银耳子实体多糖的分离、分析及生物活性

银耳(Tremella fuciformis Berk.)子实体经热水提取、去蛋白、柱层析分离纯化得银耳子实体多糖(简称TF)精品。TF总糖含量75.7%,其中含葡萄糖醛酸14.7%。TF由岩藻糖、阿拉伯糖、木糖、甘露糖、葡萄糖和葡萄糖醛酸组成,摩尔比为0:92:0.49:0.1 8:1.00:1.15:0.57,分子量115000。急性和亚急性毒性试验表明TF的毒性很小,小鼠腹腔注射,LD_(so)为96700±143.1 5mg/kg,狗静脉注射前后,血象、肝肾功能及组织切片均无病理性变化和损伤。TF有多种生物活性;可增加脾指数、半数溶血值和E玫瑰花结形成率,促进巨噬细胞吞噬功能、淋巴细胞转化和血清蛋白质的生物合成,并有明显抗放射、升高白血球、抗炎和红细胞凝集作用等。     

浒苔多糖的分离、纯化和分析

浒苔（Ｅｎｔｅｒｏｍｏｒｐｈａｐｒｏｌｉｆｅｒａ）经热水提取,Ｓｅｖａｇｅ法除去蛋白质,用乙醇沉淀,ＳｅｐｈａｄｅｘＧ－１００柱层析,得浒苔多糖（简称ＥＰ）精制品。经ＳｅｐｈａｄｅｘＧ－２００柱层析鉴定为单一对称性洗脱峰。红外光谱分析具有多糖特征吸收峰,紫外光谱分析未见有核酸和蛋白质的特征吸收峰。总糖含量为８８．８％,其中糖醛酸含量为３３．６％。单糖组成为Ｌ－阿拉伯糖、Ｌ－岩藻糖、Ｄ－甘露糖、Ｄ－半乳糖及Ｄ－葡萄糖,平均分子量为２５０００。     

长白楤木根、茎、叶水溶性多糖的纯化及组成分析

长白楤木是一种具有药用价值的植物。本实验用热水浸提、醇析、Severge法除蛋白,水相透析得到粗多糖。粗多糖经过DEAE-纤维素柱层析洗脱,再经SephadexG-200葡聚糖凝胶柱层析后,得到4种纯多糖,命名为根(gNaCl)茎(JH2O)茎(JNaCl)叶(YNaCl)。红外光谱和其完全水解的高效液相色谱分析表明,4种多糖不含硫酸基团和乙酰基,具备β-糖苷键以及糖类特征吸收峰。其中,根的NaCl洗脱多糖组分为D-半乳糖醛酸、D-果糖、D-葡萄糖,茎的水洗脱液多糖组分为D-甘露糖、D-葡萄糖,茎的NaCl洗脱多糖组分D-葡萄糖、D-半乳糖,叶的洗脱多糖组分为D-半乳糖醛酸和D-山梨糖。     

浒苔多糖的分离,纯化和分析

浒苔经热水提取,Ｓｅｖａｇｅ法除去蛋白质,用乙醇沉淀,Ｓｅｐｈａｄｅｘ－Ｇ－１００柱层析,得浒苔多糖（简称ＥＰ）精制品。经ＳｅｐｈａｄｅｘＧ－２００柱导析鉴定为单一对称性洗脱峰。红外光谱分析具有多糖特征吸收峰,紫外光谱分析表未见有核酸和蛋白质的特征及吸上峰,总糖含量为８８．８％,基中糖酸酸含量为３３．６％。单糖组成为Ｌ－阿拉伯糖,Ｌ－岩藻糖,Ｄ－甘露糖,Ｄ－半乳糖及Ｄ－葡萄糖。平均分子量为２５００     

仙草多糖的分离纯化及鉴定

仙草(Mesona Chiliensis Benth)是我国的一种民间草药。用0.5％NaHCO_3抽提,乙醇沉淀得到了仙草多糖的粗品。经过H_2O_2脱色处理,Sephadex G-75柱层析和硫酸—苯酚法收集单一峰部分,得到仙草多糖纯品(简称MCPS),薄层层析、凝胶电泳和醋纤薄膜电泳证明其为均一性多糖。红外光谱扫描表明,它具有典型的多糖吸收峰。HPLC法测得相对分子量为4.3×10~4,经薄层层析确定MCPS的单糖组成为葡萄糖、半乳糖、阿拉伯糖、木糖、鼠李糖,半乳糖醛酸及一种未知单糖。药理实验表明,它具有免疫促进作用与抑瘤效应。     

长松萝多糖的研究

长松萝经三氯甲烷抽提后风干,用热水提取,乙醇沉淀,经微晶纤维素柱层析纯化,得白色粉末状多糖USL。USL经Sephadex G-150柱层析证明为一组均匀多糖,其糖的含量为89.52％。经气相色谱检定,由阿拉伯糖(Ara)、木糖(Xy1)、甘露糖(Man)、葡萄糖(Glc)聚合而成,其克分子比为0.31:0.05:1.00:18.10。经Sephadex G-200柱层析测定,平均分子量为30×10~4,经高碘酸氧化,Smith降解,有甲酸、丙三醇、赤藓醇生成。红外光谱在896cm-~1处有吸收,证明USL多糖结构主要以β(1→4)、β(1→6)甙键连接而成的杂多糖。     

根瘤菌TISTR 386胞外酸性多糖的结构研究

根瘤菌TISTR 386胞外酸性多糖有二种九糖的重复单位构成。重复单位主要成份是D一葡萄糖,D一半乳糖和D一葡萄糖醛酸,它们的克分子比例分别是6：l：2和5：2：2。另外还含有一些丙酮酸和醋酸。甲基化分析表明,这个多糖由一个(1→3)键,一个(1→6)键,三个(1→4)键,一个(1→4,1→6)键连结的葡萄糖残基,一个(1→3)键连结的D-半乳糖残基,以及一个(1→3)键,一个(1→4)键连结的D-葡萄糖醛酸残基所组成。非还原末端糖残基是D葡萄糖或是带有丙酮酸的D一半乳糖,这也是二种九糖重复单位区别所在。     

不同提取方法对软枣猕猴桃多糖单糖组成及抗氧化活性的影响

采用高温水提工艺、低温水提工艺和微波辅助工艺从软枣猕猴桃中提取得到三种多糖,依次命名为AAP-1、AAP-2和AAP-3,对三种多糖的理化性质、单糖组成和抗氧化活性进行了研究.理化性质鉴定结果表明:三种多糖均不含酚羟基及还原糖,都含有一定量的糖醛酸和蛋白质,AAP-1含有淀粉,AAP-2和AAP-3不含淀粉;单糖组成结果表明:三种多糖均由甘露糖、鼠李糖、葡萄糖醛酸、半乳糖醛酸、葡萄糖、半乳糖、阿拉伯糖7种单糖组成,其中,AAP-1葡萄糖的摩尔百分含量最高,为94.72％;AAP-2半乳糖和阿拉伯糖的摩尔百分含量较高,分别为24.75％、38.37％;AAP-3半乳糖醛酸的摩尔百分含量较高,为16.05％.抗氧化实验结果表明:AAP-1抗氧化活性最弱;AAP-3抗氧化活性最强,其清除DPPH自由基和羟基自由基的IC50分别为1.2、2.7 mg/mL.     

金耳子实体和发酵菌丝体多糖的分离、纯化与结构的比较研究

人工段木栽培的金耳（TremellaaurantialbaBandonietZang）子实体和深层培养的金耳菌丝体分别经沸水提取、醇析、透析,Sevape法脱蛋白、SephadexG—100柱层析纯化,得子实体纯多糖TAf和菌丝体纯多糖TAm。玻璃纤维纸电泳表明TAm和TAf为单一均匀多糖。薄层层析表明,TAm的单糖组成为半乳糖、葡萄糖、甘露糖、岩藻糖、鼠李糖。TAf的单糖组成为葡萄糖、甘露糖、木糖、鼠李糖、葡萄糖醛酸。UV分析表明组成中不含核酸和蛋白质。TAf经乙醇分级分离得TAf－1和TAf－2。TAf－1经SephadexG－100柱层析表明其为单一均匀物质。红外光谱分析证明,TAf－1和TAf－2有多糖吸收峰,存在吡喃环,α-糖苷键和β-糖苷键连接。     

白芍多糖分离纯化及性质分析

采用热水浸提.乙醇沉淀法中药白芍(Paeonia Albiflora Pall)水提取液中分离纯化得到一种多糖蛋白复合物,命名为PAⅡ(Paeonia Albiflora Pall PolysaccharideⅡ,PAⅡ).电泳、FPLC和HPLC检测其纯度;红外光谱和气相色谱法对其结构和组成进行初步分析.结果表明PAⅡ为均一多糖组分,是以D-葡萄吡喃糖为主的多糖-蛋白复合物,总糖含量为85.0%,蛋白含量为13.2%;该糖为吡喃型结构,糖肽键为非O-型;单糖组分为葡萄糖、阿拉伯糖、甘露糖、鼠李糖、木糖,摩尔比为153:2.6:1.25:1:1,平均分子量为5.25×104.     

红栓菌多糖的分离纯化与分析

液体发酵得到的红栓菌（Ｐｙｃｎｏｐｏｒｕｓｃｉｎｎａｂａｒｉｕｓ）菌丝经沸水抽提,胰匀浆处理和Ｓｅｖａｇｅ法除蛋白,ＤＥＡＥＳｅｐｈａｄｅｘＡ－２５与ＳｅｐｈａｄｅｘＧ－１５０柱层析,乙醇沉淀得到二种多糖组分（简称ＰＣ＿１、ＰＣ＿２）。经聚丙烯酰胺电泳,冻融分析鉴定为均一体。ＰＣ＿１和ＰＣ＿２的,总糖含量分别为８１．５％和７７．３％,分子量分别为２３０００和１３０００．两者的红处光谱具多糖特征吸收峰,在紫外区无明显吸收现象,基本不含氮。ＰＣ＿１单糖组成为Ｄ－葡萄糖、Ｄ－半乳糖及Ｄ－甘露糖,ＰＣ＿２为Ｄ－半乳糖、Ｄ－甘露糖。     

雪灵芝多糖的分离纯化及免疫活性评价

为分离纯化雪灵芝(Arenaria kansuensis)多糖,并对纯化组分进行分子量测定、单糖组分分析及免疫活性评价。实验采用水提醇沉法提取雪灵芝粗多糖(Arenaria kansuensis crude polysaccharide, AKCP);以DEAE-52纤维素柱对AKCP进行分离纯化,获得5个雪灵芝多糖组分AKP-1～AKP-5,进一步采用葡聚糖凝胶G-75柱对AKP-2进行分离纯化获得AKP-2a多糖组分。苯酚-硫酸法测定AKCP、AKP-2及AKP-2a的总糖含量分别为52%、70%和79%;凝胶渗透色谱-十八角度激光光散射(GPC-MALS)法检测AKP-2a的重均分子量Mw为2.07×10~5Da、数均分子量Mn为9.838×10~4Da;HPLC法检测AKP-2a是由半乳糖醛酸、甘露糖、核糖、鼠李糖、葡萄糖醛酸、葡萄糖、半乳糖、木糖、阿拉伯糖、岩藻糖10种单糖组成,其摩尔比为1∶0.25∶0.01∶0.20∶0.11∶0.25∶0.61∶0.07∶0.21∶0.12;以MTT法检测体外培养小鼠脾淋巴细胞增殖,AKCP、AKP-2及AKP-2a各浓度组SI水平,均明显高于对照组(P<0.05)。经NO释放实验及IFN-γELISA检测,AKP-2a各浓度组小鼠腹腔巨噬细胞培养上清中二者的水平,较对照组呈浓度依赖性增高(P<0.01)。综上结果,本研究通过分离纯化,获得了总糖含量较高的雪灵芝多糖AKP-2a组分,初步确定其分子量范围及单糖组成,并证实其具有激活淋巴细胞增殖、促进巨噬细胞功能的生物活性。     

Structure of extracellular polysaccharides of Escherichia coli strains 36M, 72M, and 29M isolated from coligranuloma of chick intestine. I. Polysaccharide from E. coli 36M.

An extracellular polysaccharide was isolated from culture broth of Escherichia coli 36M, and fractionated on a column of Sephadex G-150 into two fractions; the high molecular weight portion (85% of the total polysaccharide) contained pyruvic acid, and showed a positive immune reaction with anti-Ps-I-serum obtained from a rabbit. The low molecular weight portion (15% of the total polysaccharide) showed a negative immune reaction. The methylation, Smith's degradation, partial acid hydrolysis and methanolysis of the higher molecular weight polysaccharide revealed a repeating structure as follows: (see article).     

Structure of shigella antigens. Heterogeneity of specific polysaccharides of 2 Shigella flexneri strains and 2 Sh. flexneri/Escherichia coli hybrids]

The S-specific polysaccharide from 2 Sh. flexneri wild strains (with serological var. X- and var. Y-specificity, respectively) and 2 Sh. flexneri E. coli hybrids (with the same specificities) can be separated by means of gel chromatography on Sephadex G-200 and G-50 into altogether 6 fractions per strain. Fraction G-200/1 (molecular weight greater than 10(6)D) represents a polymer consisting nearly exclusively of glucose and is present mainly in the two Y-type strains, much less in the two X-type strains. Fractions G-200/2 and G-200/3 (molecular weight approximately 10(5)D and approximately 2 - 10(4)D, respectively) seem to consist mainly of the S-specific side chains while fraction G-50/2 (molecular weight approximately 2000 D) presumably contains an SR-polysaccharide (core with one repeating unit.) Fraction G-50/3 (molecular weight approximately 100 D) contains the core polysaccharide and fraction G-50/4 splitting products (mainly KDO). No significant differences in chromatographical behaviour and quantitative composition could be found between the polysaccharides of the wild strains and the hybrid strains. Because of the well-known stability of the glucosaminyl linkages the sugar analysis was not only performed after acidic hydrolysis. In some cases the acid hydrolysate was reacted with HNO2 to cleave the glucosaminyl linkages. In most cases the values obtaines now were higher than those obtained directly.     

Extraction, purification and antibacterial activities of a polysaccharide from spent mushroom substrate

To contribute towards effective exploitation and utilization of spent mushroom substrate (SMS), a water-soluble polysaccharide named PL was isolated and purified from SMS. The total sugar content and monosaccharide composition were analyzed by phenol-sulfuric acid method and capillary electrophoresis, and infrared spectroscopy was also performed for structure characterization. The results showed that the total sugar content of crude polysaccharide from SMS was about 25.8%, the polysaccharide contained two fractions (PL1 and PL2), which was mainly composed of glucose, rhamnose and mannose with a molar ratio of 1:3.13:1.16. The attributions of the main absorptions of both PL1 and PL2 were characteristic of glycosidic structures, and the FT-IR spectra of PL2 and lentinan were very similar. Escherichia coli, Staphylococcus aureus and Sarcina lutea were used to study the antibacterial activity and minimal inhibitory concentrations (MICs) of the polysaccharide. The antibacterial activity of polysaccharide from SMS against E. coli was the strongest, while the weakest against S. lutea, and the MICs of PL2 were 12.5, 25 and 100 μg/mL, respectively.     

The structure of the biological repeating unit of the O-antigen of Hafnia alvei O39.

On mild acid-catalysed degradation of the lipopolysaccharide from Hafnia alvei O39 followed by gel filtration of Sephadex G-50, the O-specific polysaccharide and three oligosaccharides were obtained, which represent the core substituted with 0-2 O-antigen repeating-units. On the basis of sugar and methylation analyses, 13C-n.m.r. data, solvolysis of the polysaccharide with anhydrous hydrogen fluoride, and computer-assisted 13C-n.m.r. analysis of the Smith-degraded polysaccharide, it was concluded that the biological repeating unit of the O39 antigen was Formula; see text     

淫羊藿多糖的分离纯化及结构初步分析

从淫羊藿中提取多糖并鉴定其初步结构和单糖组成.采用超声-水提醇沉法提取粗多糖、Sevag法去蛋白、DEAE-52纤维素及Sephadex G-100柱层析法纯化得到淫羊藿多糖EPSⅠ-1和EPSⅡ-1.应用紫外光谱法和红外光谱法对其结构做初步分析.采用高效液相色谱法(HPLC)测定其单糖组成及摩尔比.均一的EPSⅠ-1和EPSⅡ-1多糖在紫外和红外中具有多糖的特征吸收峰,组成中含有吡喃环结构;EPSⅠ-1的单糖组成为鼠李糖和葡萄糖,摩尔比为1:1.13;EPSⅡ-1的单糖组成为果糖、葡萄糖和一个不确定的糖,摩尔比为1:1.91.有效地分离纯化了淫羊藿多糖,这为淫羊藿多糖的广泛应用奠定了实验基础.     

塔拉种子多糖脱蛋白的正交优化及其抗氧化性研究

以塔拉（Caesalpinia spinosa）种子为原料,研究了塔拉种子多糖的脱蛋白工艺及塔拉多糖的抗氧化性质。以多糖损失率和蛋白脱除率为评价指标,比较Sevage法、三氯乙酸法和木瓜蛋白酶法对塔拉多糖的脱蛋白效果。利用正交优化组合实验设计原理,采用四因素三水平的正交分析法,对木瓜蛋白酶法脱蛋白进行正交优化。结果表明：塔拉多糖最佳脱蛋白工艺条件为酶添加量0.15mL、酶解时间90min、酶解温度60℃、酶解pH=6,蛋白脱除率95.19%,多糖保留率75.02%。通过对塔拉多糖抗氧化性的研究,发现塔拉多糖总抗氧化性较好,对DPPH自由基有较强的清除作用。     

猴头菌丝多糖降血糖作用研究

目的:研究猴头菌丝多糖的降血糖作用。方法:以液体发酵生产的猴头菌丝体为原料,经热水浸提、浓缩、酒精沉淀获得菌丝粗多糖;以常规降糖药物格列本脲为阳性治疗对照,通过四氧嘧啶诱发小鼠糖尿病的预防试验,比较猴头菌丝多糖各剂量与格列本脲的降血糖效果。结果:猴头菌丝多糖得率为7.14%,粗多糖再经Sevage法去除蛋白质,获得猴头菌丝精多糖(HMP),得率为10.92%;猴头多糖高、中、低三个剂量均能有效的对抗四氧嘧啶诱发的高血糖;其中,高剂量的降血糖作用与格列本脲相比,差异极显著。结论:猴头菌丝多糖对四氧嘧啶型高血糖模型小鼠有降血糖作用,作用效果优于格列本脲,对糖尿病小鼠的胰腺具有一定的保护作用。