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1.
条斑紫菜(Porphyra yezoensis)的热水提取物,经DE-23和Sephadex G-200柱层析纯化,得到一种含微量硫酸基的琼胶精品(PY1)。此多糖经Sepharose 6B柱层析鉴定为单一组分,分子量为220 kD。紫外光谱显示它不含蛋白质、多肽及核酸。红外光谱揭示它含3,6_内醚_半乳糖的特征吸收以及微量的硫酸基。该多糖的水解产物经气相色谱分析,主要由半乳糖及其衍生物组成,有少量岩藻糖存在。通过高磺酸氧化和Smith除解以及13C-NMR谱的分析,该多糖主要由琼胶二糖结构单元和它的生物前体组成,以琼胶二糖为主。  相似文献   

2.
浒苔多糖的分离、纯化和分析   总被引:1,自引:0,他引:1  
浒苔(Enteromorphaprolifera)经热水提取,Sevage法除去蛋白质,用乙醇沉淀,SephadexG-100柱层析,得浒苔多糖(简称EP)精制品。经SephadexG-200柱层析鉴定为单一对称性洗脱峰。红外光谱分析具有多糖特征吸收峰,紫外光谱分析未见有核酸和蛋白质的特征吸收峰。总糖含量为88.8%,其中糖醛酸含量为33.6%。单糖组成为L-阿拉伯糖、L-岩藻糖、D-甘露糖、D-半乳糖及D-葡萄糖,平均分子量为25000。  相似文献   

3.
浒苔多糖的分离,纯化和分析   总被引:15,自引:0,他引:15  
浒苔经热水提取,Sevage法除去蛋白质,用乙醇沉淀,Sephadex-G-100柱层析,得浒苔多糖(简称EP)精制品。经SephadexG-200柱导析鉴定为单一对称性洗脱峰。红外光谱分析具有多糖特征吸收峰,紫外光谱分析表未见有核酸和蛋白质的特征及吸上峰,总糖含量为88.8%,基中糖酸酸含量为33.6%。单糖组成为L-阿拉伯糖,L-岩藻糖,D-甘露糖,D-半乳糖及D-葡萄糖。平均分子量为2500  相似文献   

4.
液体发酵得到的红栓菌(Pycnoporuscinnabarius)菌丝经沸水抽提,胰匀浆处理和Sevage法除蛋白,DEAESephadexA-25与SephadexG-150柱层析,乙醇沉淀得到二种多糖组分(简称PC_1、PC_2)。经聚丙烯酰胺电泳,冻融分析鉴定为均一体。PC_1和PC_2的,总糖含量分别为81.5%和77.3%,分子量分别为23000和13000.两者的红处光谱具多糖特征吸收峰,在紫外区无明显吸收现象,基本不含氮。PC_1单糖组成为D-葡萄糖、D-半乳糖及D-甘露糖,PC_2为D-半乳糖、D-甘露糖。  相似文献   

5.
云南溪菜多糖的分离纯化及其性质研究   总被引:8,自引:0,他引:8  
从云南溪菜中提取的水溶性胞外粗相糖(EPS),用Sevag法脱蛋白后,经DEAE-纤维素柱层析和葡萄糖凝胶G-200柱层析可得到纯品。溪菜多糖复合物经碘试验证明含有少量淀粉,Bacl2试验检测出含有SO4^2-,琼脂糖凝胶电泳显示含有糖类及蛋白南,且其组成不均一。用Lowry法测得蛋白质含量为13.54%,苯酚-硫酸法测得糖含量为78%,经薄层层析和纸层析鉴定,溪菜多糖复合物的酸水解物中含有大量D  相似文献   

6.
尖吻蝮蛇酸性磷脂酶A2的纯化和初步晶体学   总被引:5,自引:0,他引:5  
为进一步揭示影响血小板聚集活性的结构基础,纯化了江西尖吻蝮蛇(Agkistradun acutus)酸性磷脂酶A2。江西省吻蝮蛇蛇毒经CM-Sepharose离子交换柱层析,两步DEAE-Sepharose离子交换柱层析以及Mono)GFPL离子交换柱层析,得到了SDS聚丙烯酰胺凝胶电泳和等和集电泳均一的磷脂酶A2(PLA2),其分子量为16.5KD,等电点为4.3,经测定,该酶具有抑制ADP诱地  相似文献   

7.
以「^35S」-Na2SO4为示踪物,观察人正常主动脉中的硫酸乙酰肝素蛋白聚糖(HSPG)对培养的第一代人脐静脉内皮细胞(hUVEC)合成蛋白聚糖(PG)的影响,用解聚提取法及离子交换柱层析分离人主动脉HSPG,^35S-PGs的混合物用离子交换及凝胶过滤柱层析法分离^35S-HSPG,^35S-硫酸软骨素-硫酸皮肤素PG(^35S-CSDSPG)及^35S-硫酸皮肤素PG(^35S-DSPG),  相似文献   

8.
两种黄精多糖衍生物的制备及其抗病毒活性比较研究   总被引:13,自引:0,他引:13  
从传统中药黄精属的两个种中分别提取到两种小分子粗多糖(PD,PP),用吡啶-氯磺酸法分别制取到这两种粗多糖的硫酸酯(PDS,PPS)。产物经红外、紫外鉴定,证实多糖羟基上连上了硫酸基。用离子交换,凝胶色谱提纯得到这两种粗多糖的纯化产物(RPD,RPP)。体外实验研究了这6种物质对非洲绿猴肾细胞(Vero)的细胞毒性和抗单纯疱疹病毒2型(HSV-Ⅱ)的活性。结果表明,粗多糖PP的细胞毒性是由其中的杂  相似文献   

9.
以对硝基苯糖苷基为底物,测定了慈菇的12种糖苷酶,其中α-甘露糖苷酶、α-和β-半乳糖苷酶活力较高;经硫酸铵分级沉淀,SephadexG-150分子筛层析,ConASepharose4B亲和层析,DEAE-SepharoseCL-6B离子交换层析,从慈菇抽提液纯化了α-半乳糖苷酶。纯化酶的比活提高1072倍,活力回收15.6%,在圆盘聚丙烯酰胺凝胶电泳和SDS-PAGE上均显示1条蛋白质带,在α-半乳糖苷酶浓度为150mU/ml的溶液中测不到其他糖苷酶的活力。慈菇α-半乳糖苷酶的分子量用SephadexG-100凝胶过滤柱测定或在SDS-PAGE上测定均为60kD,酶反应的最适pH在5.8附近,最适温度为60℃。该酶分解对硝基苯基-α-半乳糖苷的K_m值为3.7×10 ̄(-4)mol/L,V_m值为2.1×10 ̄(-4)mol/L。银离子、汞离子显著抑制酶活力,D-半乳糖和密二糖均竞争性地抑制该酶水解对硝基苯基α-D-半乳糖苷的活力,根据Dixon作图求得其K_i值分别为0.92×10 ̄(-3)mol/L和1.98×10 ̄(-3)mol/L。2-脱氧-D-半乳糖和L-岩藻糖为酶活力的非竞争性抑制剂。化学修饰  相似文献   

10.
当归多糖的分离化及其部分性质的研究   总被引:7,自引:0,他引:7  
中药当归经热水提取、乙醇分级沉淀、乙醇分级沉淀、DEAE纤维素和SepadexG-150柱层析,得As-Ⅲb两个多糖级份。As-Ⅲa和AsⅢb经醋纤维薄膜电泳、玻璃纤维纸电泳和凝胶柱层析分析,证明都是均二的,经琼脂糖凝胶柱层析测得As-Ⅲa和AsⅢb的分子量分别为85KD和49KD左右。组成分析表明As-Ⅲa由葡萄糖组成,As-Ⅲb由葡萄糖、甘露糖和阿拉伯糖组成,三者比为10:10:4。  相似文献   

11.
一种紫菜多糖的制备及对淋巴细胞生长的影响   总被引:6,自引:0,他引:6  
用DEAE-纤维素和SephadexG-200柱层析法分离纯化条斑紫菜的热水提取物,从中得到多糖PY2,并测出其分子量为2.0%10^4。用紫外和红外光谱对PY2的性质进行了鉴定。进一步测定了PY2对体外培养的小鼠骨髓细胞及淋巴细胞生长的影响。结果表明,PY2是一种少见的紫菜多糖,它不含有大多数紫菜侈糖具有的3,6-内醚-兰乳糖和硫酸基,它对小鼠脾脏淋巴细胞、胸腺淋巴细胞以及混合淋巴细胞的增殖有一定的抑制作用,而对骨髓细胞的增殖没有明显的影响。  相似文献   

12.
从淫羊藿中提取多糖并鉴定其初步结构和单糖组成.采用超声-水提醇沉法提取粗多糖、Sevag法去蛋白、DEAE-52纤维素及Sephadex G-100柱层析法纯化得到淫羊藿多糖EPSⅠ-1和EPSⅡ-1.应用紫外光谱法和红外光谱法对其结构做初步分析.采用高效液相色谱法(HPLC)测定其单糖组成及摩尔比.均一的EPSⅠ-1和EPSⅡ-1多糖在紫外和红外中具有多糖的特征吸收峰,组成中含有吡喃环结构;EPSⅠ-1的单糖组成为鼠李糖和葡萄糖,摩尔比为1:1.13;EPSⅡ-1的单糖组成为果糖、葡萄糖和一个不确定的糖,摩尔比为1:1.91.有效地分离纯化了淫羊藿多糖,这为淫羊藿多糖的广泛应用奠定了实验基础.  相似文献   

13.
Purification and characterization of murine protoporphyrinogen oxidase   总被引:8,自引:0,他引:8  
H A Dailey  S W Karr 《Biochemistry》1987,26(10):2697-2701
The penultimate enzyme of the heme biosynthetic pathway, protoporphyrinogen oxidase (EC 1.3.3.4), has been purified to apparent homogeneity from mouse liver mitochondria. The purification involves solubilization from mitochondrial membranes with sodium cholate followed by ammonium sulfate fractionation and gel filtration on a Sepharose CL-6B column. The eluate is adjusted to 0.67 M (NH4)2SO4 and loaded onto a phenyl-Sepharose column. After salt washes, the enzyme is eluted with 0.5% sodium cholate and 0.5% Brij 35. The final step is high-pressure ion-exchange chromatography on a DEAE-5PW column. The purified protein has a molecular weight of approximately 65,000 by gel filtration chromatography on Sepharose CL-6B in the presence of 0.5% sodium cholate. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows a single band corresponding to a molecular weight of 65,000. The absorption spectrum of the purified enzyme shows no evidence of a chromophoric cofactor. Purified protoporphyrinogen oxidase has a Km for protoporphyrinogen IX of 5.6 microM with a Vmax of 2300 nmol mg-1 h-1. It utilizes meso- and hematoporphyrinogen at about 10% the level of protoporphyrinogen. The pH optimum is broad with a maximum at 7.1. There is no stimulation or inhibition by any tested divalent cations, and sulfhydryl reagents have no inhibitory effect on the purified enzyme.  相似文献   

14.
一种新的蘑菇凝集素的生化特征和氨基酸分析   总被引:4,自引:0,他引:4  
用 (NH4) 2 SO4分级沉淀、离子交换和分子筛等方法 ,从食用蘑菇中分离纯化出一种凝集素 ,经SDS -PAGE测定其亚基的相对分子量为 15 .8kDa,质谱分析法测定其分子量为 32kDa ,说明该凝集素由两个亚基组成。氨基酸分析发现 ,该凝集素不含半胱氨酸。热稳定试验和耐酸碱试验显示 ,该凝集素活性具很高稳定性  相似文献   

15.
A large number of polysaccharides are present in boiling-water extraction of Dioscorea nipponica Makino. A DEAE-Sepharose CL-6B column chromatography was used to isolate the major polysaccharides from D. nipponica Makino. The largest amount of fraction of polysaccharide was subjected to further purification by gel-filtration on Sephadex G-100. The purified fraction was a neutral polysaccharide and a single peak in HPLC with Sugar KS-804 column, with a molecular weight of 38,000, and comprised mainly of glucose and fructose (45:1). Analysis by Periodate oxidation–Smith degradation indicated that there were 5.9%(1→)-glycosidic linkages, 4.94% (1 → 2)-glycosidic linkages, 61.16% (1 → 4)-glycosidic linkages, and 28% (1 → 3)-glycosidic linkages. On the basis of superoxide radical assay, hydroxyl radical assay, and self-oxidation of 1,2,3-phentriol assay, its antioxidant activity was investigated. This purified fraction of polysaccharide exhibited equivalent inhibiting power for self-oxidation of 1,2,3-phentriol to Vc, a little higher scavenging activity of superoxide radical and hydroxyl radical than Vc, and should be explored as a novel potential antioxidant.  相似文献   

16.
A protein recognizing apolipoproteins AI, AII and AIV was purified from cultured mouse adipose cells of the Ob17MT18 clonal line. Apolipoprotein A binding sites were solubilized in the presence of proteinase inhibitors using the non-denaturating detergent CHAPS. Chromatography of the soluble extract on DEAE-Trisacryl was followed by immunoaffinity chromatography of the complex apolipoprotein AI-binding proteins on anti-(apolipoprotein AI) coupled to Sepharose 4B and then by h.p.l.c. on an RP-Select B column. A 1400-fold purification over the starting crude homogenate was achieved. The purified material contained two proteins that were both able to bind apolipoproteins AI, AII and AIV, but not low-density lipoprotein. Glycopeptidase F treatment showed the existence of a single protein bearing either N-linked high-mannose or complex oligosaccharide chains. The purified material showed an apparent molecular mass of 80 +/- 9 kDa by h.p.l.c. on a TSKG 3000 SW column. Rabbit polyclonal antibodies directed against the purified material revealed two protein bands of 80 and 92 kDa after SDS/PAGE under reducing conditions and immunoblotting. These bands were undetectable in growing Ob17PY cells previously shown not to bind the various apolipoproteins A and not to undergo cholesterol efflux, whereas they were conspicuous in growth-arrested Ob17PY cells which have recovered these properties.  相似文献   

17.
1. NADH-cytochrome b5 reductase was purified from sheep lung microsomes in the presence of non-ionic and ionic detergents, Emulgen 913 and cholate, respectively. 2. The purification procedure involved the ion-exchange chromatography of the detergent solubilized microsomes on DEAE-cellulose. 3. Further purification and concentration of lung reductase was carried out with a second DEAE-cellulose column followed by the affinity column chromatography of partially purified reductase on 5'-ADP-agarose column. 4. The specific activity of sheep lung reductase was 638 mumol ferricyanide reduced/min/mg protein and the yield was 6% of the initial activity in microsomes. 5. The SDS-polyacrylamide gel electrophoresis of the purified lung reductase showed one protein band having the monomer mol. wt of 34,500 +/- 1500. In the presence of 0.4% deoxycholate, it existed as an active dimer having a mol. wt of 68,500. 6. Trypsin treated lung reductase showed two extra protein bands of mol. wts of 28,000 and 25,000 on 10% SDS-polyacrylamide gels. 7. The purified enzyme was found to contain FAD as prosthetic group and the absorption spectrum of lung reductase showed two peaks at 390 and 461 nm which were typical for flavoproteins and a shoulder at 490 nm. 8. The maximal activity of lung reductase was observed between pH 6.5-8.0 and at pH 6.8, when ferricyanide and partially purified sheep lung cytochrome b5 was used as electron acceptors, respectively.  相似文献   

18.
羊栖菜褐藻糖胶抗凝血活性的研究   总被引:9,自引:2,他引:7  
本文研究了羊栖菜褐藻糖胶的化学组成和抗凝血活性之间的关系。采用热水提取得羊栖菜粗多糖,CaCl2纯化得褐藻糖胶,DEAE Sepharose CL-6B柱层析与Sepharose CL-6B柱层析对褐藻糖胶进行分级,得到F1、F2、F31、F32和F33五个级分,均为岩藻糖、半乳糖和甘露糖等糖基组成的杂多糖,并含有硫酸酯和糖醛酸以及少量的蛋白质,相对分子质量范围2.5万~95万。采用活化部分凝血活酶时间(APTT)和凝血酶时间(TT)检测了这5个级分的抗凝血活性,结果显示,羊栖菜褐藻糖胶能显著延长APTT的凝血时间,而对TT的影响不明显。F1、F31和F32对APTT的影响比较显著,而F2、F33和羊栖菜粗多糖的影响较小。研究表明,羊栖菜褐藻糖胶主要是通过抑制内源凝血途径而达到抗凝血的效果,其抗凝血活性与褐藻糖胶的硫酸基含量成正相关,而与相对分子质量和糖醛酸含量无关。  相似文献   

19.
Clostridium botulinum type E toxin was purified in three chromatography steps. Toxin extracted from cells was concentrated by precipitation and dissolving in a small volume of citrate buffer. When the extract was chromatographed on DEAE-Sephadex without RNase or protamine treatment, the first protein peak had most of the toxin but little nucleic acid. When the toxic pool was applied to a carboxymethyl Sepharose column, toxin was recovered in the first protein peak in its bimolecular complex form. The final chromatography step at 4 degrees C on a DEAE-Sephacel column at a slightly alkaline pH purified the toxin (Mr, 145,000) by separating the nontoxic protein from the complex. At least 1.5 mg of pure toxin was obtained from each liter of culture, and the toxicity was 6 X 10(7) 50% lethal doses per mg of protein. These values are significantly higher than those previously reported.  相似文献   

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