银耳孢子多糖TF-A、TF-B、TF-C的分离、纯化及组成单糖的鉴定

用固体培养法获得的中国福建产银耳孢子(Tremella fucifromis Berk)经热水提取,三氯醋酸-正丁醇除杂蛋白、透析、乙醇沉淀,再通过DEAE-Dextran-Gel A-25柱层析分离和Sephadex G-200柱层析纯化,得到三种白色粉末状的多糖,命名为TF-A、TF-B及TF-C。用聚丙烯酰胺凝胶电泳,葡聚糖凝胶柱层析及气相色谱分析证明三者均为均一体。TF-A、TF-B及TF-C用酸完全水解,经纸层析和气相色谱分析表明:TF-A由L-岩藻糖、L-阿拉伯糖、D-木糖、D-甘露糖、D-半乳糖和D-葡萄糖组成,摩尔比为葡萄糖:甘露糖:木糖:岩藻糖:阿拉伯糖:半乳糖=1.06:1.0:0.33:0.29:0.037:0.75,TF-B及TF-C都由L-岩藻糖、L-阿拉伯糖、D-木糖、D-甘露糖、D-葡萄糖和葡萄糖醛酸组成,摩尔比分别为:葡萄糖:甘露糖:木糖:岩藻糖:阿拉伯糖:葡萄糖醛酸=0.16:1.0:0.28:0.73:0.036:0.19和0.086:1.0:0.37:0.75:0.058:0.37。     

蹄叶槖吾叶多糖提取工艺优化及单糖组成研究

采用水提醇沉法提取蹄叶槖吾叶粗多糖,通过单因素试验和Box-Benhnken法优化该多糖的提取工艺。结果表明:蹄叶槖吾叶粗多糖的最佳提取工艺为:料液比1∶20(g/m L)、提取温度90℃、提取时间2 h、提取3次,提取率为8.76%;通过红外光谱扫描检测其特征基团,并应用高效液相色谱技术对已脱除蛋白的多糖结构进行初步鉴定,得到其单糖组成为半乳糖醛酸∶鼠李糖∶半乳糖∶阿拉伯糖∶甘露糖∶葡萄糖∶葡萄糖酸∶岩藻糖=43.5∶6.0∶23.6∶11.5∶6.3∶5.4∶1.6∶0.9。     

宁夏不同地区灵武长枣果实多糖的单糖成分分析

以宁夏4个不同地区(灵武、中宁、青铜峡、银川)成熟期的灵武长枣果实为研究对象,经水提醇沉法提取,采用DEAE-cellulose52和HW-55S分离纯化,并利用GC-MS法进行多糖的单糖组成分析。结果表明:多糖提取率最高的是灵武地区,达到1.795%;分离纯化后,4个地区的长枣多糖各得到1个中性(Ju-0)和3个酸性组分(Ju-1、Ju-2、Ju-3),其中Ju-2含量最高;GC-MS分析可知灵武长枣多糖含有阿拉伯糖、鼠李糖、核糖、岩藻糖、木糖、甘露糖、半乳糖、葡萄糖、葡萄糖醛酸、半乳糖醛酸10种单糖,不含果糖,以阿拉伯糖、核糖、半乳糖和2种糖醛酸为主,木糖含量最低。各地区多糖的单糖组成、含量各不相同,从各组分来看,四个地区多糖的Ju-0和Ju-1组分组成均以阿拉伯糖、核糖、半乳糖为主,四个地区多糖的组成差异主要在于Ju-2和Ju-3组分。从各地区单糖总量来看,灵武地区是阿拉伯糖含量最高,中宁、青铜峡、银川地区以葡萄糖醛酸含量为最高。     

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。     

大鲵皮肤粘液多糖的提取及单糖组成分析北大核心CSCD

本文对大鲵皮肤粘液多糖的提取和单糖组分进行了研究。采用水提取、碱提取、酸提取及酶提取法从大鲵皮肤粘液提取粗多糖,通过1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生高效液相色谱法分析大鲵粘液多糖的单糖组分。结果表明:碱提取、酸提取、水提取、碱性蛋白酶及酸性蛋白酶提取物的总糖含量分别为4.23%、1.54%、1.81%、3.71%、2.19%;碱提取物的单糖组分为甘露糖(Man)、葡萄糖醛酸(Glc UA)、半乳糖醛酸(Gal UA)、氨基葡萄糖(Glc N)、葡萄糖(Glc)、半乳糖(Gal);酸提取物的单糖组分为Man、Glc UA;水提取物的单糖组分为Man、Glc UA、Gal;碱性蛋白酶提取物的单糖组分为Man、Glc UA、Glc N;酸性蛋白酶提取物的单糖组分为Man、Glc UA、Glc,表明不同提取工艺得到的粘液多糖的单糖组成存在差异。本文为大鲵皮肤粘液多糖的开发利用提供了依据。     

大鲵皮肤粘液多糖的提取及单糖组成分析

本文对大鲵皮肤粘液多糖的提取和单糖组分进行了研究。采用水提取、碱提取、酸提取及酶提取法从大鲵皮肤粘液提取粗多糖,通过1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生高效液相色谱法分析大鲵粘液多糖的单糖组分。结果表明:碱提取、酸提取、水提取、碱性蛋白酶及酸性蛋白酶提取物的总糖含量分别为4.23%、1.54%、1.81%、3.71%、2.19%;碱提取物的单糖组分为甘露糖(Man)、葡萄糖醛酸(Glc UA)、半乳糖醛酸(Gal UA)、氨基葡萄糖(Glc N)、葡萄糖(Glc)、半乳糖(Gal);酸提取物的单糖组分为Man、Glc UA;水提取物的单糖组分为Man、Glc UA、Gal;碱性蛋白酶提取物的单糖组分为Man、Glc UA、Glc N;酸性蛋白酶提取物的单糖组分为Man、Glc UA、Glc,表明不同提取工艺得到的粘液多糖的单糖组成存在差异。本文为大鲵皮肤粘液多糖的开发利用提供了依据。     

茶叶中酸性杂多糖的部分化学性质及降血糖活性的研究

粗老绿茶用醇提、复合纤维素酶提取、醇沉、再经弱碱性的大孔阴离子交换树脂D315柱层析,经NaCl洗脱,超滤除盐得酸性多糖ATPS。传统多糖的分离纯化一般采用价格昂贵的DEAE纤维素柱层析,很难进行工业化生产,为考察价格便宜的大孔阴离子交换树脂D315的分离纯化性能,对其分离得到的酸性茶多糖ATPS的部分化学性质进行了研究。将ATPS上DEAE52纤维素柱进一步纯化,收集DEAE52 NaCl梯度洗脱的主峰,透析,冻干,得ATPS-Ⅰ。高效凝胶渗透色谱分析发现ATPS与ATPS-Ⅰ分子量分布基本一致,气相色谱和离子色谱分析表明两者的单糖组成均以鼠李糖、阿拉伯糖、半乳糖和半乳糖醛酸为主,而且摩尔比近似。紫外光谱扫描表明ATPS不含蛋白质,红外光谱分析表明其糖苷键键型有α-型和β-型两种,单糖主要以吡喃糖苷形式存在。实验表明大孔阴离子交换树脂D315可以起到分离纯化茶多糖的作用。动物实验表明：ATPS能抑制四氧嘧啶糖尿病小鼠血糖的升高。     

地木耳多糖的提取与纯化研究

对地木耳采用水提醇沉法获得的地木耳多糖粗提取物,采用Sevage法脱蛋白质、醇沉,干燥得粗多糖,进一步用DEAE-52纤维素柱层析分离纯化,用纸色谱和琼脂糖凝胶电泳对洗脱组分进行纯度鉴定。结果表明：Sevage法脱蛋白7次可脱除94%的蛋白质,多糖得率为13.75%。DEAE-52纤维素柱层析后得到10种组分,浓缩干燥后得到白色粉末状多糖组分,每个组分经过纯度鉴定后均为单一的多糖。选择水和NaC l溶液为洗脱剂的温和条件分离纯化多糖效果较好。     

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。     

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。     

Structure of a 2-aminoethyl phosphate-containing O-specific polysaccharide of Proteus penneri 8 from a new serogroup O67.

An acidic O-specific polysaccharide was obtained by mild acid degradation of the Proteus penneri 8 lipopolysaccharide and found to contain D-glucose, D-galacturonic acid, 2-acetamido-2-deoxy-D-glucose, 2-acetamido-2-deoxy-D-galactose, 2-acetamido-2,6-dideoxy-L-galactose (L-FucNAc) and 2-aminoethyl phosphate (PEtn) in the ratios 2 : 1 : 1 : 1 : 1 : 1. 1H and 13C NMR spectroscopy was applied to the intact and dephosphorylated polysaccharides, and the following structure of the hexasaccharide repeating unit was established: The O-specific polysaccharide has a unique structure, and, accordingly, we propose for P. penneri 8 a new Proteus O67 serogroup, in which this strain is at present the single representative. The nature of epitopes on LPS of P. penneri 34, P. mirabilis O16, P. mirabilis O23 and P. vulgaris O22, which cross-react with O-antiserum against P. penneri 8, is discussed.     

Induction and Inducers of the Pectolytic System in Aureobasidium pullulans

The strain of Aureobasidium pullulans NRRL Y-2311 (CCY 27-1-98), known as a hyperproducer of endo-1,4-β-xylanase, exhibited good growth on pectin or pectate. Growth on these carbon sources is associated with an inducible production of significant amounts of pectolytic enzymes, of which exopolygalacturonase (EC 3.2.1.67) and endopolygalacturonase (EC 3.2.1.15) were identified. The two enzymes are not produced on D-glucose or under carbon starvation conditions. The enzymes can be induced in glucose-grown cells by D-galacturonic acid and its oligomers. Thus, D-galacturonic acid, the monomer derived from the polysaccharide, appears to be the natural inducer or a precursor of an inducer of pectolytic enzymes in the studied yeast. Received: 4 November 1995 / Accepted: 11 December 1995     

A yellow polysaccharide-producing bacterium with unusual characteristics

Summary A polysaccharide-producing Gram-negative bacterium was isolated from a sample of hay. It grew best on nitrate-containing media with sucrose as carbon source; the colony form was highly unusual. No polysaccharide was formed on glucose. A spontaneous mutant producing the same polysaccharide on both sucrose- and glucose-containing media was isolated. The polysaccharide has been characterized as an acid heteropolymer containing D-galacturonic acid, D-glucose, D-galactose and D-mannose in the approximate molar ratio 1:1:3:1, together with about 2 mol of acetate. The polysaccharide in aqueous solution was highly viscous with pseudoplastic characteristics.     

Isolation using triflic acid solvolysis and identification of N(epsilon)-[(R)-1-carboxyethyl]-N(alpha)-(D-galacturonoyl)-L-lysine as a component of the O-specific polysaccharide of Proteus mirabilis O13

An amino acid was released from the O-specific polysaccharide of Proteus mirabilis O13 by acid hydrolysis and identified as N(epsilon)-[(R)-1-carboxyethyl]-L-lysine by comparison with the authentic sample. An amide of this amino acid with D-galacturonic acid was isolated from the polysaccharide by solvolysis with anhydrous trifluoromethanesulfonic (triflic) acid and characterised by 1H and 13C NMR spectroscopy. These and published data enabled determination of the full structure of the repeating unit of the polysaccharide.     

Lipopolysaccharides from Pseudomonas maltophilia: composition of the lipopolysaccharide and structure of the side-chain polysaccharide from strain N.C.I.B. 9204

Lipopolysaccharide was extracted from defatted cell-walls of Pseudomonas maltophilia N.C.I.B. 9204. The major fatty acid components were 9-methyldecanoic acid, 2-hydroxy-9-methyldecanoic acid, 3-hydroxy-9-methyldecanoic acid, 3-hydroxy-dodecanoic acid, and 3-hydroxy-11-methyldodecanoic acid. Monosaccharide components of the phosphorylated core-oligosaccharide were D-glucose, D-mannose, D-galacturonic acid, 2-amino-2-deoxyglucose, and a 3-deoxyoctulosonic acid. The putative O-specific polysaccharide was composed mainly of 2-amino-2-deoxy-D-glucose, D-arabinose, and 6-deoxy-L-talose, but also contained an O-acetyl group and small proportions of rhamnose and 6-deoxy-3-O-methyltalose. Degradative and n.m.r. (1H and 13C) studies showed that the polymer had a branched trisaccharide repeating-unit with the following structure; the O-acetyl group was tentatively assigned to C-2 of the 6-deoxytalopyranosyl residue. (Formula: see text).     

Structure of the O-specific polysaccharide of the bacterium Proteus vulgaris O23

An acidic O-specific polysaccharide was obtained by mild acid degradation of the lipopolysaccharide of the bacterium Proteus vulgaris O23 (strain PrK 44/57) and found to contain 2-acetamido-2-deoxy-D-galactose, 2-acetamido-2-deoxy-D-glucose, and D-galacturonic acid. Based on 1H- and 13C-NMR spectroscopic studies, including two-dimensional correlation spectroscopy (COSY), total correlation spectroscopy (TOCSY), nuclear Overhauser effect spectroscopy (NOESY), and 1H,13C heteronuclear multiple-quantum coherence (HMQC) experiments, the following structure of the branched tetrasaccharide repeating unit of the polysaccharide was established: [figure], where the degree of O-acetylation of the terminal GalA residue at position 4 is about 80%. A structural similarity of the O-specific polysaccharides of P. vulgaris O23 and P. mirabilis O23 is discussed.     

Characteristic of Polysaccharide Complexes from <Emphasis Type="Italic">Centaurea scabiosa</Emphasis> L. and <Emphasis Type="Italic">Centaurea pseudomaculosa</Emphasis> Dobrocz.

We characterized polysaccharide complexes from Centaurea scabiosa L. and Centaurea pseudomaculosа Dobrocz. We proposed the technique of sequential selection of water-soluble polysaccharides and pectin substances from the aerial parts of studied objects. We have discovered that the content of water-soluble polysaccharides in the aerial parts of C. scabiosa was 2.8 times higher (2.7 ± 0.3%, n = 3) than in C. pseudomaculosа (0.97 ± 0.50%, n = 3). The content of pectin substances in the aerial parts of C. scabiosa was 2 times higher (7.6 ± 0.4%, n = 3) than in C. pseudomaculosа (3.9 ± 0.3%, n = 3). The residues of D-galacturonic acid, L-rhamnose, D-xylose, D-mannose, D-glucose, and D-galactose are the monomeric units of polysaccharide complexes from C. scabiosa and C. pseudomaculosa. Using ion-exchange chromatography, three polysaccharide fractions (molecular weights 667, 722, and 1027 kDa), whose monomer units are D-galacturonic acid, L-rhamnose, D-galactose, D-xylose, and D-glucose were isolated from the water-soluble polysaccharides of C. scabiosa.     

Chemical characterization of the lipopolysaccharides from enteropathogenic Escherichia coli O142 and O158.

Lipopolysaccharides (LPS) from two enteropathogenic strains of E. coli O142 and O158 were isolated by hot phenol-water extraction procedure. Polyacrylamide gel electrophoretic pattern of the LPS showed the typical ladder like pattern of smooth type of LPS. The LPS of E. coli O158 was found to contain L-rhamnose, D-glucose and N-acetyl-D-galactosamine as major constituents together with D-galactose, N-acetyl-D-glucosamine, L-glycero-D-manno-heptose and 2-keto-3-deoxy-D-manno-octulosonic acid (KDO) whereas LPS from E. coli O142 contained L-rhamnose, N-acetyl-D-glucosamine and N-acetyl-D-galactosamine as major constituents together with D-glucose, D-galactose, N-acetyl-D-glucosamine, L-glycero-D-mannoheptose and 2-keto-3-deoxy-D-manno-octulosonic acid (KDO). LPS was degraded by mild acid hydrolysis to yield a degraded polysaccharide fraction and an insoluble lipid-A fraction. The main fatty acids of the lipid-A fraction of the LPS were C12:O, C14:O, and 3-OH C14:O for O158 strain whereas E. coli O142 lipid-A consisted of C12:O, C14:O, 3-OH C14:O, and C16:O. The degraded polysaccharide fraction on gel permeation chromatography gave a high moleculer weight O-chain fraction and a core oligosaccharide and a fraction containing degraded sugars. The chemical composition of LPS and its fragmented products are reported in this communication.     

The structure of the specific capsular polysaccharide of Rhodococcus equi serotype 4

The specific capsular polysaccharide produced by Rhodococcus equi serotype 4 was found to be a high-molecular-weight acidic polymer composed of D-glucose, D-mannose, pyruvic acid and a previously unidentified 5-amino-3,5-dideoxynonulosonic (rhodaminic) acid in the proportions 2:1:1:1. Structural analysis, employing a combination of microanalytical methods, nuclear magnetic resonance spectroscopy, and mass spectrometric techniques, established that the polysaccharide consisted of linear repeating tetrasaccharide units having the sequence of residues shown below. In the native polysaccharide, the rhodaminic acid residues were present as their acetamido derivatives (RhoANAc) and carried 1-carboxyethylidene groups that bridged the O-7 and O-9 positions. Treatment of the capsular polysaccharide with dilute acetic acid and/or anhydrous hydrogen fluoride under hydrolytic/solvolytic conditions, resulted in the formation of four different oligosaccharide species. The 1H and 13C NMR resonances of these oligosaccharide fragments and of the native serotype 4 capsular polysaccharides were fully assigned by homo- and heteronuclear chemical shift correlation methods.     

Structure of an acidic O-specific polysaccharide from marine bacterium Shewanella fidelis KMM 3582T containing Nepsilon-[(S)-1-carboxyethyl]-Nalpha-(D-galacturonoyl)-L-lysine

The O-specific polysaccharide was isolated by mild acid degradation of the lipopolysaccharide of the marine bacterium Shewanella fidelis type strain KMM 3582T and studied by sugar analysis along with 1H and 13C NMR spectroscopy including one-dimensional NOE in difference mode and two-dimensional experiments. The polysaccharide was found to consist of linear tetrasaccharide repeating units containing Nepsilon-[(S)-1-carboxyethyl]-Nalpha-(D-galacturonoyl)-L-lysine and having the following structure: [See text.] The amide of D-galacturonic acid with Nepsilon-[(S)-1-carboxyethyl]-L-lysine ('alaninolysine', 2S,8S-AlaLys) was found for the first time in nature as a component of the O-specific polysaccharide of Providencia rustigianii O14 (Carbohydr. Res. 2003, 338, 1009-1016).