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     

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。     

金刷巴多糖的研究

金刷巴用热水提取,乙醇沉淀,精制得ＬＥＣ多糖,经Ｓｅｐｈａｄｅｘ Ｇ－１５０柱层析均为一性多糖。气相气谱检测含有葡萄糖,甘露糖,克分子比为Ｇｌｃ：Ｍａｎ＝２７：１,平均分子量为１１Ｘ１０＾５,糖含量为８２．４５％。经红外光谱分析,高碘酸氧化,Ｓｍｉｔｈ降解,ＬＥＣ多糖主要是β（１→３）甙键聚合成而的多支链多糖。     

短裙竹荪多糖Dd—S3P的分离纯化及其性质研究

短裙竹荪子实体经２％Ｎａ２ＣＯ３溶液提取,用蛋白酶法和Ｓｅｖａｇ法相结合除去蛋白,乙醇分级沉淀,级分３经ＤＥＡＥ－ＳｅｐｈａｄｅｘＡ－２５柱层析纯化得到的短裙竹荪多糖Ｄｄ－Ｓ３Ｐ。经测定该多糖为均一组分,分子量约为３．８×１０＾５,红外光谱呈现出典型的多糖吸收峰,含有α－型糖苷连接键,紫外扫描无核酸和蛋白质的特征吸收峰。     

条斑紫菜中一种琼胶多糖的分离纯化及鉴定

条斑紫菜（Ｐｏｒｐｈｙｒａ ｙｅｚｏｅｎｓｉｓ）的热水提取物,经ＤＥ－２３和Ｓｅｐｈａｄｅｘ Ｇ－２００柱层析纯化,得到一种含微量硫酸基的琼胶精品（ＰＹ１）。此多糖经Ｓｅｐｈａｒｏｓｅ ６Ｂ柱层析鉴定为单一组分,分子量为２２０ｋＤ。紫外光谱显示它不含蛋白质、多肽及核酸。红外光谱揭示它含３,６－内醚－半乳糖的特征吸收以及微量的硫酸基。该多糖的水解产物经气相色谱分析,主要由半乳糖及其衍生物组成,有少量     

枸杞子糖蛋白的分离纯化、物化性质及糖肽键特征

从宁夏枸杞子提取得到的粗多糖,经ＤＥＡＥ－Ｃｅｌｌｕｌｏｓｅ和ＳｅｐｈａｄｅｘＧ－１００柱层析,得到均一的枸杞子糖蛋白ＬｂＧＰ。分子量由ＳＤＳ－ＰＡＧＥ测定为８８ｋｄ,糖含量为７０％,糖组成为Ａｒａ：Ｇａｌ：Ｇｌｃ＝２．５：１．０：１．０（摩尔比）,并含有其他１８种天然氨基酸。初步分析表明ＬｂＧＰ是Ｏ－连接的糖蛋白。     

深层培养云芝菌丝体蛋白多糖的提取及性质

采用水、３％草酸和０．２ｍｏｌ／Ｌ氢氧化钠为溶剂分别提取深层培养的云芝〔Ｐｏｌｙｓｔｉｃｔｕｓｖｅｒｓｉｃｏｌｏｒ（Ｌ．）Ｆｒ．〕菌丝体蛋白多糖,其得率分别为菌丝体干重的２０％、４９％和１９％。提取的粗多糖经ＳｅｐｈａｄｅｘＧ１００柱层析进一步分离、纯化,纯化的多糖经聚丙烯酰胺凝胶电泳分离后呈现一个斑点,在ＳｅｐｈａｄｅｘＧ１００柱层析中有一个对称峰,紫外及红外光谱分析和蛋白质含量测定表明该多糖为蛋白多糖,其蛋白质和多糖含量在水提多糖中分别为１３％和６２％、在酸提多糖中分别为５％和９１％。在碱提多糖中分别为３％和８１％。组成该多糖的主要单糖是葡萄糖和木糖。     

甲基单胞菌Methylomonas sp.GYJ3中甲烷单加氧酶还原酶组分的纯化和性质

Ｍｅｔｙｌｏｍｏｎａｓｓｐ．ＧＹＪ３菌的甲烷单加氧酶（ＭＭＯ）粗酶提取液经ＤＥＡＥ－ＳｅｐｈａｒｏｓｅＣＬ－６Ｂ阴离子交换层析、ＳｅｐｈａｄｅｘＧ－１００凝胶过滤层析和ＤＥＡＥ－ＴＳＫｇｅｌＨＰＬＣ分离纯化出ＭＭＯ还原酶组分．经ＨＰＬＣ分析,纯度大于９５％,纯化倍数为４．４,加入至ＭＭＯ羟基化酶和调节蛋白Ｂ的体系中表现比活为２２８ｎｍｏｌ环氧丙烷每分钟毫克蛋白．ＳＤＳ－ＰＡＧＥ电泳表明还原酶由一种亚基组成,分子量４２ｋＤ．ＩＣＰ－ＡＥＳ测定还原酶的Ｆｅ含量为１．８３ｍｏｌＦｅ每ｍｏｌ蛋白．ＵＶ－Ｖｉｓ光谱表明还原酶除２８０ｎｍ蛋白质特征峰外在４６０ｎｍ有最大吸收峰,且Ａ２８０ｎｍ／Ａ４６０ｎｍ为２．５０,与其它黄素一铁硫蛋白相似,推测还原酶可能含一个ＦＡＤ辅基和Ｆｅ２Ｓ２中心．在厌氧条件下,还原酶能够和ＮＡＤＨ作用,ＵＶ－Ｖｉｓ光谱分析表明还原酶４６０ｎｍ处特征吸收峰消失,说明在ＭＭＯ催化过程中还原酶接受ＮＡＤＨ的电子．ＤＥＡＥ－ＳｅｐｈａｒｏｓｅＣＬ－６Ｂ阴离子交换层析分离出调节蛋白Ｂ,部分纯化的调节蛋白Ｂ的分子量大约在２０ｋＤ,它能够提高ＭＭＯ比活性４０倍,ＭＭＯ还原酶和调节蛋白Ｂ单独存在时不具有ＭＭＯ     

枸杞子糖蛋白的分离纯化,物化性质及糖肽键特征

从宁夏枸杞子提取得到的粗多糖,经ＤＥＡＥ－Ｃｅｌｌｕｌｏｓｅ和ＳｅｐｈａｄｅｘＧ－１００柱层析,得到均一的枸杞子糖蛋白ＬｂＧＰ。分子量由ＳＤＳ－ＰＡＧＥ测定为８８ｋｄ,糖含量为７０％,糖组成为Ａｒａ：Ｇａｌ：Ｇｌｃ＝２．５：１．０：１．０,并含有其他１８种天然氨基酸。初步分析表明ＬｂＧＰ是Ｏ－连接的糖蛋白。     

地衣芽孢杆菌β—甘露聚糖酶的纯化及其性质的研究

地衣芽孢杆菌１Ｂａｃｉｕｕｓ Ｌｉｃｈｅｎｉｆｏｒｍｉｓ）ＢＬ－３０６产生的胞外β－甘露聚糖酶经硫酸铵分级盐析,ＤＥＡＥ－纤维素柱层析。Ｓｅｐｈａｄｅｘ－Ｇ１００柱凝胶过滤和ＤＥＡＥ－纤维素柱再层析分离纯化,得到ＳＤＳ－聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）均一样品。用ＳＤＳ－ＰＡＧＥ测得纯化后β－甘露聚糖酶分子量为２６０００道尔顿。用凝胶等电聚焦电泳（ＰＡＧＥＩＥＦ）测得等电点ＰＩ为５．０。该酶     

九里香多糖和蛋白多糖的分离、纯化和分析

九里香(Murraya Paniculata L.Jack)皮部经热水提取、乙醇沉淀、SephadexG—100柱层析分离、磷酸氢钙吸附、酸溶,再通过Sephadex G-200柱层析纯化,得到灰白色粉末状九里香蛋白多糖。总糖含量为52.1％(其中葡萄糖醛酸含量为10.9％),蛋白质含量为20.0％。九里香蛋白多糖去蛋白后,即得九里香多糖,总糖含量为88.2％(其中葡萄糖醛酸含量为20.0％),经纸层析和聚丙烯酰胺凝胶园盘电泳鉴定为单一斑点和单一区带。平均分子量约为1.7×10~(-5)。组成单糖的摩尔比为葡萄糖:露甘糖:木糖:阿拉伯糖:岩藻糖:葡萄糖醛酸=1.0:0.40:0.16:0.17:0.20:0.48.     

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     

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.     

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。     

Heterogenous Degradation of Myofibrillar Proteins

Pectic substances were extracted from the vegetables with oxalate buffer of pH 4.25 and, after saponification, fractionated into two components, weakly acidic pectic polysaccharide (WAP) and pectic acid, by DEAE-cellulose and Sephadex G-100 chromatographies. The galacturonic acid content (17.3~25.8%) of WAPs was much lower than that of pectic acids, though the neutral sugar compositions of both pectic substances were almost the same. The arabinose-galactose side chains were found to be very long or highly branched in WAPs compared with those in pectic acids.

All the WAPs were appreciably hydrolyzed by exo- and endopolygalacturonases. The limited-degradation products (the residual polysaccharides; i.e., the rhamnogalacturonan segments) obtained by endopolygalacturonase from both WAPs and pectic acids showed a similar behavior on Sephadex G-100 and Sepharose CL-4B gel filtrations; each of the rhamnogalacturonan segments was eluted in the void volume of the Sephadex G-100 column. From these results, we concluded that WAPs are probably an inherent pectic component of the cell walls of the vegetables.     

Polysaccharide covalently linked to the peptidoglycan of the cyanobacterium Synechocystis sp. strain PCC6714.

A polysaccharide was found to be covalently linked to the peptidoglycan of the unicellular cyanobacterium Synechocystis sp. strain PCC6714 via phosphodiester bonds. It could be cleaved from the peptidoglycan-polysaccharide (PG-PS) complex by hydrofluoric acid (HF) treatment in the cold (48% HF, 0 degrees C, 48 h) yielding a pure, HF-insoluble peptidoglycan fraction and an HF-soluble polysaccharide fraction. The PG-PS complex was isolated from the Triton X-100-insoluble cell wall fraction by hot sodium dodecyl sulfate treatment and digestion with proteases. Digestion of the complex with N-acetylmuramidase released the glycopeptide-linked polysaccharide, which was further purified by dialysis and gel filtration on Sephadex G-50 and G-200. The polysaccharide consisted of glucosamine, mannosamine, galactosamine, mannose, and glucose and had a molecular weight of 25,000 to 30,000. Muramic acid-6-phosphate was identified as the binding site of the covalently linked, nonphosphorylated polysaccharide as revealed by chemical analysis of linkage fragments of the PG-PS complex.     

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。     

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。     

Chemical properties of lipopolysaccharide (LPS) isolated from Vibrio anguillarum PT514

Vibrio anguillarum, one of the causative agents of fish vibriosis, is serologically and biochemically divided into three groups (A, B and C). The chemical composition and molecular architecture of lipopolysaccharide (LPS) isolated from V. anguillarum PT 514, which belongs to serogroup B, were investigated. The LPS contained glucose (Glc), fructose (Fru), L-glycero-D-mannoheptose (L-D Hep), glucosamine (GlcN) and 4-amino-4,6-dideoxyglucose as sugar constituents in molar ratios of 8.9:0.7:3.0:1.1:1.6. Sephadex G-50 gel-chromatography of a degraded polysaccharide fraction separated from the LPS by 5% acetic acid hydrolysis suggested that the O-specific polysaccharide region consists of, in average, as much as 29 moles of Glc per 3 moles of L-D Hep, while the core polysaccharide contains at least Glc, L-D Hep and GlcN in molar ratios of 3.2 : 3.0 : 0.2. Fru and 4-amino-4,6-dideoxyglucose components were released from LPS on weak-acid hydrolysis, indicating that PT 514 LPS is distinguishable from those of Vibrio anguillarum belonging to the other serogroups. 2-Keto-3-deoxyoctonate (KDO), a common sugar constituent of gram-negative bacterial LPS, was not detected by Weissbach's color reaction under the conventional hydrolysis condition, but O-phosphoryl KDO was found in the strong-acid hydrolysate (4 M HCl, 100 C, 45 min). This substance was identical, at least in high-voltage paper electrophoresis, to 5-O-phosphoryl KDO.(ABSTRACT TRUNCATED AT 250 WORDS)