苦豆子多糖的分离纯化及初步结构北大核心CSCD

本文研究了苦豆子多糖的分离纯化以及初步结构。采用水溶醇沉方法提取粗多糖,然后通过离子交换层析分离纯化,采用高效凝胶渗透色谱、气相色谱、紫外吸收以及红外吸收光谱对多糖组分进行分析研究。获得苦豆子多糖相对分子质量均一性组分SPN和SPA,测得其平均相对分子质量分别为1.217×106Da和4.412×105Da,二者均不含蛋白质,且都具有红外的多糖特征吸收峰,其单糖组成(质量比)分别为阿拉伯糖∶木糖∶甘露糖∶葡萄糖∶半乳糖=7.90∶11.24∶86.24∶18.81∶16.89、鼠李糖∶阿拉伯糖∶木糖∶甘露糖∶葡萄糖∶半乳糖=1.50∶5.80∶1.72∶20.10∶1.64∶27.20。本文为苦豆子多糖的进一步研究开发提供理论依据。     

苦豆子多糖的分离纯化及初步结构

本文研究了苦豆子多糖的分离纯化以及初步结构。采用水溶醇沉方法提取粗多糖,然后通过离子交换层析分离纯化,采用高效凝胶渗透色谱、气相色谱、紫外吸收以及红外吸收光谱对多糖组分进行分析研究。获得苦豆子多糖相对分子质量均一性组分SPN和SPA,测得其平均相对分子质量分别为1.217×106Da和4.412×105Da,二者均不含蛋白质,且都具有红外的多糖特征吸收峰,其单糖组成(质量比)分别为阿拉伯糖∶木糖∶甘露糖∶葡萄糖∶半乳糖=7.90∶11.24∶86.24∶18.81∶16.89、鼠李糖∶阿拉伯糖∶木糖∶甘露糖∶葡萄糖∶半乳糖=1.50∶5.80∶1.72∶20.10∶1.64∶27.20。本文为苦豆子多糖的进一步研究开发提供理论依据。     

香菇子实体蛋白我糖的分离纯组成结构分析

香菇〔Lentinus edodes（Berk.）Sing.〕子实体经热水提取,乙醇沉淀,得多糖粗品（Le）。Le脱游离蛋白,脱色,经DEAE－纤维素柱及纤维素凝胶柱层析分离酏化和到Le－1、Le－2和 Le－33个级分。经聚丙烯酰胺凝胶电泳和SepharoseCL－6B柱层析鉴定为分子量分布均一的蛋白多糖。凝胶渗透色谱法测得Le－1、Le－2和 Le－3分子量分别为954000,90000和14000。3个级分均由阿拉伯糖、木糖、甘露糖、半乳糖、葡萄糖和葡萄糖醛酸组成,气相色谱测得Le－1的阿拉伯糖、木糖、甘露糖、半乳糖、葡萄糖的摩尔比为0．39：0．46：1．00：0．93：14．13;Le－2为0．19：0．41：1．00：0．93：10．72;Le－3为0．31：0．47：1．00：1．15：8．92。Le－1、Le－2和 Le－3葡萄糖 醛酸含量（％）分别为24．10、34．77和40．05,蛋白质含量（％）分别为2．01、7．28和25．31,红外扫描确定Le－1和Le－2 的糖苷键为a型,Le－3为β型。     

丹皮多糖PSM2b的纯化及其理化性质研究

从中药丹皮(MoutanCortex)蒸馏水浸提液得到粗多糖,经DEAE Cellulose 52柱层析分离得到降血糖组分PSM2b,Surperdex200柱层析进一步纯化得到PSM2b A和PSM2b B两个组分,快速层析纯化系统(FPLC)和电泳鉴定为均一的多糖蛋白复合物。FPLC法测定A和B两组分的分子量分别为1.16×105、1.30×104,苯酚 硫酸法测得其总糖含量分别为76.91%、32.00%,Folin 酚法测得其蛋白含量分别为9.90%、42.60%。气相色谱分析PSM2b A的单糖组成为:L 鼠李糖、L 岩藻糖、L 阿拉伯糖、D 木糖、D 甘露糖、D 葡萄糖、D 半乳糖,摩尔比依次为1.00∶0.18∶4.30∶0.50∶1.30∶2.41∶6.97,PSM2b B的单糖组成为:L 鼠李糖、L 岩藻糖、L 阿拉伯糖、D 葡萄糖、D 半乳糖,摩尔比依次为1.00∶1.17∶0.183∶2.34∶4.18。两者的红外光谱呈现多糖吸收特征峰,含有吡喃糖苷键。β消去反应初步证明所提取的两种糖蛋白不存在O 型糖肽键。     

柿子多糖的分离纯化和结构分析

利用水提醇沉提取柿子多糖(WPP),经季铵盐沉淀法和凝胶柱层析对柿子粗多糖进行分离纯化,得到了水溶性的柿子粗多糖(WPP1)和盐溶性的柿子粗多糖(WPP2)两个柿子多糖组分。通过对理化性质、分子量和单糖组成测定结果分析,确定WPP1是含有α糖苷键的化合物,由L-鼠李糖、L-阿拉伯糖、D-葡萄糖和D-半乳糖四种单糖组成,四种单糖的摩尔比为0.8831∶0.6862∶0.7022∶1,分子量为2.05×105Da。WPP2由L-阿拉伯糖和D-半乳糖两种单糖组成,其摩尔比为0.8466∶1,分子量为2.63×105Da。WPP1和WPP2均表现出吡喃糖的特征吸收。     

香菇菌丝体多糖LeBDl—1的分离纯化和分析

香菇(Lentinula edodes)465菌株用添加啤酒发酵醪泥的培养基发酵培养10d后,菌丝体经洗涤、热水浸提、乙醇沉淀,再经DEAE-纤维素柱(CI—型)分离和Sephadex G—100柱层析纯化,得到白色絮状多糖LeBD1—1。经醋酸纤维薄膜电泳和HPLC检测纯度,LeBD1—1为均一成分。HPLC法测得分子量为130000Da,红外光谱测定其糖苷键为α型。经完全酸水解后用硅胶薄层层析和气相色谱法分析单糖组成,LeBD1—1中含D-甘露糖、D-葡萄糖、D-半乳糖、D-木糖和D-阿拉伯糖,各单糖摩尔比为2.45:1.00:0.03:0.08:0.10,是一种以含甘露糖为主的杂多糖。     

凉粉草胶的初级结构与流变性质的研究

凉粉草胶经过DEAE-Sepharose FF分离可以得到中性糖和酸性糖两种组分,其中中性糖的尖峰分子量Mp为5227,酸性糖的尖峰分子量为6566,前者的单糖组成(以摩尔百分比计)为半乳糖:葡萄糖:甘露糖:木糖:阿拉伯糖:鼠李糖=9.9∶15.3∶4.31∶1.48∶11.6∶1,后者的单糖组成为半乳糖:葡萄糖:甘露糖:木糖:阿拉伯糖:半乳糖醛酸:鼠李糖=2.66∶1∶0.37∶2.29∶12.5∶23.5∶5.99。两者再经Sephadex G-100凝胶色谱分离,均为单一对称峰,酸性糖经琼脂糖凝胶电泳,甲苯胺蓝显色为均一斑点,表明所分离得到的两种组分都为均一组分。紫外光谱分析表明两种糖都不含蛋白质或肽段,红外光谱分析表明NMBG和AMBG都具有多糖特征吸收峰,NMBG仅在872 cm-1附近有吸收峰,表明其结构中只有β-糖苷键,AMBG在896和858 cm-1附近有吸收峰,表明其结构中既有β-糖苷键,又有α-糖苷键。流变学实验表明在1%~5%(w/w)内中性糖不具有流变学性质,但酸性糖却表现出明显地流变学性质。     

猴头菌子实体和菌丝体多糖的分离纯化与理化特征的比较*

人工栽培的猴头菌子实体和固体培养的菌丝体分别经热水提取,浓缩、醇析后得子实体粗多糖（HFP）和菌丝体粗多糖（HMP）,HFP的得率和多糖含量均高于HMP;两者再经透析、Sevag法脱蛋白、乙醇分级沉淀、SephadexG-100柱层析纯化,得子实体纯多糖hfp-1和菌丝体纯多糖 hmp-2,经HPLC检测hfp-1和hmp-2为单一均匀多糖。气相色谱分析表明hfp-1的单糖组成为阿拉伯糖、甘露糖、半乳糖和葡萄糖,摩尔比为：0.12：0.04：1.00：0.71;hmp-2的单糖组成为阿拉伯糖、木糖、甘露糖、半乳糖和葡萄糖,摩尔比为：0.25：0.41：0.31：1.00：0.29。紫外光谱分析表明组成中不含核酸和蛋白质,红外光谱分析二者均有多糖特征吸收峰,hfp-1有β-型连接的吸收峰,hmp-2无明显的β-型连接的吸收峰;经HPLC进行分子量测定,hfp-1的分子量为54000,hmp-2的分子量为68000。猴头菌子实体多糖和菌丝体多糖在化学组成上有一定的差异。     

猴头菌子实体和菌丝体多糖的分离纯化与理化特征的比较

人工栽培的猴头菌子实体和固体培养的菌丝体分别经热水提取,浓缩、醇析后得子实体粗多糖（HFP）和菌丝体粗多糖（HMP）,HFP的得率和多糖含量均高于HMP;两者再经透析、Sevag法脱蛋白、乙醇分级沉淀、SephadexG-100柱层析纯化,得子实体纯多糖hfp-1和菌丝体纯多糖hmp-2,经HPLC检测hfp-1和hmp-2为单一均匀多糖,气相色谱分析表明hfp-1的单糖组成为阿拉伯糖、甘露糖、半乳糖和葡萄糖,摩尔比为：0.12:0.04:1.00:0.71;hmp-2的单糖组成为阿拉伯糖、木糖、甘露糖、半乳糖和葡萄糖,磨尔比为：0.25:0.41:0.31:1.00:0.29。紫外光谱分析表明组成中不含核酸和蛋白质,红外光谱分析二者均有多糖特征吸收峰,hfp-1有β-型连接的吸收峰,hmp-2无明显的β-型连接的吸收峰;经HPLC进行分子量测定,hfp-1的分子量为54000,hmp-2的分子量为68000,猴头菌子实体多糖和菌丝体多糖在化学组成上有一定的差异。     

画眉草多糖提取及其保湿性能研究

本文以脱脂画眉草为原料,用纤维素酶超声辅助法提取多糖;经脱蛋白、脱色、DEAE-纤维素分离纯化,得到3种多糖TPS1、TPS2、TPS3,提取率分别为0.42％、0.66％、0.54％.通过高效凝胶渗透色谱(HPGPC)分析,3种多糖的分子量分别为7886 Da、8467 Da、6366 Da;高效离子色谱(HPIC)测得TPS1组成单糖为果糖,TPS2、TPS3组成单糖为阿拉伯糖、半乳糖、葡萄糖、木糖、甘露糖、果糖、核糖和一种未知糖.对多糖TPS2的保湿和吸湿性研究结果表明TPS2的保湿和吸湿能力均优于常用保湿剂甘油、聚乙二醇400、1,3-丁二醇、壳聚糖.     

苦瓜多糖的提取、分离纯化及组成性质

正交实验确定提取工艺后,用热水提取法得到苦瓜多糖(MCP).对MCP进行DEAE-32离子交换层析分离,得到3个多糖组分MCP1、MCP2和MCP3. 进一步采用Sephacryl S-400凝胶层析进行分离,经凝胶层析和高效液相色谱检测表明,MCP1、MCP2为均一性多糖组分.通过高效液相凝胶色谱法测定了两者的相对分子质量分别为1.16×106和7.45×105.用PMP衍生化法测定其单糖,结果表明: MCP1系由Man、Rham、GlcUA、GalUA、Glu、Gal、Xyl、Ara等单糖组成的杂多糖,摩尔比为1.03:2.93:1.00:14.95:2.16:30.70:2.85:4.50.MCP2系由Rham、GalUA、Gal、Xyl、Ara等单糖组成的杂多糖,对应的摩尔比为1.63:21.88:4.66:1.00:1.29.紫外光谱表明该多糖不含蛋白质和核酸.     

枸杞多糖的提取纯化及组成分析

采用水提法从枸杞中提取分离枸杞多糖(LBP)用DEAE纤维素柱色谱和凝胶柱色谱进行分离纯化,采用GPC-LLS法、红外光谱和气相色谱等方法对其组成进行研究。结果表明LBP含有3个级分,LBP的分子质量约为1.497×105,由阿拉伯糖(Ara),鼠李糖(Rha),木糖(Xyl),甘露糖(Man),半乳糖(Gal)和葡萄糖(Glc)6种中性单糖组成。     

Characterisation of extracellular polysaccharides from suspension cultures of apple (Malus domestica)

The polymers secreted by suspension-cultured apple cells were composed of 85% carbohydrate (76% neutral sugar and 9% uronic acid) and 15% w/w protein. The extracellular polysaccharides (ECPs) contain 23% XG and 59% AGPs. The monosaccharide composition of the ECPs consisted of Gal, Ara, Glc and Xyl, with smaller amounts of Rha, Fuc and Man. Fractionation of the ECPs by anion-exchange chromatography yielded an unbound neutral fraction and a bound acidic fraction. Monosaccharide and linkage compositions of each fraction were determined. The neutral fraction (48% recovered carbohydrate) was composed of xyloglucan (XG; >90 mol%) which was purified by selective precipitation with Fehling’s solution to yield pure XG. The purified XG had a Glc:Xyl:Gal:Fuc ratio of 4.0:2.5:0.8:0.5; the XG was not O-acetylated. The structure of the secreted XG was similar to that extracted from apple-pomace. The acidic fraction (52% recovered carbohydrate) was composed primarily of arabinogalactan-proteins (AGPs) as detected by the β-glucosyl Yariv diffusion test. The AGP had a Gal:Ara ratio of 1.3: 1.0. Minor amounts of arabinan, xylan and mannan were also detected in the ECPs. This study is the first examination of the polysaccharides secreted by apple cells grown in suspension culture.     

Primary structure and configuration of tea polysaccharide

Polysaccharide is a class of natural macromole-cules of which many species have been found to carry significant biological activities. Although the research on activities of saccharide has been at a lower level in the past comparing to those of proteins and nucleic acids, much progress has been made in recent years because of accelerated activities worldwide[1]. Such progress has been made mostly in areas of structural analysis, and researches on structure-activity relation-ships. The biologic…     

草苁蓉根、茎水溶性多糖BRT的结构特征

本文以长白山区珍贵野生药用植物草苁蓉为研究对象 .草苁蓉又名“不老草” ,具有滋补强壮、益寿延年之功及补肾壮阳、润肠止血之效 ,为国家三级重点保护植物[1] .近年的研究发现 ,草苁蓉醇提物不仅可以清除体内的游离基 ,而且还可以显著增强机体的免疫能力 ,同时对草苁蓉化学成分的研究也在逐步深入[2 ] ,但对于草苁蓉多糖的系统研究尚未见报道 .为了更全面地认识和利用草苁蓉这一珍贵的植物资源 ,同时也为探讨多糖的结构与功能的关系 ,本文对草苁蓉根、茎的水溶性多糖ＢＲＴ组分进行了结构测定方面的研究 .1　材料和方法1 1　材料为本研究…     

The oligosaccharides of the glycoprotein pheromone of Volvox carteri f. nagariensis Iyengar (Chlorophyceae)

The sexuality-inducing glycoprotein of Volvox carteri f. nagariensis was purified from supernatants of disintegrated sperm packets of the male strain IPS-22 and separated by reverse-phase HPLC into several isoforms which differ in the degree of O-glycosylation. Total chemical deglycosylation with trifluoromethanesulphonic acid yields the biologically inactive core protein of 22.5 kDa. This core protein possesses three putative binding sites for N-glycans which are clustered in the middle of the polypeptide chain. The N-glycosidically bound oligosaccharides were obtained by glycopeptidase F digestion and were shown by a combination of exoglycosidase digestion, gaschromatographic sugar analysis and two-dimensional HPLC separation to possess the following definite structures: (A) Man beta 1-4GlcNAc beta 1-4GlcNAc; (B) (Man alpha)3 Man beta 1-4GlcNAc beta 1-4GlcNAc Xyl beta; (C) (Man alpha)2 Man beta 1-4GlcNAc beta 1-4GlcNAc; (D) (Man)2Xyl(GlcNAc)2. Xyl beta Two of the three N-glycosidic binding sites carry one B and one D glycan. The A and C glycans are shared by the third N-glycosylation site. The O-glycosidic sugars, which make up 50% of the total carbohydrate, are short (up to three sugar residues) chains composed of Ara, Gal and Xyl and are exclusively bound to Thr residues.     

Primary structure and configuration of tea polysaccharide

The monosaccharide composition of a tea polysaccharide (TGC) was determined by GC-MS method. Furthermore, the primary structure of tea polysaccharide and its configuration in the aqueous solution were investigated utilizing a combination of classical chemical methods and modern instrumental techniques including GC-MS, Proton NMR, UV and CD. The results indicate that TGC consists of 6 monosaccharides: Rha, Ara, Xyl, Glu, Man and Gal. The configuration of TGC in water solution is proposed to be an ordered helix. The possible primary structure of TGC was outlined as below: the basic structure of the main chain consists of Rha, Glu and Gal units. All three monosaccharides can potentially be connected to branch chains consisting of mainly Ara, and the linkages could be in β1 → 2, β 1 → 3, β 2 → 3 forms. When branch chain is absent in the basic structure of the main chain the linkage consists of only β 1 → 3; Xyl exists at the terminal end of either the main chain or the branch chain with β 1 → linkage.     

N-linked oligosaccharides of glycoproteins from Ginkgo biloba pollen, an allergenic pollen.

The pollen of Ginkgo biloba is one of the allergens that cause pollen allergy symptoms. The plant complex type N-glycans bearing beta1-2 xylose and/or alpha1-3 fucose residue(s) linked to glycoallergens have been considered to be critical epitopes in various immune reactions. In this report, the structures of N-glycans of total glycoproteins prepared from Ginkgo biloba pollens were analyzed to confirm whether such plant complex type N-glycans occur in the pollen glycoproteins. The glycoproteins were extracted by SDS-Tris buffer. N-Glycans liberated from the pollen glycoprotein mixture by hydrazinolysis were labeled with 2-aminopyridine and the resulting pyridylaminated (PA-)N-glycans were purified by a combination of size-fractionation HPLC and reversed-phase HPLC. The structures of the PA-sugar chains were analyzed by a combination of two-dimensional sugar chain mapping, IS-MS, and MS/MS. The plant complex type structures (GlcNAc2Man3Xyl1Fuc1GlcNAc2 (31%), GlcNAc2Man3Xyl1GlcNAc2 (5%), Man3Xyl1Fuc1GlcNAc2 (13%), GlcNAc1Man3Xyl1Fuc1GlcNAc2 (8%), and GlcNAc1Man3Xyl1GlcNAc2 (17%)) have been found among the N-glycans of the glycoproteins of Ginkgo biloba pollen, which might be candidates for the epitopes involved in Ginkgo pollen allergy. The remaining 26% of the total pollen N-glycans have the typical high-mannose type structures: Man8GlcNAc2 (11%) and Man6GlcNAc2 (15%).     

银耳多糖单糖组成分析的三种色谱方法比较

为选择一种准确快捷的方法测定银耳多糖的单糖组成,对薄层色谱法(TLC)、气相色谱法(GC)、高效液相色谱法(HPLC)三种色谱方法进行比较。结果表明,前两种方法的测定结果均不理想,而HPLC法,操作简便,灵敏度高,分离效果好,信息完整。测定结果为由葡萄糖、甘露糖、葡萄糖醛酸、木糖、岩藻糖组成,其摩尔比为0.24∶1.00∶0.06∶0.29∶0.25。HPLC法对酸性杂多糖组成糖分析是一种比较理想的选择。     

Structural features of N-glycans linked to glycoproteins from oil palm pollen, an allergenic pollen*

The pollen of oil palm (Elaeis guineensis Jacq.) is a strong allergen and causes severe pollinosis in Malaysia and Singapore. In the previous study (Biosci. Biotechnol. Biochem., 64, 820-827 (2002)), from the oil palm pollens, we purified an antigenic glycoprotein (Ela g Bd 31 K), which is recognized by IgE from palm pollinosis patients. In this report, we describe the structural analysis of sugar chains linked to palm pollen glycoproteins to confirm the ubiquitous occurrence of antigenic N-glycans in the allergenic pollen. N-Glycans liberated from the pollen glycoprotein mixture by hydrazinolysis were labeled with 2-aminopyridine followed by purification with a combination of size-fractionation HPLC and reversed-phase HPLC. The structures of the PA-sugar chains were analyzed by a combination of two-dimensional sugar chain mapping, electrospray ionization mass spectrometry (ESI-MS), and tandem MS analysis, as well as exoglycosidase digestions. The antigenic N-glycan bearing alpha1-3 fucose and/or beta1-2 xylose residues accounts for 36.9% of total N-glycans: GlcNAc2Man3Xyl1Fuc1GlcNAc2 (24.6%), GlcNAc2Man3Xyl1GlcNAc2 (4.4%), Man3Xyl1Fuc1-GlcNAc2 (1.1%), GlcNAc1Man3Xyl1Fuc1GlcNAc2 (5.6%), and GlcNAc1Man3Xyl1GlcNAc2 (1.2%). The remaining 63.1% of the total N-glycans belong to the high-mannose type structure: Man9GlcNAc2 (5.8%), Man8GlcNAc2 (32.1%), Man7GlcNAc2 (19.9%), Man6GlcNAc2 (5.3%).