洋甘菊多糖的分离纯化、性质结构及抗氧化活性分析

对热回流提取得到的洋甘菊多糖进行了分离纯化,并对得到的多糖的性质结构及抗氧化活性进行分析。经处理后的洋甘菊多糖先后利用DEAE-Sepharose和Sephadex G-100柱色谱分离纯化得到了2种均一多糖(MCP-1-1和MCP-2-1);经测定2种均一多糖总糖含量分别为82.23%、84.09%,糖醛酸含量分别为4.11%、5.38%,蛋白质含量分别为0.58%、1.05%,溶解度分别为98.36%、98.44%,分子质量分别为30902、7244 Da;单糖组成实验结果表明MCP-1-1是由鼠李糖(Rha)、葡萄糖醛酸(GlcA)、半乳糖(Gal)、葡萄糖(Glc)、木糖(Xyl)和阿拉伯糖(Ara)6种单糖组成,MCP-2-1是由GlcA、Gal、Glc、GalA、Xyl和Ara六种单糖组成,2种均一多糖均是不含三螺旋结构的酸性多糖,均含有吡喃环和β-糖苷键,MCP-1-1含有α-糖苷键;2种均一多糖具有良好的热稳定性。体外清除自由基试验结果显示MCP-1-1和MCP-2-1对DPPH自由基和·OH均有清除作用,而且清除能力与均一多糖的质量浓度正相关,MCP-2-1对2种自由基的清除能力明显高于MCP-1-1。研究结果表明,分离纯化得到的洋甘菊多糖可作为天然抗氧化剂,用于食品和化妆品中。     

Structural characterisation of a water-soluble polysaccharide with high branches from the leaves of Taxus chinensis var. mairei

A large number of polysaccharides present in the leaves of Taxus chinensis var. mairei were successively extracted with hot water after preliminary treatments. The ion-exchange and gel-permeation chromatography was used to isolate and purify the major polysaccharides to afford a complex water-soluble polysaccharide, named T1 with a molecular mass of 3.44 × 10⁶ Da determined by HPGPC. T1 consisted of 2,4-di-O-methyl-mannose, rhamnose, arabinose, xylose, galactose, mannose, galacturonic acid and glucuronic acid in a molar ratio of 2:5:24:9:3:46:1:10, and the acetyl content was estimated to be 2.1%. On the basis of methylation analysis, periodate and chromium trioxide oxidations, Smith degradation, graded acid hydrolysis, and NMR and IR spectroscopy, T1 possessed a 1,3,6-linked β-d-Manp main chain, substituted by mannose, arabinose, galactose, xylose, rhamnose, 2,4-di-O-methyl-mannose, galacturonic acid, and glucuronic acid residues. These contained non-reducing end-units of mannose, arabinose, xylose, 2,4-di-O-methyl-mannose, and glucuronic acid. This is the first report on isolation of an acid heteropolysaccharide from Taxus species.     

柱前衍生化HPLC法分析真菌多糖的单糖组成

通过柱前衍生化HPLC法分析了6种真菌多糖的单糖组成。将6种真菌的胞外粗多糖及其菌体用1mol/L的硫酸水解成单糖,用1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生化,利用高效液相色谱法检测各单糖。结果表明,灵芝、树舌类真菌胞外多糖的产率均较高,胞外多糖产率在0.0921～0.1528g/100mL之间。胞外多糖主要以甘露糖为主,其次含有葡萄糖和半乳糖,并且一般含有少量的阿拉伯糖。胞内多糖中葡萄糖含量最高,其次含有甘露糖和半乳糖,个别菌种还含有少量的阿拉伯糖和葡萄糖醛酸。而虫草胞外多糖的组成依次为半乳糖、葡萄糖、甘露糖和葡萄糖醛酸,胞内多糖同样是主要含有葡萄糖,其次含有甘露糖、半乳糖和葡萄糖醛酸。     

高效阴离子交换色谱法分析杜氏盐藻多糖的单糖组成

选用CarboPacPA20分析柱(3mmi.d.×150mm),以H2O-250mmol/LNaOH-1mol/LNaAc为流动相,建立了三元梯度洗脱分离、积分脉冲安培检测多糖中常见的8种中性单糖和2种糖醛酸的高效阴离子交换色谱分析方法;利用离子交换色谱柱(DEAESepharoseCL6Bfastflow,3.5cm×30cm)和凝胶过滤色谱柱(SepharoseCL6B,2.6cm×100cm)从提取出β-胡萝卜素的杜氏盐藻残渣中分离出PD1、PD2、PD3、PD4a和PD4b共5个多糖级分,选用适当的条件水解后,利用高效阴离子交换色谱法测定了各多糖级分的单糖组成。其中具有显著生物活性的多糖级分PD4a中主要含有半乳糖、阿拉伯糖、木糖、葡萄糖、盐藻糖及鼠李糖6种中性糖,还含有少量糖醛酸;PD4b中主要含有核糖、葡萄糖、半乳糖、阿拉伯糖及木糖5种中性糖,不含糖醛酸。该结果与PMP柱前衍生化反相高效液相色谱法测定的单糖摩尔比基本一致。10种单糖的离子色谱法的检出限为0.76～2.92nmol/L(以3倍信噪比计),峰面积相对标淮偏差RSD为0.72%～3.25%。     

玉米皮多糖的组成及结构

采用紫外光谱、纸层析及比色等分析方法对分离纯化制得的玉米皮多糖A-1(CSPA-1)、A-2(CSPA-2)、B(CSPB)的总糖含量、单糖组成及基本结构进行初步研究。结果表明：CSPA-1、CSPA-2 和CSPB 皆为白色粉末,其中CSPB 微溶于水,其他易溶于水,不溶于高浓度的有机溶剂,不含蛋白质和淀粉,均含有糖醛酸。CSPA-1总糖含量为99.3%,糖醛酸含量为16.6%,糖基组成为葡萄糖、木糖及阿拉伯糖,可能还含有微量鼠李糖;CSPA-2 总糖含量为93.4%,糖醛酸含量为21.7%,糖基组成为葡萄糖、木糖、阿拉伯糖及鼠李糖;CSPB 总糖含量为83.3%,糖醛酸含量为6.82%,糖基组成为葡萄糖、木糖及阿拉伯糖。高碘酸氧化分析表明：CSPA-1 中1 → 2 糖苷键或1 → 4 糖苷键残基比例为25%,1 → 6 糖苷键残基比例为3%;CSPA-2 中1 → 2 糖苷键或1 → 4 糖苷键残基比例为25.7%,1 → 6 糖苷键残基比例为3%;CSPB 中1 → 2 糖苷键或1 → 4 糖苷键残基比例为34.1%,1 → 6 糖苷键残基比例为1%。     

RELATIONSHIP BETWEEN LEVELS OF GIBBERELLIC ACID AND THE PRODUCTION AND ACTION OF CARBOHYDRASES OF BARLEY

Different hydrolytic enzymes require different levels of gibberellic acid to induce their maximal production and release into the endosperm of barley. Barley-endo-β-glucanase requires a higher level of gibberellic acid to induce maximal production than does α-amylase. Although gibberellic acid also increases the level of barley endo-β-1,3 glucanase, this enzyme, unlike the barley-endo-β-glucanase, develops to significant levels when gibberellic acid is absent. In gibberellic acid-treated aleurone layers β-glucanases degrade the cell wall mainly to glucose. Xylose and cellobiose appear when the aleurone wall has undergone extensive enzymic hydrolysis. Laminaribiose and arabinose are found whether or not gibberellic acid is present in the medium. In addition to the degradation of the endosperm cell walls, β-glucanases may also play an important role in the release of enzymes from the aleurone into the endosperm during malting.     

非肝素类天然抗凝血多糖的研究进展

血液凝固和血小板凝集是局部缺血性疾病发病的最重要的原因,诱发心血管和脑血管甚至全身各处的相关疾病。肝素类物质已被广泛应用于相关疾病的治疗,作用效果显著但副作用较大。本文介绍了非肝素类具有抗凝血功能的天然多糖类物质,其单糖组成主要是葡萄糖(醛酸)、半乳糖(醛酸)、阿拉伯糖、甘露糖、木糖、鼠李糖等,其中源于海洋的多糖均含有硫酸根,与多糖形成多糖硫酸酯类物质;而源于陆地的,一般不含硫酸根,含己糖醛酸或尿酸等酸性成分。不同来源的多糖其分子组成和结构差异较大,抗凝作用机理也不尽相同。本文着重阐述了海藻类、海洋无脊椎动物、植物及其果实、可食用菌等来源的抗凝血多糖结构特点、抗凝作用机理,希望能为高抗凝活性、低毒副作用的天然多糖类药物的进一步研究提供一定的参考。     

Cultivar and Maturity Effects on Muskmelon ( Cucumis melo) Colour,Texture and Cell Wall Polysaccharide Composition

Cell wall polysaccharides (CWP) of two types of melons were isolated and purified. Fractionations were performed using cyclohexanetrans-1,2-diamine tetraacetate (CDTA), Na₂CO₃, guanidinium thiocyanate (GTC) and KOH. Alditol acetate derivatives of neutral sugars from each CWP fraction were prepared and analysed by gas chromatography. Trifluoro-acetic acid insoluble fractions were analysed colorimetrically and uronic acid was determined. The CDTA and Na₂CO₃ fractions were found to be composed of typical pectic materialscontaining primarily galacturonic acid with the neutral sugars arabinose, galactose, rhamnose and a smaller amount of xylose. As maturity increased, CDTA fraction yields increased, though total neutral sugar CWP compositions decreased. GTC and KOH fractions were typical of hemicellulose, and contained principally xylose, glucose, galactose, mannose and fucose, with very small amounts of uronic acid, arabinose and rhamnose. The residues contained principally glucose and galactose, with smaller amounts of mannose, xylose, arabinose and fucose. With the exception of xylose and glucose, all neutral sugars decreased significantly during ripening in both the Cantaloupe and Honey Dew melons. Total uronic acid did not change as maturity increased, except for Cantaloupe, where total uronic acid decreased from the ripe to overripe stages. Relationships between firmness, drip loss and other composition measurements, as well as the total CWP sugar composition, were also determined. Only the CDTA fraction yields were negatively correlated with the changes in firmness of both melons and positively correlated with changes in drip loss as maturity increased.     

MORPHOLOGY AND CHEMICAL COMPOSITION OF BARLEY ENDOSPERM CELL WALLS

Cell walls have been isolated from barley endosperm and found to contain a microfibrillar phase which is embedded in an amorphous matrix. The microfibrillar phase probably consists of cellulose, together with tightly bound arabinoxylan and polysaccharides rich in mannose. The matrix material is arabinoxylan (approx. 25%) and β-glucan (approx. 75%). Pectic polysaccharides are absent from the isolated cell walls. After successive removal of the matrix polysaccharides with water and 1-M NaOH, only 6% of the wall remains. The intra-cellular surfaces of the wall fragments are extensively pitted, probably as a result of adpression of starch granules into the cell wall material during endosperm development. Although polysaccharides are the major components of the cell walls, some nitrogen (less than 1%) is present. Phenolic compounds may also be wall constituents, but hydroxyproline could not be detected.     

碱提西番莲叶多糖的分离、鉴定及生物活性

以热水提取后的西番莲叶残渣为原料,利用NaOH碱溶液提取多糖(APEL),通过离子交换层析进行分离,测定多糖的理化性质,同时采用红外光谱、高效液相色谱、凝胶渗透色谱法对多糖进行分析,并研究了西番莲叶中多糖组分的体外降血糖及抗凝血活性。结果表明:APEL经DEAE-52纤维素离子交换层析法分离得到3种组分:APEL-1、APEL-2、APEL-3,APEL及其分离组分均具有多糖的典型特征峰,主要由阿拉伯糖、葡萄糖、半乳糖、木糖、鼠李糖、葡萄糖醛酸组成,APEL、APEL-1、APEL-2和APEL-3的分子量分别为4.277×10⁶,4.276×10⁶,4.203×10⁶,4.111×10⁶ Da。APEL及其组分对α-淀粉酶和α-葡萄糖苷酶均有显著抑制作用,并能显著延长活化部分凝血活酶时间、凝血酶原时间和凝血酶时间,具有明显的体外降血糖和抗凝血活性。     

Rumen degradation of cell wall polysaccharides in untreated and alkali-treated straws and cereal brans

Rumen degradation of cell wall polysaccharide constituents using the in sacco technique was studied for untreated cereal brans (oats, wheat and barley) and nine technically alkali-treated batches of barley straw. The alkali treatment included four ammonia treatments, two dry sodium hydroxide treatments and three wet sodium hydroxide treatments. On degradation of the cell wall polysaccharides, their main constituents, xylose and glucose, were released at about the same rate for the ammonia treatments and the dry sodium hydroxide treatments. For the wet sodium hydroxide treatments the degradation of the xylose residues was faster than the liberation of the glucose residues. For all treatments the liberation of the arabinose residues was faster than that of the xylose and glucose residues. The reason for the increased digestibility of the cell wall polysaccharides is suggested to be the breaking of ester and hydrogen bonds and the breaking of alkali-labile linkages in lignin, as well as changes in the surface layer of the straw. For the wet sodium hydroxide treatments the larger amount of water present during the alkali treatment processes is suggested to increase the diffusion of the alkali into the straw and to translocate a part of the hemicellulose. On the degradation of wheat and barley bran the liberation of xylose residues was fast whereas xylose and arabinose residues in oat bran were slowly liberated.     

紫苏籽多糖分离纯化及抗肿瘤活性

为了探究紫苏籽多糖（Perilla frutescens seed polysaccharide,PFSP）的抗肿瘤活性,利用传统方法从紫苏籽中提取粗多糖,利用DEAE-52纤维素和葡聚糖G-100柱层析分离纯化获得紫苏籽多糖,并测定该多糖对H22荷瘤小鼠免疫系统的影响。结果表明,紫苏籽粗多糖得率为8.38%,分离纯化后得到3个组分,分别命名为PFSP-1、PFSP-2和PFSP-3。PFSP-1由甘露糖、半乳糖、木糖及阿拉伯糖（物质的量之比为0.01∶0.06∶0.11∶0.81）组成,PFSP-2由甘露糖、木糖及阿拉伯糖（物质的量之比为0.28∶0.28∶0.41）组成,PFSP-3由鼠李糖、葡萄糖醛酸、葡萄糖、半乳糖、木糖及阿拉伯糖（物质的量之比为0.013∶0.024∶0.040∶0.080∶0.120∶0.700）组成;3个组分均具有多糖特征吸收峰,为吡喃环型多聚糖。抗肿瘤实验表明,PFSP-2抑瘤率显著高于PFSP-1和PFSP-3(P<0.05);对PFSP-2进一步研究发现,与模型组相比,PFSP-2可显著降低乳酸脱氢酶、醛缩酶和白细胞介素（interleukin...     

1-甲基环丙烯对硬溶质型桃果实细胞壁多糖降解特性的影响

以硬溶质型桃"晚湖景"为试材,20℃下用1μL/L 1-甲基环丙烯(1-MCP)密闭处理24h,研究1-MCP处理对细胞壁降解过程中多糖组分变化、单糖解离特性以及细胞壁多糖降解相关酶的影响。结果表明:1-MCP处理可以明显抑制硬溶质型桃果实硬度的下降,降低乙烯释放量。1-MCP处理的硬溶质型桃果实中CDTA-2、Na2CO3、KOH和CWM-残渣组分在贮藏过程中下降速率都明显低于未处理果。1-MCP处理能延缓CDTA溶性果胶多糖的解离。同时还抑制了Na2CO3溶性果胶多糖中半乳糖醛酸和鼠李糖构成的主链和阿拉伯糖、半乳糖支链的降解,对半纤维素和纤维素多糖成分降解有一定程度的抑制作用。1-MCP处理可能通过抑制参与细胞壁多糖降解相关酶的活性,从而抑制硬溶质型桃果实细胞壁多糖降解及单糖解离,达到延缓果实软化的目的。     

茶多糖不同提取工艺的比较研究

用醇沉淀法、超滤法和CTAB沉淀法分别制备了茶多糖粗提物,对粗提物的组成进行了分析,并对多糖的清除羟基自由基的能力进行了比较。结果表明,超滤法所得多糖纯度高,且活性也最高,对羟基自由基抑制率可分别比CTAB沉淀法、醇沉淀法提高23.5％、37.1％。通过纸层析和气相色谱分析,三种工艺所提取的茶多糖均由阿拉伯糖、木糖、核糖、葡萄糖、半乳糖及半乳糖醛酸组成,但组成比不同。     

双孢菇菇柄多糖柱层析纯化及单糖组成

以双孢菇菇柄为试材,首先对经超声提取得到的菇柄多糖进行柱层析分离纯化,然后对柱层析得到的多糖组分别采用紫外光谱和柱层析进行纯度分析,并进行红外光谱分析和单糖组成分析。结果表明,脱蛋白双孢菇菇柄多糖经DEAE Sephadex A-25柱层析纯化共得到4个组分,收集其中两个较多的组分(蒸馏水和0.1 mol/L NaCl洗脱组分),经紫外光谱扫描无核酸和蛋白质,经Sephadex G-200柱层析鉴定均为单一峰。所得两种多糖组分经FT-IR红外光谱分析,均含有多糖特征吸收峰,且均为吡喃糖环β-异构体的多糖。菇柄蒸馏水洗脱多糖组分由葡萄糖、半乳糖、甘露糖、木糖、阿拉伯糖组成,0.1 mol/L NaCl洗脱组分是由葡萄糖、半乳糖、甘露糖、木糖、阿拉伯糖、鼠李糖、氨基葡萄糖、氨基半乳糖组成。     

Separation and quantification of component monosaccharides of the tea polysaccharides from Gynostemma pentaphyllum by HPLC with indirect UV detection

A reversed-phase high-performance liquid chromatographic (HPLC) method is described for the simultaneous determination of aldoses and uronic acids. The separation was carried out on a RP-C₁₈ column (4.6 mm i.d. × 250 mm, 5 μm, Venusil, USA) using precolumn derivatization with 1-phenyl-3-methyl-5-pyrazolone (PMP) and UV detection at 250 nm, and the 10 PMP derivatives of mannose, ribose, rhamnose, glucuronic acid, galacturonic acid, glucose, xylose, galactose, arabinose and fucose were baseline separated within 40 min. Furthermore, the described method was applied to the quantitative analysis of component monosaccharides in the water-soluble polysaccharides extracted from Gynostemma pentaphyllum Makino tea and the result showed that the tea polysaccharide was a typical heteropolysaccharide and consisted of mannose, ribose, rhamnose, glucuronic acid, galacturonic acid, glucose, xylose, galactose and arabinose in the molar contents of 16.3, 10.3, 47.1, 5.6, 24.0, 128.4, 25.0, 101.4 and 71.1 μM, respectively. Quantitative recoveries of the component monosaccharides in the tea polysaccharide were in the range of 94.6–108.0% and the RSD values were lower than 4.9%. The results demonstrated that the proposed HPLC method was precise and practical for the analysis of the G. pentaphyllum tea polysaccharide.     

一种新型板栗黄酒多糖的提取分离及其体外免疫活性

以糯米为主要原料、板栗作为一种营养强化成分，按照传统黄酒的发酵工艺，酿制出一种具有丰富营养成分的新型板栗黄酒。通过乙醇分级沉淀法得到板栗黄酒多糖CWP，并使用超滤技术对其进行分离纯化，最终获得2种具有不同多糖分子质量段的板栗黄酒多糖CWP-1(>100 ku)和CWP-2(30～100 ku)。CWP-1和CWP-2主要由8种单糖成分组成，包括半乳糖、甘露糖、葡萄糖、阿拉伯糖、木糖、岩藻糖、半乳糖醛酸和葡萄糖醛酸。红外光谱分析表明，2种多糖均显示出多糖的典型吸收峰，并且CWP-1存在α-和β-糖苷键构型。体外免疫活性实验表明，CWP-1和CWP-2能够显著诱导小鼠巨噬细胞RAW264.7分泌肿瘤坏死因子-α、白细胞介素6和一氧化氮，并且其分泌细胞因子能力高于在相同质量浓度下的脂多糖。本研究结果将为板栗深加工产品开发和新型黄酒中功能性成分的研究和利用提供理论参考。     

鲜食水稻籽粒成熟过程中多糖组成及抗氧化活性分析

以不同成熟时期的鲜食水稻籽粒为原料,采用超声—微波协同萃取法提取多糖,同时利用HPLC法分析了多糖的组成,并通过体外抗氧化试验测定其抗氧化活性。试验结果表明,乳熟后期,鲜食水稻中多糖含量最高,达46.89mg/g;此时的多糖组分主要为甘露糖、鼠李糖、葡萄糖醛酸、半乳糖醛酸、葡萄糖、半乳糖、木糖、阿拉伯糖和岩藻糖等,而完熟期鲜食水稻的多糖组分中并未检出甘露糖和岩藻糖;通过对不同时期鲜食水稻中多糖的抗氧化能力研究发现,乳熟后期的鲜食水稻籽粒中,多糖对ABTS+自由基、DPPH自由基、羟基自由基、超氧阴离子清除率分别为86.18%,56.81%,75.62%,41.69%,均强于其他时期。     

Studies on the free sugars,water-soluble gums and hemicelluloses from barley grains and malts

The free sugars, water-soluble gums, and hemicelluloses have been isolated from four barley grain samples [Hordeum vulgare (L.)], and from their commercial malts. Arabinose, xylose, galactose, glucose, fructose, maltose and sucrose occur as free sugars in the grains and the proportions of these sugars increase on malting. Mannose also occurs as a free sugar in the malts. The water-soluble gums contain arabinose, xylose, lesser amounts of mannose and galactose, and varying amounts of glucose. The aqueous-ethanol supernatants from the hot-water extracts are glucans of low molecular weight. The hemicellulose A polymers contain arabinose, xylose, mannose and glucose. Wide variations in the mannose and glucose contents were observed in these polysaccharides. The hemicellulose B polymers are arabinoxylans, containing lesser amounts of galactose and glucose. The aqueous-ethanol supernatants from the hemicellulose B precipitations contain arabinose, xylose, mannose, galactose and glucose. These polysaccharides differ markedly from those present in the aqueous-ethanol supernatants of the hot-water extracts. All polysaccharides are modified to varying degrees during malting.     

Digestion of cell wall components by dairy heifers fed diets based on alfalfa and chemically treated oat hulls.

Four Holstein heifers were used in a 4 X 4 Latin square design to measure total tract digestion of cell wall components from diets based on alfalfa haylage and alkaline hydrogen peroxide-treated oat hulls. Diets contained 90% forage and 10% concentrate. Treatments were diets containing 90, 70, 50, or 30% alfalfa haylage with treated oat hulls supplying the remainder of the forage portion. Total tract digestion of cell wall-associated uronic acids, arabinose, galactose, mannose, rhamnose, and lignin were not affected by forage source. Digestibilities of cell wall glucose and xylose increased with increasing level of dietary treated oat hulls, reflecting the positive effect of alkaline hydrogen peroxide treatment on cell wall digestion. Cellulose (ADF minus acid detergent lignin) digestibilities were similar to those for cell wall glucose, whereas hemicellulose (NDF minus ADF) digestibilities were similar to those for cell wall arabinose plus xylose. Low digestibilities of alfalfa cell wall xylose indicate that some cell wall structure inhibits the degradation of alfalfa xylans. Low degradabilities of core lignin, esterified p-coumaric acid, and esterified acetyl groups suggest that these components may be involved primarily in depressing fermentation of cell wall polysaccharides.