灵芝孢子粉水溶性多糖的分离、纯化及结构研究

灵芝孢子粉的热水提取液经醇析,脱脂,去单寡糖后由SepharoseCL 6B柱层析纯化,所得多糖SGL Ⅱ2经高效液相方法鉴定纯度为单一级分,相对分子质量为5 .37×10 4。再经部分酸水解、高碘酸氧化、Smith降解、甲基化分析及IR、GC、GC MS和13 CNMR等方法确定其结构。结果表明多糖SGL Ⅱ 2由葡萄糖和半乳糖组成,为少分支结构,由1→3连接和1→6连接的葡萄糖构成主链,部分1→6连接葡萄糖在3位或4位有分支,侧链为1→4连接的半乳糖,分支末端残基为葡萄糖。     

猴头菌子实体和菌丝体多糖的分离纯化与理化特征的比较

人工栽培的猴头菌子实体和固体培养的菌丝体分别经热水提取,浓缩、醇析后得子实体粗多糖（HFP）和菌丝体粗多糖（HMP）,HFP的得率和多糖含量均高于HMP;两者再经透析、Sevag法脱蛋白、乙醇分级沉淀、SephadexG-100柱层析纯化,得子实体纯多糖hfp-1和菌丝体纯多糖hmp-2,经HPLC检测hfp-1和hmp-2为单一均匀多糖,气相色谱分析表明hfp-1的单糖组成为阿拉伯糖、甘露糖、半乳糖和葡萄糖,摩尔比为：0.12:0.04:1.00:0.71;hmp-2的单糖组成为阿拉伯糖、木糖、甘露糖、半乳糖和葡萄糖,磨尔比为：0.25:0.41:0.31:1.00:0.29。紫外光谱分析表明组成中不含核酸和蛋白质,红外光谱分析二者均有多糖特征吸收峰,hfp-1有β-型连接的吸收峰,hmp-2无明显的β-型连接的吸收峰;经HPLC进行分子量测定,hfp-1的分子量为54000,hmp-2的分子量为68000,猴头菌子实体多糖和菌丝体多糖在化学组成上有一定的差异。     

羊肚菌多糖的分离纯化及组成结构分析

液体深层发酵所得的羊肚菌（Ｍｏｒｃｈｅｌｌａ ｅｓｓｕｌｅｎｔａ Ｌ．）营养液经超过滤法去掉小分子量组分,脱脂,脱色,脱蛋白质,减压低温浓缩,用超速离心法分离提纯,再用乙醇沉淀法反复多次将所要的多糖沉淀提纯,得到羊肚菌多糖（ＭＥＰ）。对其中分子量为１００００￣１０００００的多糖（ＭＥＰ－ＳＰ）,采用ＤＥＡＥ－纤维素离子交换柱,以不同浓度ＮａＣｌ为洗脱液,进行纯化分离,再经Ｓｅｐｈａｒｏｓｅ ＣＬ－     

当归水溶性多糖级分As-Ⅲa和As-Ⅲb的纯化鉴定与结构研究

当归热水抽提得到的粗多糖,经乙醇分级沉淀,ＤＥＡＥ纤维素分离和ＳｅｐｈａｄｅｘＧ１５０柱层析纯化,得到水洗脱级分Ａｓ—Ⅲａ和碱洗脱级分Ａｓ—Ⅲｂ两个多糖级分。经测定这两级分为均一组分。红外光谱呈现出典型的多糖吸收峰。气相色谱分析表明Ａｓ—Ⅲａ由葡萄糖组成,Ａｓ—Ⅲｂ由葡萄糖、甘露糖和阿拉伯糖组成。高碘酸氧化及Ｓｍｉｔｈ降解分析表明Ａｓ—Ⅲａ的单体通过α（１→３）糖苷键相连,Ａｓ—Ⅲｂ主要通过（１→４）和（１→６）糖苷键相连。     

水溶性甘草多糖的分离纯化、结构及生物活性研究进展

甘草是广泛使用的中药材之一.甘草中含有多种活性物质,甘草多糖作为主要的活性大分子,具有抗肿瘤、抗病毒、抗氧化、抗补体以及免疫调节等生物活性.本文对近年来水溶性甘草多糖的分离纯化、结构解析以及生物活性研究的最新进展进行综述.     

人参叶水溶性多糖——杂多糖P_N的纯化与结构初步确定

 从人参叶中提取的水溶性多糖经分离纯化得杂多糖P_N。P_N的分子量约为190万,单糖组成为阿拉伯糖、鼠李糖、木糖、半乳糖醛酸、半乳糖、葡萄糖及少量未知糖,单糖的摩尔比依次为8.1:0.8:1.0:1.6:12.5:4.1(未知糖除外)。经超离心分析,琼脂糖4B柱分析,玻璃纤维纸电泳和醋酸薄膜电泳鉴定等证明P_N为均一组份。经果胶酶降解,部分酸水解,高碘酸盐氧化,Smith降解,甲基化及其产物气相色谱(GLC)、气相色谱-质谱联用(GLC-MS)等结构分析表明P_N为多分支结构,分子的主链主要是由β-(1→3)连接的半乳糖组成,并在4—0和6—0上带有分支,平均每三个半乳糖有二个分支。     

白鲜皮多糖纯化、结构表征以及抗过敏研究

从白鲜皮(Cortex Dictamni)中分离纯化获得的白鲜皮多糖(Cortex Dictamni polysaccharides,CDPS),并经DE-AE柱分离得到两个多糖组分CDPS-1和CDPS-2,对其进行结构表征,同时考察CDPS的抗过敏活性.采用高效凝胶过滤色谱(GFC)、离子色谱(IC)、红外光谱(I...     

大马勃水溶性多糖的结构研究

采用热水浸提,乙醇沉淀的方法得到大马勃粗多糖,经Sevag法脱蛋白,DEAE—sepharose fast flow离子交换层析及sephacrylS-300HR凝胶过滤法得大马勃多糖（CGPI-1）。气相色谱研究表明,其单糖组成为：Gal,Glc,Man等四种单糖,其中Gal,Glc,Man,摩尔比为3．92：11．28：1．22。经部分酸水解,红外光谱及核磁共振光谱,高碘酸氧化,Smith降解等分析,CGPI-1的主链由Man和Glc构成,存在β型和α型两种糖苷键构型,支链或主链的末端残基由β-Gal（1→4）,β-Glc（1→6）,α—Glc（1→4）构成,其分子中存在酰胺结构。原子力显微镜观察发现CGPI-1呈分支的线性分子,在水溶液中容易互相缠绕形成强大的网络结构。     

苦豆子多糖的分离纯化及初步结构

本文研究了苦豆子多糖的分离纯化以及初步结构。采用水溶醇沉方法提取粗多糖,然后通过离子交换层析分离纯化,采用高效凝胶渗透色谱、气相色谱、紫外吸收以及红外吸收光谱对多糖组分进行分析研究。获得苦豆子多糖相对分子质量均一性组分SPN和SPA,测得其平均相对分子质量分别为1.217×106Da和4.412×105Da,二者均不含蛋白质,且都具有红外的多糖特征吸收峰,其单糖组成(质量比)分别为阿拉伯糖∶木糖∶甘露糖∶葡萄糖∶半乳糖=7.90∶11.24∶86.24∶18.81∶16.89、鼠李糖∶阿拉伯糖∶木糖∶甘露糖∶葡萄糖∶半乳糖=1.50∶5.80∶1.72∶20.10∶1.64∶27.20。本文为苦豆子多糖的进一步研究开发提供理论依据。     

Isolation, structural characterization and antioxidant activity of a new water-soluble polysaccharide from Acanthophyllum bracteatum roots

ABPS-1, a new water-soluble polysaccharide with molecular weight of 26 kDa and a specific optical rotation of +170° (c 1.0, H₂O), was extracted from the roots of Acanthophyllum bracteatum by warm water and further successively purified through DEAE-cellulose A52 and Sephadex G-100 columns. Monosaccharide analysis revealed that the ABPS-1 was composed of Glc, Gal and Ara with a relative molar ratio of 1.4:5.2:1.0. Its structural features were elucidated by a combination of FT-IR, methylation and GC-MS analysis, periodate oxidation and Smith degradation, partial acid hydrolysis and ¹³C and ¹H NMR spectroscopy. The data obtained indicate that ABPS-1 possessed a backbone of α-(1 → 6)-linked Gal with branches attached to O-2 by α-1 → linked Glc and at O-3 by α-1 → linked Gal and by α-(1 → 3)-linked Ara. The in vitro antioxidant activity showed that ABPS-1 possesses DPPH radical-scavenging activity in a concentration-dependent manner with an EC₅₀ value of 2.6 mg/ml.     

Hypoglycemic benefit and potential mechanism of a polysaccharide from Hericium erinaceus in streptozotoxin-induced diabetic rats

In this study, the hypoglycemic effect and possible mechanism of a polysaccharide, HEP-C, isolated from the fruit body of Hericium erinaceus were evaluated in streptozotoxin (STZ)-induced diabetic rats. Compared with the untreated STZ-induced diabetic rats, the supplements with HEP-C (150 and 300 mg/kg body weight [BW]) could significantly and dose-dependently relieve BW loss and organ injures, reduce fasting blood glucose, enhance glucose tolerance, alleviate hepatic function and serum lipid metabolism, elevate antioxidant enzyme activities, and suppress lipid peroxidation, which contributed to its potent hypoglycemic benefit. Liver histopathological observation revealed that HEP-C could effectively attenuate the deteriorated hepatic lesions in STZ-induced diabetic rats. HEP-C with potent hypoglycemic effect positively mediated glycogen synthesis by activating the phosphatidylinositol-3-kinase/protein kinase B signaling pathway. In summary, these results suggested that HEP-C, as a new dietary functional food or therapeutic agent, exhibited great potential for the prevention and treatment of diabetes mellitus and its complications.     

猴头菌和金针菇漆酶对不同染料的降解

利用漆酶(laccase)处理染料废水是近年来研究的热点.本研究以猴头菌Hericium erinaceus和金针菇Flammulina filiformis的发酵液为试验材料,通过硫酸铵沉淀、离子交换层析和超滤等方法,对发酵液中的漆酶进行了初步的分离纯化,然后分别研究了两种初提纯漆酶及其与小分子介体组成的漆酶介体系统...     

猴头菌UDP-葡萄糖-4-差向异构酶基因克隆表达及酶学性质

通过实验室前期对诱变菌株猴头菌321的多组学分析结果,获得了一个多糖合成过程中起关键作用的UDP-葡萄糖-4-差向异构酶基因（UDP-glucose-4-epimerase,UGE）,并在大肠杆菌E. coli BL21（DE3）中进行了异源表达。通过筛选最优的目的蛋白诱导表达条件后,通过镍柱亲和层析纯化后获得高纯度目的蛋白,并对目的蛋白进行了酶学性质的研究,明确了其生物学性质和动力学参数,为其开发利用提供理论参考。     

Isolation, purification and structural investigation of a water-soluble polysaccharide from Solanum lyratum Thunb

Polysaccharides, extracted from the herbs of Chinese natural Solanum lyratum Thunb (SLT), which is a traditional Chinese medicine with hot water and 0.04 M sodium hydroxide successively, were fractionated and purified by ion-exchange and gel-filtration chromatography. According to methylation, periodate oxidation, NMR spectroscopy, partial and graded acid and enzymic hydrolysis analysis, the results indicated the D-glucan to be linear and to contain both (1-->3)- and (1-->4)-linkages. The anomeric NMR and IR spectra measurements confirmed that the sugar residues were beta-glycosidically linked.     

猴头子实体锰型超氧物歧化酶的纯化及其性质鉴定

猴头子实体的超氧物歧化酶仅Ｍｎ－ＳＯＤ１种,有其资源和学术研究上的意义。其粗酶液经（ＮＨ４）２ＳＯ４盐析,ＤＥ５２和ＣＭ５２离子交换柱层析,纯化到电泳单斑点均一程度,其比活性为３０９６．５ｕ／ｍｇ,活性回收率为１４．８％。纯化的Ｍｎ－ＳＯＤ分子量为４７．０ＫＤ,亚基分子量为２３．３ＫＤ。金属元素分析表明每个亚基含１个Ｍｎ原子。该酶在紫外区有２个吸收峰,分别在２１８．４ｎｍ和２７６．０ｎｍ。该酶的理     

猴头多糖分离纯化的初步研究

目的：确定适合于猴头多糖分离纯化的方法。方法：以液体发酵生产的猴头菌丝为材料,提取猴头菌丝多糖进行分离纯化,以得到多糖纯品。结果：猴头菌丝粗多糖采用Sevag法除蛋白的次数应该控制在5-8次,而且Sevag法除蛋白所得的HMP,经DEAE-纤维素柱层析初步纯化,多糖主要分布在蒸馏水洗脱部分,命名为HMPⅠ,其含量为67.5%;HMPⅠ经Sephadex G-100凝胶柱层析纯化,得到两个组分：HMPⅠa、HMPⅠb;HMPⅠa为多糖主要组分,含量为71.8%;HMPⅠa经纯度鉴定为多糖纯品。结论：DEAE-纤维素柱层析结合Sephadex G-100凝胶柱层析的纯化方法,可以获得猴头多糖纯品。     

Isolation and structural characterization of a neutral polysaccharide from the stems of Dendrobium densiflorum

A novel neutral heteropolysaccharide (DDP-1-D) was purified from hot water extracts of dried stem of Dendrobium densiflorum by DEAE-52 and Sephacryl S-200 High-Resolution Chromatography. The heteropolysaccharide had an average molecular weight about 9440 Da. It was composed mainly of glucose and mannose in the ratio of 3.01:1. Structural features of DDP-1-D were elucidated by a combination of chemical and instrumental techniques, including FT-IR, GC-MS, periodate oxidation-Smith degradation, (1)H and (13)C NMR spectroscopies (including COSY, TOCSY, HSQC, and HMBC spectra). The results indicated that DDP-1-D is a mannoglucan and has a backbone consisting of (1→4)-linked α-D-Glcp, (1→6)-linked α-D-Glcp, (1→2)-linked α-D-Manp and (1→4)-linked β-Manp. This is the first study to provide clear evidence for the structure of the polysaccharide in D. densiflorum.     

Purification and partial characterization of a novel hemagglutinating glycoprotein from the cultured mycelia of Hericium erinaceus

HEG-5, a novel glycoprotein with hemagglutinating activity, was firstly isolated and purified from the cultured mycelia of Hericium erinaceus CZ-2. SDS–PAGE, Native-PAGE and MALDI-TOF-MS proved that HEG-5 was a single band with the molecular weight of approximately 14.4 kDa. HEG-5 had the protein: polysaccharide ratio of approximately 10:1 (%/%) and contained d-glucose, l-rhamnose, d-galactose and d-mannose with a molar ratio of 1.00:1.09:2.45:7.14 in polysaccharide fraction. HEG-5 was an acidic glycoprotein with a PI value of 6.3 and the higher content of acidic amino acids (Asp, 12.42 ± 0.25% and Glu, 12.24 ± 0.26%) in protein fraction. FT-IR and NMR spectra revealed that HEG-5 contained the protein and carbohydrate portions with (1→4)-linked β-galactose residues and β-linked glucose residues. Circular dichroism (CD) demonstrated that HEG-5 was a β-sheet predominant glycoprotein. Hemagglutination assay proved it was a thermo-unstable glycoprotein. The HEG-5 structural novelty was finally presented by protein sequencing and modeling by using MALDI-TOF-MS, NCBI blast search and online SWISS-MODEL Workspace service.     

Purification and structural characterization of a new water-soluble neutral polysaccharide GLP-F1-1 from Ganoderma lucidum

A water-soluble neutral polysaccharide (GLP-F1-1) was isolated from the fruiting bodies of Ganoderma lucidum by DEAE Sepharose Fast Flow and Sephacryl S-500 High Resolution Chromatography. The neutral polysaccharide had an average molecular weight (Mw) of approximately 2.5×10(6) kDa. GC analysis showed that this polysaccharide was mainly composed of glucose and galactose in the molar ratio of 34:1. 1H and 13C NMR spectroscopy in combination with GC-MS technique indicated that the new polysaccharide had a backbone chain of 1,4-disubstituted-β-glucoseopyranose and 1,4,6-trisubstituted-β-glucoseopyranosyl, while the branched chains were mainly composed of 1,6-disubstituted-β-glucopyranosyl and 1,4-disubstituted-β-galactoseopyranosyl residues.