclusions: The method was simple, rapid, sensitive, and reproducible, which could be used to analyze and determine the monosaccharide composition of SRPS. The vivo experiments demonstrated that SRPS may effectively inhibit development of DNFB-induced DTH. SRPS is a novel potential resource for natural anti-allergic drugs.

scholarly journals Determination of 25 Low Molecular Weight Carbohydrates in Tobacco by High Performance Ion Chromatography

1994 ◽  
Vol 16 (2) ◽  
pp. 77-84 ◽  
Author(s):  
MC Ramusino ◽  
BS Dattilo ◽  
A Lucibello ◽  
SG Rossi
Keyword(s):  
Molecular Weight ◽  
High Performance ◽  
Chromatographic Method ◽  
Gradient Elution ◽  
Low Molecular Weight ◽  
Qualitative And Quantitative ◽  
Naoh Solution ◽  
Modified Polystyrene ◽  
Isocratic And Gradient Elution

AbstractA High Performance Ion Chromatographic method for the qualitative and quantitative determination of low molecular weight carbohydrates in tobacco leaf is described. The carbohydrates were separated by running isocratic or gradient elutions on columns packed with amino-modified polystyrene divinylbenzene phases, using 0.1 N NaOH as eluent. The compounds were detected by a Pulsed Amperometric Detector, after post-column addition of 1 N NaOH solution. Twenty-two and twenty-four carbohydrates out of twenty-five were separated by isocratic and gradient elution, respectively. Some mono and disaccharides were identified in tobacco samples by their retention times and some quantitative determinations of glucose, fructose, inositol and saccharose were carried out using different varieties of tobacco.

scholarly journals A Polysaccharide Produced by Lactococcus lactis subsp. lactis YZ1 Isolated from Traditional Indonesian Fermented Milk, "Dadih"

2000 ◽  
Vol 1 (1) ◽  
pp. 1-5
Author(s):  
Yusdar Zakaria
Keyword(s):  
Molecular Weight ◽  
Ion Exchange ◽  
Lactococcus Lactis ◽  
Monosaccharide Composition ◽  
Fermented Milk ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Molar Ratio ◽  
Neutral Sugar ◽  
Glycoside Linkage

ABSTRACT.Lactococcus lactis subsp. Lactis YZI was isolated from M17 agar in which diluted Dadih was poured and incubated at 30 0C for 48 h. Taxonomix properties of the isolate were examined according to Bergey’s Manual of Systematic Bacteriologi and Manual for  Identification of Medical Bacteria. The isolation of polysaccharide from the precipitant was performed on an ion-exchange chromatography. The result showed that the polysaccharides produced by Lactococus lactis subsp. lactis YZI were neutral sugar (unadsorbrd fraction) and glycoconjugated (absorbed fraction). The neutral sugar had molecular weight of 10,000 and 20,000 with and α-glycoside linkage. The monosaccharide composition was mannose, glucose and galactose with a molar ratio of 1 :1,5 : 4,9.

scholarly journals Characterization and Antioxidant Activity of a Low-Molecular-Weight Xanthan Gum

Biomolecules ◽  
2019 ◽  
Vol 9 (11) ◽  
pp. 730 ◽  
Author(s):  
Hu ◽  
Wang ◽  
Yu ◽  
He ◽  
Qiao ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Hydroxyl Radicals ◽  
Xanthan Gum ◽  
Glucuronic Acid ◽  
Reducing Power ◽  
Monosaccharide Composition ◽  
Molar Ratio ◽  
Low Molecular Weight ◽  
Oxygen Species ◽  
Superoxide Anions

In the present work, a low-molecular-weight xanthan gum (LW-XG) was successfully obtained via biodegradation of commercial xanthan by the endophytic fungus Chaetomium globosum CGMCC 6882. The monosaccharide composition of LW-XG was glucose, mannose, and glucuronic acid in a molar ratio of 1.63:1.5:1.0. The molecular weight of LW-XG was 4.07 × 104 Da and much smaller than that of commercial xanthan (2.95 × 106 Da). Antioxidant assays showed that LW-XG had a good scavenging ability on DPPH radicals, superoxide anions, and hydroxyl radicals and good ferric reducing power. Moreover, LW-XG exhibited excellent protective effect on H2O2-injured Caco-2 cells. Results of this work suggested that LW-XG could be used in foods or pharmaceuticals to alleviate and resist the oxidative damage induced by the overproduction of reactive oxygen species.

scholarly journals Research of Inonotus obliquus Oligosaccharide in Prevention of Hyperlipidemia

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Dawei Wu ◽  
Yanrong Zhang ◽  
Dawei Wang ◽  
Tingting Liu ◽  
Shanshan Zhang ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Protective Effect ◽  
Intestinal Flora ◽  
Deae Cellulose ◽  
Monosaccharide Composition ◽  
Hot Water ◽  
Molar Ratio ◽  
Inonotus Obliquus ◽  
Glycosidic Bonds ◽  
Liver And Kidney

In this study, hot water was used to extract Inonotus obliquus oligosaccharide. DEAE-cellulose and Sepharose G-200 were used to purify Inonotus obliquus oligosaccharide. Inonotus obliquus oligosaccharide IOP-2A was obtained. Its molecular weight Mw is about 1000 Da. The monosaccharide composition and molar ratio were glucose : xylose : galactose : mannose = 54.1 : 13.6 : 13.2 : 6.7. In addition, it also contains a small amount of galactose, gluconic acid, rhamnose, and fucose. IOP-2A contained mainly β-glycosidic bonds. Among them, 1,4-glycosidic bonds accounted for 9.2%, and 1,6-glycosidic bonds accounted for 85.1%. Oligosaccharide macromolecules formed a layered structure. Mouse experiments showed that IOP-2A had the function of preventing hyperlipidemia. At the same time, IOP-2A had a certain protective effect on the liver and kidney. The mechanism of IOP-2A in preventing hyperlipidemia was obtained from the perspective of mouse intestinal flora.

Composition Distribution of Acrylonitrile-Butadiene Copolymers as Analyzed by High-Performance Liquid Chromatography

1990 ◽  
Vol 63 (2) ◽  
pp. 181-190 ◽  
Author(s):  
N. Asada ◽  
H. Hosozawa ◽  
A. Toyoda ◽  
H. Sato
Keyword(s):  
Molecular Weight ◽  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Chemical Composition ◽  
Stationary Phase ◽  
High Performance ◽  
Adsorption Mechanism ◽  
Gradient Elution ◽  
Composition Distribution

Abstract The separation of acrylonitrile-butadiene copolymer (NBR) was investigated by high-performance liquid chromatography (HPLC). The copolymer was most successfully separated on the basis of chemical composition by using crosslinked 2-chloroacrylonitrile gel as the stationary phase and with a gradient elution of chloroform/n-hexane. The separation was found to be carried out by an adsorption mechanism and essentially independent of the molecular weight of the sample. The observed chemical composition distributions (CCD) agreed with those calculated from the copolymerization theory.

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