毛细管电泳法测定芦根和蒲黄多糖中的单糖组分(英文)

本文建立了用毛细管电泳测定中草药芦根和蒲黄多糖中的单糖组分的方法。提取的单糖经α-萘胺衍生,用二极管阵列检测器在214nm处检测。五种单糖鼠李糖(Rha),木糖(Xyl),半乳糖(Gal),葡萄糖(Glu),阿拉伯糖(Ara)能够同时测定,分离良好。毛细管电泳的测定条件如下,50 mM硼酸钠缓冲液;pH10.5;工作电压为25kV;温度为25℃。标准曲线的线性范围为5 mg/L-200 mg/L,每种单糖的最低检测线均为2.5 mg/L。迁移时间和峰面积的相对标准偏差分别小于1.46%,6.21%。实验结果显示,芦根多糖重各单糖组分的摩尔比如下:木糖∶葡萄糖∶阿拉伯糖∶半乳糖为1.00∶59.03∶2.09∶1.96;蒲黄多糖中各单组分的摩尔比如下:鼠李糖∶木糖∶葡萄糖∶阿拉伯糖∶半乳糖为1.71∶1.00∶9.30∶6.88∶1.24。     

浒苔多糖的分级纯化及保湿活性研究

用DEAE-Sepharose FF离子交换凝胶层析柱对浒苔(Enteromorpha)多糖进行分级纯化,利用HPLC色谱分析了单糖组成,并以甘油和透明质酸为对照,研究其吸湿和保湿活性。结果表明,浒苔多糖主要为葡萄糖醛酸-鼠李糖-木糖-甘露糖聚合物;浒苔多糖主要由两种组分组成,其一组分木糖和鼠李糖含量摩尔比为1.11:1,另一组分木糖和鼠李糖含量比为1:3.28;浒苔多糖具有与透明质酸相似的吸湿和保湿能力。     

龙须菜(Gracilaria lemaneiformis)多糖的分离纯化及其组成研究

以龙须菜粗多糖为原料,利用离子交换层析分离纯化,紫外光谱、醋酸纤维薄膜电泳和凝胶过滤层析法鉴定多糖的纯度及其分子量,同时进一步采用气相色谱分析龙须菜多糖的单糖组成.结果表明,利用蒸馏水、0.3mol/L NaCl和1.2mol/L NaCl溶液分步洗脱,DEAE-Sephadex A-25离子交换层析纯化得到龙须菜多糖的三个组分G1、G2和G3,得率分别为3.61%、69.26%、18.70%:G1、G2和G3均显示为单一电泳谱带和单一洗脱峰,无蛋白、核酸杂质,分子量依次为21993、48460、61031U;G2和G3的单糖组成主要为半乳糖、3.6-内醚半乳糖及少量葡萄糖;G2糖链通过1→2键与1→6键方式连接,存在非还原末端基及连三羟基,G3糖链则通过1→3键与1→2键方式连接.     

4种海蕴多糖的提取分离及理化性质

以海蕴(Nemacystus decipiens)为研究对象,经85%乙醇脱脂后,通过热水和2%碳酸钠提取并结合乙醇分级法获得了4个多糖组分(NW1~2和NA1~2)。运用HPIC、HPGPC及红外光谱(IR)等方法对其单糖组成、相对分子量以及基本结构特征进行了比较。结果表明,NA1是褐藻胶,经1H-NMR分析,其M/G为0.34。NA2及NW1~2均属于岩藻聚糖硫酸酯,且岩藻糖含量均达80%以上。NW1和NW2中岩藻糖(Fuc)、葡萄糖(Glc)、半乳糖(Gal)、木糖(Xyl)和鼠李糖(Rha)摩尔比分别为100∶5∶3∶16∶1和100∶5∶4∶4∶1。NA2中不含葡萄糖,其Fuc、Gal、Xyl和Rha摩尔比为100∶13∶1∶2。多糖NW1~2和NA1~2的相对分子量分别为939.8,1001,223.6和402.3 kD。     

梅花参中多糖提取工艺及含量测定的研究

采用正交设计L9（34）,研究梅花参（Thelenota ananas）中多糖的提取工艺,探讨并建立了测定海参多糖含量的方法.结果表明：最佳提取条件为1g样品加40mL 3%KOH溶液,40～45℃超声2h;且提取温度为首要的影响因素;产物经二次乙醇纯化的结果较好,粗多糖得率20.25%.多糖经单糖组成分析,岩藻糖含量达到70%以上.测得梅花参中总糖含量8.79%;糖醛酸含量0.95%.该实验操作简单,稳定可靠,重现性好,准确度高,为测定其它海参中多糖的含量提供了可靠的依据.     

厚藤共生真菌(Fusarium oxysporum Y24-2)胞外多糖的化学组成和结构研究

从厚藤共生真菌(Fusarium oxysporum Y24-2)发酵液中提取得到胞外多糖,以Q-Sepharose Fast Flow阴离子交换柱和Superdex S-75凝胶色谱柱对其进行分离纯化,得到纯度高的多糖组分F1S。采用HPGPC和HPLC对F1S的分子量及单糖组成进行分析,结果表明F1S主要由Man、Glc和Gal组成,其摩尔比为1.0∶1.9∶2.1,分子量为37.3kD。F1S的GC-MS和1D,2D-NMR分析表明,该多糖是以→6)-β-Galf(1→为主链,以→2)-α-Glcp(1→,α-Glcp(1→和β-Manp(1→为支链的杂多糖,支链取代于→6)-β-Galf(1→的O-2位。该厚藤共生真菌的胞外多糖为富含呋喃型半乳糖的结构新颖的具有多分支结构的中性杂聚多糖。     

海绵内生真菌胞外多糖理化性质及自由基清除活性研究

从1株海绵内生真菌(Alternaria sp.)链格孢菌的发酵液中提取胞外多糖,对其进行Q Sepharose FF离子交换色谱分离纯化,得到2个组分JJY-W和JJY-S。运用各种化学方法及波谱方法对JJY-W和JJY-S的理化性质及结构进行分析,并对多糖清除自由基活性进行了评价。结果表明,JJY-W和JJY-S的分子量分别为1.4KD和1.8KD;JJY-W以半乳糖、葡萄糖为主,含有少量甘露糖,摩尔比为:甘露糖∶葡萄糖∶半乳糖=1∶2.5∶11。JJY-S以甘露糖、葡萄糖为主,含有少量半乳糖,摩尔比为:甘露糖∶葡萄糖∶半乳糖=3∶2∶1。JJY-W总糖含量为46.3%,未检测出糖醛酸,蛋白含量为2%。JJY-S总糖含量为52%,糖醛酸含量为6.1%,蛋白含量为14%。活性分析表明,2种多糖均具有一定的体外抗氧化活性,并随着浓度的增加清除自由基能力均增强,JJY-W清除DPPH自由基的活性要强于JJY-S,而JJY-S清除OH.自由基的活性要强于JJY-W。     

自养小球藻硫酸多糖的红外光谱分析

利用二乙胺基乙基琼脂糖凝胶柱层析纯化自养小球藻(Chlorella autotropica)粗多糖,得到3个硫酸多糖组分(A,B,C);利用高效液相色谱仪和熔点仪测定了3个多糖的分子质量和熔点;利用BaCl2浊度法测定了硫酸多糖中硫酸根的质量分数;利用傅立叶变换红外光谱分析了海水自养小球藻3个硫酸多糖组分的环状结构、半缩醛羟基构型及取代基类型。结果显示,3个硫酸多糖组分的硫酸根的质量分数分别为0.77%,8.46%,2.69%,3个组分均含有硫酸基、酰氨基,葡萄糖是3个硫酸多糖组分的主要组成单糖;3个硫酸多糖组分组成单糖的环构型均是吡喃环,成苷的半缩醛羟基构型有一定差异,A是α构型,B、C是α、β两种半缩醛羟基构型并存。为深入研究自养小球藻硫酸多糖的结构及抗癌活性提供基础。     

南极菌Psychrobacter sp.B-3胞外多糖化学特征及其对番茄抗氧化酶的诱导作用研究

以B-3多糖诱导番茄叶,采用试剂盒测定其对番茄叶抗氧化酶活性的影响,并对南极茵B-3胞外糖的分子量、单糖组成等化学特征进行了初步分析.结果表明,不同质量浓度的B-3在诱导不同时间后,均能提高番茄各抗氧化酶的活性.0.5％的B-3多糖诱导24 h过氧化氢酶(CAT)活性最高;0.5％的B-3多糖作用48h对超氧化物歧化酶...     

4种不同来源浒苔中多糖的提取分离及理化性质

为探讨不同季节以及不同地点采集的浒苔其多糖理化性质差异,采用水提醇沉法对黄海海域青岛沿海春(原料1)秋(原料3)两季采集的浒苔、2008年夏季爆发浒苔(原料2)以及东海海域福建沿海采集的浒苔(原料4)的多糖进行了提取,得到了4种冷水提取多糖PC1-4和4种热水提取多糖PH1-4。分别对8种浒苔多糖利用Q-Sepharose Fast Flow离子交换色谱法纯化后得到了12个组分。对各组分的总糖含量、糖醛酸含量、硫酸根含量以及蛋白含量进行了测定,利用气相色谱法测定了单糖组成,结果显示不同季节不同海域采集样品的化学组成及单糖组成不同。     

冬季中国东海中溶解碳水化合物浓度分布

采集了中国东海(2009-12~2010-01)35个站位的海水样品,其中包括6个站位的垂直断面。用TPTZ方法测定了溶解态的单糖(MCHO)、多糖(PCHO)和总糖(TCHO)的浓度,对其水平分布和PN断面分布进行了研究。结果表明,受到长江冲淡水和黑潮水的影响,表层海水中MCHO、PCHO和TCHO浓度的水平分布表现出由近岸向外海递减的分布趋势。PN断面由于受冬季东北季风的影响,水体混合强烈,使得MCHO、PCHO和TCHO浓度在垂直方向上分层无明显规律。对长江口外3个断面海水中TCHO浓度与环境因子做了相关性研究。结果表明,TCHO浓度与盐度和温度呈显著的线性负相关,与Chl-a浓度呈线性正相关。     

Dynamics of carbohydrates in the Norwegian Sea inferred from monthly profiles collected during 3 years at 66°N, 2°E

Water samples were collected monthly for 3 years at 66°N, 2°E in the Norwegian Sea, 250 nautical miles off the Norwegian coast. Concentrations of mono- and polysaccharides were measured with the 2,4,6-tripyridyl-s-triazine (TPTZ) spectroscopic method. Total dissolved carbohydrates varied from 3.4 to 28.2 μM C of all samples and the ratio of carbohydrate to dissolved organic C (DOC) varied from an average of 14% at 0–25 m depth to 11% at 800–2000 m depth. This indicates that dissolved carbohydrates were a significant constituent of DOC in the Norwegian Sea. Polysaccharides varied from 0.4 to 21.5 μM C and monosaccharides from 0.7 to 11.7 μM C at all depths. The level of monosaccharides was relatively constant at 2.8–3.2 μM C below the euphotic zone, whereas polysaccharides showed more varying concentrations. Dissolved carbohydrates accumulated during the productive season, reaching maximum concentrations during summer although interannual differences were observed. A significant positive correlation between Chl a and soluble carbohydrate was found in one growing season with nutrient analyses. Average values for total carbohydrates were highest in the surface – 0 to 25 m – with 13.3 μM C and decreased to 8.4 μM C at 800–2000 m depth. The ratio of monosaccharides to polysaccharides exhibited a marked seasonal variation, increased from January to a maximum in June of 1.1, and declined to 0.5 in July.     

南极普里兹湾沉积物中的糖类分布及意义

利用中国第18和21次南极考察获取的南极普里兹湾沉积物样品,分析了其中的有机碳和糖类物质的含量及组成,结果表明糖类和有机碳的分布受上层水体的初级生产、地形条件和水体垂直稳定度等多种因素的控制。表层沉积物中糖类物质的平均含量为3.03 mg/g,最高值为5.60mg/g,出现在湾内的毗邻陆架区。沉积物有机碳含量与表层海水叶绿素a具有良好的相关性,能够反映上层水体初级生产的变化。单糖组分的研究可以判定其生源母质,沉积物中有机质的来源主要是海洋上层生物。糖类是易被降解利用的有机质,通过糖类物质中六碳糖的比重及其垂直分布的变化可以判断出不同站位沉积速率的相对快慢。     

Distributions of carbohydrates, including uronic acids, in estuarine waters of Galveston Bay

The concentrations of carbohydrates, including uronic acids, in dissolved (≤0.45μm) and colloidal (1 kDa—0.45 μm) phases were measured in estuarine waters of Galveston Bay, TX, in order to study their role in heavy metal detoxification. The concentrations of dissolved monosaccharides (MCHO) in Galveston Bay ranged from 13 to 62 μM-C, and those of dissolved polysaccharides (PCHO) ranged from 10 to 42 μM-C. On average, MCHO and PCHO contributed about 11% and 7% to dissolved organic carbon (DOC), respectively. The colloidal carbohydrates (CCHO) in Galveston Bay varied from 7 to 54 μM-C, and accounted for 9% to 24% of the colloidal organic carbon (COC), with an average value of 17%, suggesting that CCHO is abundant in the high molecular weight (HMW) fraction of DOC. The concentration of CCHO is generally significantly higher than that of PCHO. This result is attributed to entrainment of low molecular weight (LMW) carbohydrates into the retentate fraction during ultrafiltration. The concentration of total dissolved uronic acids (DUA) in the same samples varied from 1.0 to 8.3 μM-C, with an average value of 6.1 μM-C, while the colloidal uronic acids (CUA) ranged from 0.8 to 6.4 μM-C, with an average value of 4.8 μM-C. The concentrations of DUA are higher than the previously reported values in coastal waters. Furthermore, CUA represent a dominant component of DUA in Galveston Bay waters. More importantly, significant correlations of PCHO and DUA to dissolved Cu concentrations (≤0.45 μm) were found, suggesting that acid polysaccharides were produced in response to trace metal stressors.     

An inventory of factors that affect polysaccharide production by Phaeocystis globosa

Phaeocystis material contains polysaccharides that are built from at least eight different monosaccharides. Differences have been reported between the carbohydrate composition of different Phaeocystis species, and also between samples taken from Phaeocystis globosa blooms in different areas. In order to elucidate factors that could play a role in determining variation in carbohydrate composition and production, a number of Phaeocystis globosa strains were studied under laboratory conditions. Although there was a clear distinction of a northern and a southern cluster in the Phaeocystis globosa strains based on RAPD analysis, the differences in the composition of the mucopolysaccharides were relatively small. The contribution of glucose, however, ranged from 7–85% of total sugars. A strain that was cultured in seawaters of diverse origin produced polysaccharides of a different composition, suggesting the effect of environmental factors. The presence of bacteria affected neither the amount, nor the composition of the carbohydrates that were produced by Phaeocystis globosa. Glucose is part of both the intracellular polysaccharide pool and of the mucopolysaccharides in the colony matrix. Using specific digestion of the intracellular chrysolaminaran by laminarinase, the distribution of polysaccharides over different pools could be assessed. During growth of an axenic, mucus-producing strain, the portion of glucose present as chrysolaminaran appeared to increase. The polyglucose that was not digested by laminarinase remains unidentified. This study shows that environmental factors rather than strain differences determine differences in the sugar composition of Phaeocystis globosa, especially with respect to the glucose content of the material. A difference in the contribution of glucose could be correlated to the portion of cells in the culture that are not in the colonies. Our study emphasises that for studying polysaccharide dynamics in Phaeocystis globosa it is important to be able to discriminate between the different polysaccharide pools. Preliminary results of an enzymatic approach were promising     

几种海藻粗多糖的理化性质及结构特征分析

主要采用傅里叶红外光谱(fourier transform infrared spectroscopy, FTIR)、气相色谱(gas chromatography, GC)和凝胶渗透色谱(gel permeation chromatography, GPC)方法对5种海藻粗多糖的单糖组成、官能团、分子量进行了初步分析。结果显示5种海藻粗多糖均含有木糖和岩藻糖, 其中南方团扇藻多糖含有较为丰富的山梨糖, 属于大分子量均一性多糖, 而孔石莼多糖分子量分布较宽, 均一性较差。红外光谱分析结果表明, 5种粗多糖均为酸性多糖, 棒叶蕨藻、琼枝麒麟菜和匍枝马尾藻多糖具有明显的硫酸基团吸收峰, 琼枝麒麟菜多糖存在呋喃糖和吡喃糖, 并且吡喃糖苷键可能存在β和α两种类型。明胶氯化钡实验结果指出南方团扇藻多糖的硫酸基团含量为38.43%(±2.38%), 明显高于其他几种海藻多糖, 而棒叶蕨藻多糖的硫酸基团含量最低, 仅为8.92%(±1.04%)。     

雨生红球藻多糖的提取分离及理化性质研究

将雨生红球藻(Haematococcus pluvialis)经乙醇/乙酸乙酯(1:2,V/V)脱脂,通过冷水、热水和5%碳酸钠提取,得到3种粗多糖(HPC、HPH和HPA),通过Q-Sepharose FF阴离子交换柱进一步分级,从HPC中得到HPC1、HPC2和HPC3共3种多糖组分。运用高效液相色谱法(HPLC)、红外光谱(IR)等方法对其单糖组成、相对分子质量和基本结构特征进行分析比较。结果表明,各组分单糖组成复杂,主要含有半乳糖(Gal)、甘露糖(Man)、阿拉伯糖(Ara)、葡萄糖(Glc)、葡萄糖醛酸(GlcUA)、半乳糖醛酸(GalUA)、木糖(Xyl)、鼠李糖(Rha)、岩藻糖(Fuc)、葡萄糖胺(GlcN)和半乳糖胺(GalN),其中,Gal含量最高,大于20%。多糖HPC1、HPC2和HPC3的相对峰位分子质量分别为502.6,373.2,577.5和300.5 ku。     

两种多糖作为迟缓爱德华氏菌(Edwardsiella tarda)灭活疫苗佐剂对大菱鲆(Scophthalmus maximus)的免疫保护效果

采用黄芪多糖、葡聚糖作为免疫佐剂,与迟缓爱德华氏菌灭活疫苗配伍后注射免疫大菱鲆,测定免疫28d后血清中溶菌酶活力、SOD活力、抗体效价和各免疫组的相对保护率(RPS)。结果表明,添加多糖免疫佐剂能提高疫苗免疫的大菱鲆的各免疫指标,添加佐剂的免疫组的血清溶菌酶活力、SOD活力和血清效价比单纯的疫苗免疫组显著提高(P<0.05),2.5mg/ml黄芪多糖混合疫苗免疫组和5mg/ml葡聚糖混合疫苗免疫组的相对保护率最高,分别达(78.7±1.3)%和(64.0±8.9)%,且溶菌酶活力、SOD活力及血清效价等指标较其他各组有提高。