1.Studies on separation, purification and structure characteristics of a polysaccharide LTC-II from Pyrola corbieri.
Zhengchang MO ; Lanfang WU ; Juan YANG ; Daoping WANG
China Journal of Chinese Materia Medica 2011;36(12):1633-1636
OBJECTIVETo characterize the structure of polysaccharide LTC-II obtained from Pyrola corbieri.
METHODThe polysaccharide was extracted from P. corbieri by hot water and ethanol precipitation. Crude polysaccharide was purified by DEAE-Cellulose chromatography and Sephacryl S-300 HR column chromatography. The purity and molecular weight of polysaccharide was determined by gel permeation chromatography. UV, IR, optical rotation, complete acid hydrolysis, periodate oxydation, Smith degradation, partial acid hydrolysis and methylation analysis were applied to determine the structural features.
RESULTA homogeneous fraction LTC-II was obtained and its relative molecular mass was 22 000 Da. It consisted of arabinose, mannose, glucose, galactose in the molar ratio of 35. 2: 1.0: 13. 4: 4. 2. LTC-II had a backbone consisting glucose, mannose, galactose and mainly contained (1 --> 6)-linkaged glucose. The side chain possessed arabinose, glucose, galactose and mainly contained (1 --> 5)-linkaged arabinose. The terminal sugar were mainly glucose and galactose.
CONCLUSIONStudies on the preliminary characterization of polysaccharide LTC-II from P. corbieri for the first time.
Chromatography ; Polysaccharides ; chemistry ; Pyrola ; chemistry
2.Rapid N-glycosylation profiling of glycoprotein based on ultrafiltration-devices.
Cai TIE ; Ting HU ; Jin-lan ZHANG
Acta Pharmaceutica Sinica 2015;50(6):755-759
With development of bio-technique, more and more proteins were applied as clinical approaches. However, the protein homogeneity, especially the N-glycosylation limited the further research and application of these protein drugs. The analysis method for N-glycans is believed to be critical in protein drugs development. To enhance the N-glycans isolation efficiency and accelerate the pretreatment, a new strategy was built on ultrafiltration-devices. New methods increased the isolation efficiency of N-glycans containing N-acetylglucosa mine with 10%-20%. The degrading of N-glycans containing sialic acids was also minimized with this method. 20%-100% more N-glycans with sialic acids were isolated. The pretreatment was finished within 30 min. Coupled with HPLC-HRMS, an effective and reliable strategy designed for protein drugs N-glycans analysis were developed.
Glycoproteins
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chemistry
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Glycosylation
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Polysaccharides
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chemistry
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Ultrafiltration
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instrumentation
3.Purification and characterization of extracellular polysaccharides from Nostoc flagelliforme.
Chinese Journal of Biotechnology 2008;24(6):1029-1034
The extracellular polysaccharides (EPS) of N. flagelliforme were purified by DEAE anion exchange chromatography and Sephadex G100 gel filtration chromatography. And two main components named NFPS1 and NFPS2 were obtained respectively. The physico-chemical characteristics of NFPS2 were analyzed and compared with NFPS0, which was obtained from field colony of N. flagelliforme. These results showed that both of NFPS2 and NFPSO were composed of four monosaccharides: glucose, xylose, galactose and mannose. The apparent molecular weight of NFPS2 and NFPS0 was estimated to be 2.79 x 10(5), 2.26 x 10(5) respectively. They are non-sulfated polysaccharides, free of protein and nuclear acid. The thermal analysis indicated that there was a decomposition peak at 245 degrees C in thermogravimetric (TG) curves. However, the microstructure analysis showed that they had different porous structures.
Extracellular Space
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chemistry
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Nostoc
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chemistry
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Polysaccharides
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chemistry
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isolation & purification
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Polysaccharides, Bacterial
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chemistry
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isolation & purification
4.Chemical synthesis of a synthetically useful L-galactosaminuronic acid building block.
Chun-Jun QIN ; Hong-Li HOU ; Mei-Ru DING ; Yi-Kuan QI ; Guang-Zong TIAN ; Xiao-Peng ZOU ; Jun-Jie FU ; Jing HU ; Jian YIN
Chinese Journal of Natural Medicines (English Ed.) 2022;20(5):387-392
Most bacterial cell surface glycans are structurally unique, and have been considered as ideal target molecules for the developments of detection and diagnosis techniques, as well as vaccines. Chemical synthesis has been a promising approach to prepare well-defined oligosaccharides, facilitating the structure-activity relationship exploration and biomedical applications of bacterial glycans. L-Galactosaminuronic acid is a rare sugar that has been only found in cell surface glycans of gram-negative bacteria. Here, an orthogonally protected L-galactosaminuronic acid building block was designed and chemically synthesized. A synthetic strategy based on glycal addition and TEMPO/BAIB-mediated C6 oxidation served well for the transformation of commercial L-galactose to the corresponding L-galactosaminuronic acid. Notably, the C6 oxidation of the allyl glycoside was more efficient than that of the selenoglycoside. In addition, a balance between the formation of allyl glycoside and the recovery of selenoglycoside was essential to improve efficiency of the NIS/TfOH-catalyzed allylation. This synthetically useful L-galactosaminuronic acid building block will provide a basis for the syntheses of complex bacterial glycans.
Carbohydrates
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Glycosides
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Oligosaccharides
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Oxidation-Reduction
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Polysaccharides/chemistry*
5.Acetylation of Rehmannia glutinosa polysaccharides and antioxidant activity of acetylated derivatives.
Jin LI ; Ting-Ting ZHANG ; Ding-Tao PU ; Ya-Jun SHI ; Zhen-Yu ZUO ; Chong-Ying LIU ; Yan CHEN ; Xiao-Bin JIA ; Peng ZHAO ; Liang FENG
China Journal of Chinese Materia Medica 2022;47(6):1539-1545
This study aims to acetylate Rehmannia glutinosa polysaccharides by acetic anhydride method, optimize process parameters and evaluate their antioxidant activity. With the degree of substitution(D_s) as a criterion, the effects of reaction time, acetic anhydride-to-polysaccharides ratio and temperature were investigated. Process parameters were optimized by single-factor experiment and response surface methodology. The infrared spectroscopy(IR) and scanning electron microscopy(SEM) proved the successful acetylation and were employed to preliminarily analyze the structural characteristics of acetylated derivatives. The results showed that the D_s was 0.327 under the optimal technological conditions, including m(acetic anhydride):m(R. glutinosa polysaccharides)=2.70, reaction time 3.0 h and temperature 48 ℃. Further, the antioxidant properties of acetylated derivatives were investigated in vitro and acetylation was found effective to improve the antioxidant activity of R. glutinosa polysaccharides. This study provides a reference for the further development and application of R. glutinosa polysaccharides.
Acetylation
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Antioxidants/pharmacology*
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Polysaccharides/pharmacology*
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Rehmannia/chemistry*
6.Study of mycelial polysaccharide from Paraisaria dubia of Ophiocordyceps gracilis asexual.
Yue WANG ; Zhao-Lan LI ; Fei-Ya SUO ; Dong-Ping SUN
China Journal of Chinese Materia Medica 2019;44(8):1704-1709
In order to provide a foundation for the development and application of Ophiocordyceps gracilis and increase the new resources of cordyceps,an asexual Paraisaria dubia was isolated from an O. gracilis fruit body. After 10 days of liquid fermentation,white globular mycelium and clear transparent fermentation were produced. The mycelium was extracted by hot water and precipitated with ethanol to obtain intracellular crude polysaccharide. The protein was deproteinized to obtain deproteinized polysaccharide. The intracellular pure polysaccharide was purified by Sepharose 4 B column chromatography and were analyzed by UV,IR,1 H-NMR,and13 CNMR data,as well as GC and HPLC. The results showed that the intracellular polysaccharide of P. dubia was composed of glucose,galactose and mannose with a molar ratio of 25. 54 ∶2 ∶1. It was a β-configuration glycosylic bond,containing pyranoside. The initial connection of polysaccharide was β(1→2)(1→4)(1→6) connection. This experiment provides a theoretical basis for the development and application of P. dubia.
Fungal Polysaccharides
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chemistry
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Galactose
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Glucose
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Hypocreales
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chemistry
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Mannose
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Mycelium
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chemistry
7.Isolation, purification and structure analysis of polysaccharides from Peacilomyces lilacinus.
Huai SHI ; Bo LIU ; Mingxing SU ; Sufang HUANG ; Yujing ZHU
Chinese Journal of Biotechnology 2010;26(8):1080-1087
The paper dealt with the characterization of polysaccharide of Paecilomyces lilacinus NH-PL-03 strain. First, we extracted and purified exude polysaccharide from the fungal fermentation broth by ethanol depositing method. Second, the proteins were removed by the Sevage method from the crude polysaccharide. Third, the purified polysaccharide (EP-1) was obtained after Superdex G-75 column separation. The results of UV-spectrometer and Sephacryl S-200 HR chromatography experiments showed that the EP-1 was a homogeneous pure polysaccharide with molecular weight of 35.2 kDa. Tested by paper chromatography analysis using the complete hydrolysis by sulfuric acid, we found that the EP-1 comprise single component as glucose. The chemical structure of EP-1 was confirmed as a kind of linear glucan linked by beta-(1,3) linkage. The Congo red reaction performed that EP-1 probable presented a triple-helical conformation in the dilute alkali.
Molecular Structure
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Paecilomyces
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chemistry
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Polysaccharides, Bacterial
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chemistry
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isolation & purification
8.Structures and bioactivity of polysaccharides from isatidis radix.
Liwei HE ; Xiang LI ; Honglan WANG ; Jianwei CHEN ; Dongsong SUN ; Mingyan WANG
China Journal of Chinese Materia Medica 2011;36(16):2179-2182
OBJECTIVETo investigated the chemical structures and bioactivity of polysaccharides from Isatidis Radix.
METHODPolysaccharides were extracted and purified by column chromatograph and their chemical structures were identified by UV, IR, NMR, periodic acid oxadation and Smith degradation method and their stimulation effects to macrophage were evaluated by using MTT method.
RESULTFive polysaccharides, polysaccharide A , B, C, D and E were gotten and their molecular weights were 2 000, 1 757.1, 1 34 2.7, 955.6, 11.7 kDa, respectively. Polysaccharide A was composed of arabinose, polysaccharide E was composed of arabinose and galactose, polysaccharides B, C, D were composed of glucose and 1 --> 2, 1 --> 3, 1 --> 4, 1 --> 6 linkages existed in polysaccharides A-E, of A, B, C, D, E were alpha-configurations. Polysaccharides B, C and D showed better bioactivity than polysaccharides A and E with stimulation index (SI) of 5.31, 4.76, 5.17.
CONCLUSIONFive polysaccharides are seperated firstly from Isatidis Radix.
Animals ; Isatis ; chemistry ; Magnetic Resonance Spectroscopy ; Mice ; Polysaccharides ; chemistry ; pharmacology
9.Study on mechanism and structure-activity relationship of hypolipidemic polysaccharides: a review.
Xia YANG ; Ying-Shu FENG ; Shan-Shan TONG ; Jiang-Nan YU ; Xi-Ming XU
China Journal of Chinese Materia Medica 2018;43(20):4011-4018
Hypolipidemic polysaccharides have notable activity and safety with a range of diverse sources. In this paper, the classification of hypolipidemic polysaccharides was carried out into polysaccharide sulfate, glycosaminoglycan, homopolysaccharide and heteropolysaccharide. The hypolipidemic activity mechanism and structure-activity relationship hypothesis of those polysaccharides in recent years were briefly reviewed therefore to provide references for the study and product development of polysaccharides.
Hypolipidemic Agents
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chemistry
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pharmacology
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Polysaccharides
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chemistry
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pharmacology
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Structure-Activity Relationship
10.Process optimization for extraction and purification of polysaccharides from Cistanche deserticola.
Xing-Hui XIAO ; Hai-Xia GAO ; Hong-Hui LI ; Xiao-Hui JI ; Xiao-Nan CUI ; Gui-Fang LI ; Yu-Xia LI
China Journal of Chinese Materia Medica 2019;44(3):475-481
In this study, taking Cistanche deserticola in Xinjiang as the experimental material, the optimal process for extracting polysaccharides from C. deserticola with water extraction was studied by using single factor and orthogonal experiment. Its effects on protein removal and polysaccharides retaining were investigated by using Sevag, enzymatic method or combination of these two methods, so as to determine the optimal method for protein removal from polysaccharides of C. deserticola; the decolorization and purification methods such as macroporous resin of AB-8 and activated Carbon were used to determine the optimal process. The results showed that the extraction rate of polysaccharides from C. deserticola was 18.40% during the optimal process of the water extraction as follows: extraction temperature 75 ℃, extraction time 165 min and solid-liquid ratio 1∶55. The protein removal rate can reach 31.40% and polysaccharide retention rate can reach 96.00% under the optimal protein removal process: temperature 50 ℃, time 2 h, and papain dosage 0.2%. The decolorization rate of activated Carbon and macroporous resin called AB-8 was 80.37% and 86.43%, and the recovery rate of polysaccharides was 77.05% and 91.93%, respectively, suggesting that macroporous resin was more suitable for decoloration. Macroporous resin named AB-8 increased the purity of the polysaccharide crude extract from 67.70% to 84.80% under the following conditions: concentration of the sample 4 g·L~(-1), concentration of the eluent 60% ethanol, and the flow rate 1 mL·min~(-1), showing significant purification effect.
Cistanche
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chemistry
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Plant Extracts
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chemistry
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Polysaccharides
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isolation & purification
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Temperature
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Water