多糖高级结构解析方法的研究进展

研究发现,多糖的生物活性不仅与其一级结构（如分支度、单糖组成及糖苷键连接方式等）有关,而且与其空间结构也密不可分。 因此,对多糖高级结构的研究将有助于阐释其构效关系,为寻找具有生物活性的多糖和开发多糖药物提供理论依据。传统的多糖结构分 析方法具有一定的局限性,已不能满足多糖研究的需求,而现代物理技术的发展则为多糖高级结构解析提供了新的手段。综述近年来多 糖高级结构解析方法的研究进展。     

聚酰胺树脂分离纯化芦荟多糖的研究

文章采用聚酰胺吸附树脂,采用水、0．2M／LNaCl溶液、5％NaOH水溶液为洗脱剂梯度洗脱分离纯化,得到了三个芦荟多糖组分,通过IR、UV、GC等手段对其进行了分析。以甘露糖为标准,采用硫酸．苯酚法,测得水洗和NaOH水溶液洗脱所得多糖含量分别为90．10％和93．28％;以木糖、甘露糖、葡萄糖、半乳糖为单糖标准的GC分析结果显示：水洗所得多糖主要以甘露糖为主,同时含有少量的葡萄糖。     

Comparative characterization of polysaccharide fractions in grapes]

Different methods of isolating polysaccharides from grapes were evaluated with respect to the three indictrial grape varieties Traminer pink, Rkatsiteli, Cabernet-Sovinjon. Hot water extration was shown to be the best method in the isolation of water-soluble polysaccharides in primary wine-making. The qualitative and quantitative composition of the isolated polysaccharides was found to depend on the method of isolating polysaccharide fractions. The polysaccharides of grapes were heterogeneous in their molecular weight. The results of periodate oxidation of high molecular weight carbohydrates suggest a branched structure of polymers of water-soluble polysaccharides of grapes.     

Use of Methylotrophic Bacteria Methylobacillus flagellatumKT for Isolation of Deuterated Exogenous Carbohydrates

Cultivation conditions optimal for biosynthesis of exogenous polysaccharides by methylotrophic bacteria Methylobacillus flagellatum KT were evaluated. The mutant strain most active in accumulating exogenous polysaccharides was selected. Gradual adaptation of this strain to the deuterated medium containing 1 vol % CD₃OD in deuterium oxide intensified biosynthesis of exogenous polysaccharides and inhibited bacterial growth (compared to the standard medium). The monomeric composition of exogenous polysaccharides obtained during cultivation on standard and deuterated media was estimated by HPLC and NMR spectroscopy. Nondeuterated exogenous polysaccharides contained only fructose, whereas deuterated exogenous polysaccharides contained 98% fructose and 2% ribose. Paramagnetic resonance spectroscopy showed that the degree of deuterium incorporation into molecules of biosynthetic carbohydrates was 89%.     

Use of methylotrophic bacteria Methylobacillus flagellatum KT for isolation of deuterated exogenous carbohydrates

The cultivation conditions optimal for biosynthesis of exogenous polysaccharides by methylotrophic bacteria Methylobacillus flagellatum were evaluated. The mutant strain most active in accumulating exogenous polysaccharides was selected. Gradual adaptation of this strain to the deuterated medium containing 1 vol % CD3OD in deuterium oxide intensified biosynthesis of exogenous polysaccharides and inhibited bacterial growth (compared to the standard medium). The monomeric composition of exogenous polysaccharides obtained during cultivation on standard and deuterated media was estimated by high-performance liquid chromatography and nuclear magnetic resonance spectroscopy. Nondeuterated exogenous polysaccharides contained only fructose, while deuterated exogenous polysaccharides included 98% fructose and 2% ribose. Paramagnetic resonance spectroscopy showed that the degree of deuterium incorporation into molecules of biosynthetic carbohydrates was 89%.     

平菇菌粗多糖的抗氧化活性研究

采用深层发酵技术生产平菇粗多糖,时其清除DPPH自由基、羟自由基的能力、铁离子螯合能力以及还原力进行了比较分析。结果表明：菌丝体粗多糖和发酵液粗多糖均具有较强的抗氧化能力,但2种多糖的抗氧化能力存在差异;茵丝体粗多糖清除DPPH自由基的能力较强,其EC。。值为2．20mg／mL;发酵液粗多糖清除羟自由基的能力、铁离子螯合能力以及还原力较强,其EC50值分别为0．72mg／mL、3．32mg／mL和7．91mg／mL。在一定的浓度范围内,多糖的浓度增加,其抗氧化能力也随之增强,呈量效依赖关系。     

Extraction of polysaccharides and the antioxidant activity from the seeds of Plantago asiatica L

The extraction conditions of polysaccharides from Plantago asiatica L. seeds were investigated. Four parameters affecting the polysaccharides extraction, extraction times, water to sample, extraction temperature and single extraction time, were determined by orthogonal experiments. Under the optimized conditions, the polysaccharides yield of P. asiatica L. seeds was 2.467%. The antioxidant activities of the polysaccharides were investigated. The reducing power of the polysaccharides was dose dependent, and the reducing capacity of the polysaccharides was inferior to butylated hydroxytoluene, which is known to be a strong reducing agent. The scavenging rates of the polysaccharides on superoxide and 1,1-diphenyl-2-picrylhydrazyl radicals were79.7% and 81.4%, at polysaccharides concentration of 0.75 mg/mL, respectively, a scavenging rates approximately similar to that of 0.75 mg/mL ascorbic acid (83.5% and 85.1%, respectively). Furthermore, it exhibited a moderate concentration-dependent ABTS radical scavenging activity, ferrous ion chelating potency and H₂O₂ scavenging activity. The data obtained in the in vitro models clearly establish the antioxidant potency of the polysaccharides extracted from Semen Plantaginis.     

海藻多糖生物活性及分子修饰

简要介绍了近年来有关海藻多糖抗病毒、抗肿瘤、免疫促进、免疫抑制等生理活性的最新研究 ,重点介绍了不同种类海藻多糖的不同生理活性机理的研究进展。对多糖与生理活性之间的构效关系进行了阐述 ,在构效关系基础上进行多糖分子结构修饰是提高海藻多糖生理活性、降低毒副作用的有效手段。进一步介绍了目前多糖分子修饰常用方法 ,并对修饰后分子的生理活性改变进行了阐述。     

杏鲍菇菌丝体水溶性多糖提取及培养条件优化

以马铃薯葡萄糖综合培养基(PDP)为基础培养基,采用正交试验法优化杏鲍菇菌丝体多糖发酵条件,对接种量、摇床转速和培养时间等因素对多糖含量的影响进行了研究。采用水提醇沉法提取多糖,苯酚—浓硫酸法进行多糖含量测定。结果表明,最佳培养条件:接种量为每瓶1块直径为1cm的菌块,转速为140r/min,培养时间为8d。此时杏鲍菇菌丝体多糖含量最高,为75.1mg/g。     

Changes in Morphology of J774.1 Macrophage-like Cells and Release of Prostaglandin E2 Induced by Bacterial Acidic Polysaccharides

In an attempt to finding polysaccharides with immunostimulating activity, we isolated 14 bacteria that produced polysaccharides from soil. The polysaccharides produced by these bacteria and by strains kept in this laboratory and commercial polysaccharides were tested for ability to activate J774.1 macrophage-like mouse cells. Acidic polysaccharides, especially those from the newly isolated bacteria, induced spreading of the cells and release of prostaglandin E₂ from the cells, which are manifestations of a state of activation. The prostaglandin E₂-releasing activity of the polysaccharides was 1/10 to 1/20 on a weight basis of that of Escherichia coli lipopolysaccharide, a potent immunostimulant.     

密枝瑚菌多糖提取工艺参数优化

本文对密枝瑚菌多糖的测定方法和提取工艺参数进行了研究。结果表明,采用硫酸苯酚法测定密枝瑚菌多糖方法简便、准确、灵敏度高、重现性好。在此基础上采取单因素实验和正交分析,得到密枝瑚菌多糖最佳提取工艺参数为料液比1∶20,提取时间3 h,提取温度90℃。在此条件下,多糖提取率为2.23%。     

Effect of Sulfated Modification on the Molecular Characteristics and Biological Activities of Polysaccharides from Hypsizigus marmoreus

The effect of sulfated modification on polysaccharides from Hypsizigus marmoreus was examined by determining their molecular structures and bioactivities. The sulfation, which was implemented by using an orthogonal array design, produced polysaccharides with varying degrees of substitution (DS) ranging from 0.11 to 1.06. The sulfated polysaccharides exhibited a lower average molecular weight (M _w) and considerably higher radius of gyration (R _g) than those of native polysaccharide, suggesting that the conformation of the sulfated polysaccharides had been changed towards a more extended type. The inhibitory activity toward cancer cell growth was enhanced by treating with the sulfated polysaccharides by up to 34%, as compared to the native polysaccharide. In addition, treating with the sulfated polysaccharides increased the nitric oxide (NO) and cytokine (IL-1β and TNF-α) release to levels comparable to those detected in the positive control, lipopolysaccharide (LPS), suggesting that the sulfated polysaccharides might have strong immunomodulatory activity.     

枸杞色素及多糖的不同提取次序对其得率及抗氧化活性的影响

为探究先后提取枸杞多糖及枸杞色素时对各自得率的影响。本研究通过分别考察这两种成分在提取过程中的提取溶剂、提取温度、料液比、提取次数及提取时间等因素对枸杞色素和枸杞多糖的得率影响,确定枸杞色素和枸杞多糖在提取次序不同时,两者的最佳提取工艺以及对DPPH·和·OH自由基清除率。结果表明,首先提取枸杞多糖后,枸杞色素的最佳提取工艺为采用正己烷,80℃时,料液比1∶10,提取2次,每次1 h,枸杞色素得率为2.48%,此时枸杞多糖得率为7.45%;而首先提取枸杞色素后,采用了超声辅助提取的方式提取枸杞多糖,发现超声效率为25%,料液比1∶10,提取20 min,枸杞多糖得率为5.23%,此时枸杞色素得率为3.93%。因此,首先提取枸杞多糖,使其平均得率为7.45%,而后提取枸杞色素,其平均得率为2.48%;总体上,枸杞色素1和枸杞多糖1对DPPH·自由基清除率都较高,枸杞多糖1对·OH自由基清除率较高,其抗氧化活性都接近Vc。     

奶油绚孔菌菌丝多糖的提取及其抗氧化活性

为了获得奶油绚孔菌菌丝多糖的最佳制备工艺并进一步评估其体外抗氧化的作用,本研究以奶油绚孔菌菌丝作为原料,采用中心组合试验法结合响应面分析法对奶油绚孔菌菌丝多糖的提取条件进行优化。结果表明,最佳的提取工艺为：料液比1:32、提取时间1.85 h和提取温度85 ℃,奶油绚孔菌菌丝多糖在此提取工艺条件下的得率为5.68%。奶油绚孔菌菌丝多糖对DPPH、ABTS和羟基自由基有清除能力,其EC₅₀分别为3.74、3.80和15.10 mg/mL。奶油绚孔菌菌丝多糖呈现出一定的还原能力。由此可知,奶油绚孔菌菌丝多糖具有一定的体外抗氧化作用,该研究能够为奶油绚孔菌菌丝多糖的进一步综合开发提供理论支持。     

乌梅丸水提液中多糖理化性质分析及预防结肠炎的作用

目的:考察乌梅丸水提液中多糖含量,建立HPLC法分析乌梅丸多糖分子量及单糖组成方法,观察其对右旋葡聚糖硫酸钠(DSS)所致小鼠溃疡性结肠炎(UC)的预防作用,为乌梅丸临床用药及药理活性成分的研究提供依据。方法:采用水提醇沉法提取多糖;硫酸-苯酚法测定乌梅丸多糖含量;凝胶色谱法-龙智达分子量工作站测定多糖分子量;PMP柱前衍生化法HPLC分析单糖组成;DSS(3%)法建立UC小鼠模型。ICR小鼠50只随机分为5组:正常组,模型组,乌梅丸多糖2.5%,5%,10%剂量组,每组10只。造模前7天,在乌梅丸多糖治疗各组的小鼠饮水中添加乌梅丸多糖,一直给药维持至实验结束(第14天)。结果:葡萄糖在0~0.08 mg·m L-1浓度范围内呈良好的线性关系(r=0.998);右旋葡聚糖酐标准品分子量在2500~2000000 Da范围内呈良好线性关系(r=0.998);乌梅丸水提液中多糖含量为40.3%,纯化的乌梅丸多糖糖含量为91.6%;分子量范围在171343~525009 Da之间,分布系数D=1.18;其多糖的单糖组成主要为甘露糖、葡萄糖醛酸、葡萄糖、半乳糖和木糖组成,其相对含量比为1.8:1.0:19.3:32.8:4.2;乌梅丸多糖能降低结肠组织损伤程度。结论:多糖可能是乌梅丸汤剂中有效的活性成分,对小鼠UC有一定的防治作用,其理化性质分析方法简便快速,结果准确,重复性好,可作为乌梅丸多糖的质量控制方法。     

红托竹荪菌托多糖的提取及抗肿瘤活性的初步研究

红托竹荪菌托经热水提取、酒精沉淀、脱蛋白后的粗多糖,其得率远大于其菌丝体和子实体的其他部位及香菇子实体.菌托粗多糖进一步用DEAE纤维素柱和Sephadex G-75分离纯化,得到两个组分DRVP1与DRVP2,对分离得到的主要多糖通过高效液相色谱(HPLC)、红外光谱(IR)等进行结构分析.DRVP1的相对分子质量(Mr)为1.47×104,红外光谱数据显示为β-型甘露糖苷.体外试验表明,红托竹荪菌托多糖的组分DRVP1对小鼠S180肉瘤有一定的抑制作用.     

Replication-competent gamma-retrovirus Mo-MuLV expressing green fluorescent protein as efficient tool for screening of inhibitors of retroviruses that use heparan sulfate as primary cell receptor

The effect of sulfated polysaccharides on the efficiency of the infection of mouse transplantable fibroblast SC-1 and NIH-3T3 cell lines by replication-competent recombinant Moloney murine leukemia virus (Mo-MuLV), which carries the eGFP gene, was investigated. It was found that sulfated polysaccharides have no cytostatic and cytotoxic effects on SC-1 and NIH-3T3 cells in the concentration range of 0.01–100 μg/mL and have virucidal activity against Mo-MuLV. Polysaccharides in the indicated concentrations inhibit Mo-MuLV infection that prevents the further development of viral infection. It was shown that sulfated polysaccharides are also effective inhibitors of other retroviruses, including lentiviruses, which use sulfated polysaccharides as primary cell receptors.     

Immunochemical analysis of the types Ia and Ib group B streptococcal polysaccharides

The types Ia and Ib group B streptococcal type-specific polysaccharides have remarkable immunologic differences despite a great deal of structural similarity. Although these two complex polysaccharides differ only by a single glycosidic linkage, they are antigenically distinct. Furthermore, terminal sialic acid residues appear to be critical to the immunodeterminant on the type Ia polysaccharide, whereas the antigenicity of the type Ib polysaccharide does not show this dependence on sialic acid. In the current investigation we defined better the immunodeterminant of these polysaccharides. With homologous rabbit antiserum, the type Ia native and core polysaccharides demonstrated partial serologic identity, whereas the type Ib native and core polysaccharides demonstrated complete serologic identity. Surprisingly, the type I degalactosylated polysaccharide, degraded structure, was capable of reacting with a population of antibodies present in type Ia antiserum similar to the complete type Ia native polysaccharide, although demonstrating a reduced level of immunodeterminant expression. Unlike the reactions of the type Ia polysaccharides with homologous rabbit antiserum, the Ib native and core polysaccharides were able to react with identical populations of antibodies in type Ib-specific antiserum. A minor population of antibodies was demonstrated in the type Ib antiserum, which was reactive with the degalactosylated polysaccharide. That a population of antibodies reactive toward the degalactosylated polysaccharide is present in both type Ia and type Ib antisera suggests that the Iabc cross-reacting determinant is due to the presence of serum antibodies reactive with this trisaccharide repeating unit, which is shared by both the type Ia and the type Ib native and core polysaccharides.     

超滤膜技术在多糖提取方面的应用

超滤膜技术广泛应用于各类多糖的分离、浓缩、纯化等研究中,包括中药药源多糖如灵芝多糖、大黄多糖、六味地黄汤多糖、黄芪多糖、紫芝多糖、人参多糖,海洋活性多糖如鲨鱼软骨粘多糖、褶纹冠蚌多糖、紫菜多糖、褐藻糖胶、卡拉胶,发酵多糖如蜜环菌菌索多糖、PS-9415发酵液多糖、冬虫夏草多糖,食用植物多糖如茶多糖、香菇多糖、金针菇多糖、芦荟多糖、枸杞多糖等.采用超滤膜技术处理多糖具有收率高、不易破坏多糖的生物活性、能耗低等特点,适于工业化生产.     

忍冬桑黄和蛹虫草共发酵联产真菌多糖初步研究

忍冬桑黄和蛹虫草两种药用真菌均可产活性多糖,共发酵模式是产生新化合物或提高化合物含量或药效的潜在方式.本研究尝试用忍冬桑黄和蛹虫草共发酵联产真菌多糖,并对其共发酵所得的菌丝体多糖开展抗氧化活性试验.结果表明,当忍冬桑黄和蛹虫草预先分别发酵3d和1d后再同时接种共发酵,两菌可以较好地进行共生长,菌体总量和总多糖得率显著高...