桑黄菌丝体多糖的分离纯化及理化性质分析

研究桑黄粗多糖的分离和纯化工艺,并对多糖组分进行理化性质分析。结果表明：经DEAE-52纤维素离子交换层析和Sephadex G-100凝胶过滤层析得到两个组分P-47000和P-8700;经高效液相色谱分析证明,两组分均为纯品,且不含蛋白质、核酸,为非淀粉类多糖,分子质量分别为4.74×104D和8.71×103D,均由鼠李糖、阿拉伯糖、木糖、葡萄糖、半乳糖组成,P-47000中各单糖物质的量比为3.47:1.99:1:63.27:13.44,P-8700中各单糖物质的量比为10.46:1:1.03:182.75:30.94。     

Structural investigation of a novel water-soluble heteropolysaccharide from the fruiting bodies of Phellinus baumii Pilát

A novel water-soluble heteropolysaccharide termed PBF2 was isolated from the fruiting bodies of Phellinus baumii Pilát using hot water extraction and purified on Sephacryl S-1000. PBF2 was shown to be composed of l-fucose, d-mannose and d-glucose. Its structural characteristics were further investigated by FT-IR spectroscopy, sugar analysis, methylation analysis and NMR spectroscopy. Based on the data obtained, PBF2 was found to be a heteropolysaccharide containing a β-(1 → 6)-d-glucopyranose backbone with a fucosyl unit on O-3 of the 3,6-di-O-substituted-d-glucosyl units and it also contained a minor (1 → 3, 6)-β-d-mannose residue and terminal glucose residues.     

桑黄胞内多糖免疫及抗氧化活性研究

通过腹腔注射环磷酰胺建立免疫抑制小鼠动物模型,考察桑黄粗多糖(SH)、纯化多糖组分(P-47000、P-8700)对增强免疫及抗氧化水平的变化情况,明确不同纯度多糖组分生物活性间的差异。结果表明：SH、P-47000、P-8700对环磷酰胺诱导的免疫功能低下小鼠具有不同程度的免疫促进作用,其顺序为：P-8700＞SH＞P-47000;P-47000的还原能力低于SH和P-8700,SH和P-8700还原能力相差不大;P-47000几乎对超氧阴离子自由基(O2－ ·)没有清除效果,P-8700和SH的清除率比较低,且P-8700略大于SH;P-8700对羟自由基( ·OH)具有较好的清除效果,清除率达到53.421%,且P-8700的清除率高于SH和P-47000,P-47000清除率最低;不同纯度多糖对DPPH自由基的清除效果均较高,SH达到93.221%;P-8700清除效果最低,为66.435%。     

响应面法优化桑黄产胞内多糖液体发酵培养基

利用响应面分析法对桑黄菌丝体生物量及产胞内多糖的液体发酵培养基进行优化,研究碳源、氮源、无机盐对桑黄菌丝生物量、胞内多糖含量及产量的影响。在单因素筛选试验的基础上,利用Box-Benhnken设计和响应面分析法对碳源、氮源和无机盐水平进行分析。结果表明,桑黄产胞内多糖的液体发酵培养基最佳组合为:玉米粉3.9%、麸皮2.2%、KH2PO4 0.20%、MgSO4 0.10%,在此条件下的验证实验表明,胞内多糖产量可达233.107mg/L。     

松木层孔菌多糖PP60-S1分离、结构表征及抗氧化活性研究

松木层孔菌子实体水提物经乙醇分级醇沉和DEAE-Sepharose Fast Flow离子柱层析及Sephacryl S-300凝胶柱层析纯化,获得均一多糖PP60-S1,其分子量为3.56×104u,是由葡萄糖、甘露糖和半乳糖组成的杂多糖,单糖摩尔比为9.24∶1.00∶0.76,甲基化分析PP60-S1主要由1-Glc、1,3-Glc、1,6-Glc、1,3-Gal、1,6-Gal、1,3,6-Man 6种糖苷键连接方式,主链主要由1,6-Glc构成。PP60-S1的体外抗氧化活性研究表明其具有一定的DPPH自由基和ABTS+自由基清除活性以及FRAP总抗氧化能力。      

桑黄子实体两种新多糖的分离纯化与结构研究

目的从桑黄子实体中分离具有抗肿瘤活性的多糖成分,并进行结构分析。方法热水浸提法提取桑黄子实体中的粗多糖,经醇沉、冷冻干燥、离子交换层析、凝胶层析和超滤,得到纯度较高的多糖,通过HPLC、IR、1H-NMR、13C-NMR、高碘酸氧化及Smith降解确定2种多糖的相对分子质量、组成及结构。结果分离纯化得到相对分子质量分别为14.2×103,22.2×103的多糖PL-A及蛋白聚糖PL-B,两者都是结构复杂的中性杂多糖。PL-B中,糖占71%,蛋白质占7%;PL-A,PL-B均含大量葡萄糖及少量甘露糖,PL-B中还有部分鼠李糖;PL-B所含氨基酸包括缬氨酸、赖氨酸、天冬氨酸、甘氨酸。结论多糖PL-A与蛋白聚糖PL-B为首次从桑黄子实体中提取到的多糖成分。     

Isolation,Purification, and Structural Features of a Polysaccharide from Phellinus linteus and Its Hypoglycemic Effect in Alloxan‐Induced Diabetic Mice

Phellinus linteus is a medicinal mushroom that has been used in Oriental countries for centuries for its antitumor, antioxidant, immunomodulatory, and biological activity on hyperglycemia. A water‐soluble crude polysaccharide was extracted using hot water from P. linteus mycelia grown under submerged culture. An orthogonal experiment was used to optimize the extraction conditions of P. linteus mycelia polysaccharides (PLP). The crude polysaccharide was purified using DEAE Sephadex A‐50 and Sephadex G‐200 chromatography. Fourier transform infrared (FT‐IR) spectroscopy and nuclear magnetic resonance (¹H NMR) spectroscopy were used to investigate the structure of the purified P. linteus polysaccharide (PLP‐I), revealing that it was mainly a branched‐type glycan with both α‐ and β‐linkages and a pyranoid sugar ring conformation. PLP orally administered at 100 mg/kg body weight/d could significantly reduce the blood glucose level by 35.60% in alloxan‐induced diabetic mice. The results of an oral glucose tolerance test (OGTT) revealed that PLP had an effect on glucose disposal after 28 d of treatment. The result revealed that PLP from a submerged culture of P. linteus mycelia possessed potent hypoglycemic properties. The polysaccharide may be useful as a functional food additive and a hypoglycemic agent.     

桑黄多糖提取过程模型的建立与动力学分析

基于超声波辅助提取中药材中化学成分的非稳态扩散过程,根据Fick第二定律建立桑黄子实体多糖的超声波协同纤维素酶法提取过程的动力学模型。以桑黄子实体为原料,分别对桑黄多糖的热水提取和超声法协同纤维素酶提取过程建立动力学模型,并对模型进行试验验证,求解相应的动力学参数;得到速率常数、活化能、相对萃余率等一系列动力学参数。研究结果显示试验数据与动力学模型计算值吻合良好,可为桑黄多糖提取工艺放大和深入理论研究提供一定的依据。     

南瓜皮多糖PP1的提纯、组分分析及原子力显微镜技术观测研究

采用热水浸提法及十六烷基三甲基溴化铵法(CTAB法)提取南瓜皮多糖PP1,紫外分光光度法和高效液相色谱法检测纯度,红外光谱分析其结构及气相色谱测定南瓜皮多糖PP1的单糖组分;通过乙醇固定、Ni2+搭桥固定两种制样方法,原子力显微镜(AFM)观测南瓜皮多糖PP1微观形貌。结果表明：热水浸提及CTAB法分离纯化南瓜皮多糖PP1的纯度较高,不含核酸及蛋白质,红外光谱分析证实南瓜皮多糖PP1具有糖类物质的特征吸收峰,存在吡喃环,由果糖、阿拉伯糖、葡萄糖和半乳糖组成,物质的量比为0.92:2.76:5.52:2.24,AFM观测其结构呈单链形排列,无分支状结构,采用乙醇固定法得到的多糖结构形态较细长,Ni2+搭桥固定法得到的多糖结构形态相对短粗。     

桑黄子实体多糖、黄酮和多酚含量与抗氧化活性相关性

探讨子实体抗氧化的物质基础。通过提取和萃取得到7种组分:水提物(WE)、醇提物(AE)、粗多糖(CP)、醇沉上清干物(RL)和二氯甲烷(MCE)、乙酸乙酯(EAE)、正丁醇(n BE)萃取物,采用FRAP值法测定体外抗氧化能力,分光光度法测定多糖、黄酮和多酚含量。结果表明:抗氧化活性物存在于醇提物中,抗氧化活性和多酚、黄酮含量有关,并呈线性正相关(R2为0.9977和0.9950),多糖在抗氧化中所起作用小,酚类、黄酮类为其主要抗氧化活性物质。      

小分子苦瓜多糖MCPⅡa的纯化及结构分析

利用酶解结合水溶醇沉提取水溶性的苦瓜多糖（Momordica charantia polysaccharide,MCP）,经Sevag法除蛋白、DEAE-纤维素离子交换、Sephadex G-100凝胶过滤获得活性多糖MCPⅡa。由高效凝胶渗透色谱检测可知MCPⅡa为均一多糖,平均分子质量为13.0 kD,单糖组分分析显示其单糖组成为鼠李糖、半乳糖醛酸、半乳糖、木糖和阿拉伯糖（各组分物质的量比为12∶3.05∶19.89∶5.95∶56）。傅里叶红外光谱、核磁共振及刚果红实验发现其存在C1、C2、C3、C5连接,在水溶液中具有稳定的β-三股螺旋构象,扫描电镜下显示其为菱形晶体颗粒。     

Structural investigation of a novel fucoglucogalactan isolated from the fruiting bodies of the fungus Hericium erinaceus

A new heteropolysaccharide with a molecular weight of 1.94 × 10⁴ Da, HEPF1, was isolated from the fruiting bodies of Hericium erinaceus. It is composed of fucose, galactose and glucose in the ratio of 1:4:1, as well as a minor proportion of 3-O-methyl rhamnose. Sugar analyses, methylation analysis, together with ¹H and ¹³C NMR spectroscopy established that HEPF1 has a (1 → 6)-linked α-d-galactopyranosyl backbone with branches that are composed of fucose attached to O-2; it also contains 6-O-substituted-β-d-oligoglucosyl units and a minor terminal 3-O-methyl rhamnose residue.     

鹿蹄橐吾多糖LW21的分离纯化及其结构分析

目的:确定鹿蹄橐吾多糖LW21的结构,为探讨鹿蹄橐吾多糖的药理活性,合理利用鹿蹄橐吾这种植物资源提供依据。方法:水提取醇沉获得鹿蹄橐吾水溶性粗多糖LW,经酸性乙醇分级和DEAE-SephdexA-25纯化得多糖LW21。纸层析、醋酸纤维薄膜电泳和Sepharose CL-4B柱层析进行纯度鉴定,LW21结构分析采用高碘酸氧化、Smith降解、甲基化分析及IR、NMR、GC和GC-MS等方法。结果表明:LW21为均一多糖,相对分子质量约为1.1×106。其单糖组成为鼠李糖(Rha)、阿拉伯糖(Ara)、甘露糖(Man)、葡萄糖(Glc)、半乳糖(Gal),物质的量比为7.4:11.9:25.7:40.0:14.9。多糖LW21为有分枝结构,主链由Glc和Man构成,其中其中Man主要以β(1→2)和β(1→6)糖苷键连接,β(1→2)糖苷键在3-O处和6-O处有分枝,β(1→6)糖苷键在2-O和4-O处有分枝,Glc也主要以β(1→2)及β(1→6)糖苷键连接,β(1→2)糖苷键在6-O处有分枝,β(1→6)糖苷键连接,在2-O处有分枝。分子支链由Ara、Rha、Gal构成。末端残基为Gal、Ara、Rha、Glc。结论:鹿蹄橐吾多糖LW2 1是一新结构多糖,为首次从鹿蹄橐吾中分离得到。     

灵芝多糖的分离纯化及结构鉴定

运用DEAE-Sephadex A25离子色谱和Sepharose CL-6B凝胶色谱分离纯化得到一种灵芝多糖组分GLPS1a。高效液相凝胶渗透色谱法(high-performance gel-permeation chromatography,HPGPC)法测得其呈单一峰,重均分子质量为1.8×105D。GLPS1a经单糖组成分析、红外、核磁共振等手段分析,结果表明单糖组成为阿拉伯糖、半乳糖、葡萄糖和木聚糖,物质的量比率为4:2:10:1。GLPS1a具有一条以β→(1,3)位键合的吡喃葡萄糖主链,同时存在β→(1,3)阿拉伯糖、β-D-(1,4)半乳糖、α-D-(1,2)木糖和α-D-(1,6)葡萄糖的分支残基。     

阿魏菇子实体多糖的结构及其抗氧化活性研究

以从阿魏菇子实体中分离纯化得到的多糖-阿魏菇多糖（Pleurotus ferulae Lenzi polysaccharides,PFLPs）作为研究对象,对其化学结构、链构象以及抗氧化活性进行了研究。实验结果表明,PFLPs含有鼠李糖、木糖、甘露糖、葡萄糖和半乳糖五种单糖,摩尔比为1∶1.26∶0.85∶12.40∶4.31,并且糖链内具有β-D-葡聚糖结构。链构象分析表明PFLPs是一种具有三股螺旋结构的多糖。体外抗氧化活性实验表明,PFLPs具有较高的DPPH自由基、ABTS⁺自由基和羟自由基清除活性,适度的超氧阴离子自由基清除活性,还原力活性的和Fe2+的螯合活性。结果表明,PFLPs是一种很有潜力的天然抗氧化剂。      

杏鲍菇碱溶性多糖的酶法提取及其结构的初步分析

以杏鲍菇子实体为原料提取并分离纯化,得到碱溶性多糖组分,通过采用正交实验得到了提取碱溶性多糖的最佳方案。该多糖经层析方法和Smith降解等反应,证实其为单一组分,该组分多糖由葡萄糖为单一糖基组成;并证实了该多糖具有β(1→3)及β(l→6)连接的糖苷键。     

板栗种仁多糖的提取纯化及体外抗肿瘤活性筛选

以燕山地区板栗为原料,利用加压溶剂萃取法提取板栗种仁粗多糖,以多糖得率为指标,通过单因素结合响应面法优化提取工艺条件。将最优提取条件下得到的粗多糖经除蛋白、脱色素、透析后得到纯化多糖,继而借助制备型高效体积排阻色谱得到板栗多糖的高分子量组分(YCP-H),分析了其形貌特点、结构特征和单糖组成,并针对8种人体肿瘤细胞模型进行了体外抗肿瘤活性筛选。结果表明,高压溶剂萃取法提取板栗种仁粗多糖的最佳提取条件为:当投料量为10 g,静态提取次数为3次时,设定温度60 ℃、时间9 min、压力6 MPa,多糖得率可达19.78%±0.27%。组分YCP-H是具有β-吡喃构型的酸性多糖,重均分子量范围为217~5900 kDa,在扫描电镜下呈现纤维状的微观形貌,单糖组成包含阿拉伯糖、半乳糖、葡萄糖、木糖、甘露糖和岩藻糖。YCP-H对人肝癌细胞HepG2的增殖具有显著(P<0.05)的抑制作用,其半数抑制浓度可达0.08 μg/mL。此外,YCP-H对人结肠癌细胞HCT-116和肺癌细胞A-549的增殖也有较强的抑制作用。     

Purification,composition analysis and antioxidant activity of a polysaccharide from the fruiting bodies of Ganoderma atrum

A water-soluble protein-bound polysaccharide was extracted from the fruiting bodies of Ganoderma atrum and isolated by gel-filtration chromatography. Its primary structural features and molecular weight were characterized by infrared spectrometry, gas chromatography, size exclusion chromatography, amino acid analyzer and high-performance liquid chromatography (HPLC). The data obtained indicated that the glycoprotein contains 10.1% of protein and 17 general amino acids and it is rich in glutamic acid, asparagic acid, alanine, glycine, threonine, and serine. It was mainly composed of mannose, galactose and glucose in a molar ratio of 1:1.28:4.91, with an average molecular weight of about 1013 kDa. The existence of an O-glycosidic linkage in PSG-1 (polysaccharide1) was demonstrated by a β-elimination reaction. The antioxidant activity of the purified polysaccharides was evaluated in vitro by 1,1-diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging assay, self-oxidation of 1,2,3-phentriol assay. Those various antioxidant activities were compared to standard antioxidants vitamin C and BHT. It was found that the scavenging effects of the purified polysaccharides increased with measuring concentration. The results indicated that the purified polysaccharides showed strong DPPH free radical and superoxide anion radical scavenging activities. This study suggested that the purified polysaccharides could potentially be used as natural antioxidants.