摘要
本文通过对灰树花β-葡聚糖的提取、测定、纯化和结构的研究,建立了荧光法测定1,3-β-葡聚糖的方法,并探讨了其测定影响因素;确定了灰树花β-葡聚糖高效提取的条件;分析了一种灰树花β-葡聚糖的结构。
1.研究表明可得然1,3-β-葡聚糖、酵母1,3/1,6-β-葡聚糖及灰树花中1,3-链葡聚糖与苯胺蓝的结合具有特异性;当以酵母1,3/1,6-β-葡聚糖为标准物时蛋白浓度在葡聚糖浓度的一半以下,蛋白对测定影响不大;当以可得然凝胶为标准物测量时,蛋白对测定影响不大,淀粉,纤维素和麦芽糊精添加会导致两种测定的荧光强度下降。
2.食用菌1,3-β-葡聚糖荧光法测定与鲎试剂法测定比较,结果表明荧光法简单可行。
3.通过响应面优化,得到灰树花多糖提取最佳条件为:提取功率137.6W;超声时间为16.9 min;酶解温度40℃,时间45 min,pH 6.0,加酶量1.5%;95℃提取两次,在此条件下,10 g灰树花多糖提取得理论值达到12.75%;灰树花β-葡聚糖提取最佳条件为:提取频率为148.3 W;时间15.6 min;温度为95℃,在此条件下,10 g灰树花多糖提取得理论值达到3.60%。
4.对热水提取粗β-葡聚糖进行离子交换层析、凝胶柱层析,纯化得到一种均一多糖GFWD,通过对其进行单糖组分分析,红外光谱分析及核磁共振分析,确定此多糖是一种1,3-β-葡聚糖。
本研究创新点在于:
1.建立了荧光法测定1,3-β-葡聚糖的方法,并研究了其影响因素。
2.以响应面法优化了灰树花β-葡聚糖的超声-酶法提取工艺。
3.测定了灰树花热水提、冷碱提取和热碱提取粗多糖中β-葡聚糖和1,3-β-葡聚糖的含量。
4.通过气相色谱、红外光谱和核磁共振解析了一种热水提取β-葡聚糖的结构。
Systematic studies on the extraction,quantificational,purification and structural characterization of Grifola frondosa polysaccharides and itsβ-glucan were carried out in the presented dissertation.Some new methods and points were developed on the basis of thesis of these studies.The results and creative points are described as follows:
1.The result showed curdlan,1,3/1,6-β-glucan of yeast and Grifola frondosa 1,3-β-glucan could bind to Aniline Blue specifically,when the content of the protein was the half of the content of 1,3/1,6-β-glucan,there was no effect on quantificational with the standard 1,3/1,6-β-glucan of yeast.The protein no effect on quantificational with the curdlan standard.Starch,the dextrin,glucose and cellulose no effect.
2.To contrast to detecting method about 1,3-β-glucan about Limulus factor G method and fluorescence method,the result show fluorescence method was simple and feasible.
3.By response surface methodology,the optimum polysaccharides conditions of ultrasonic and enzyme assisted extraction was that ultrasonic power was 137.6 W,ultrasonic time was 16.9 min;the optimumβ-glucan conditions was ultrasonic power was 148.3 W. ultrasonic time was 15.6 min.Hydrolyzed temperature was 40℃. hydrolyzed time was 45 min,pH was 6.0 and the quantity of papain was 1.5%of raw material,then extraction was twice under 95℃.By the conditions,the extraction rate of polysaccharides andβ-glucan were 12.75%and 3.60%.
4.Separated the crudeβ-glucan by anion exchange and gel chromatographic,then to analyze the homogeneous sugar and structure of aβ-glucan from Grifola frondosa by hot water through High Performance Liquid Chromatography(HPLC),Gas Chromatography (GC),Infrared spectroscopy(IR)and Nuclear Magnetic Resonance (NMR).
The innovations that originally studies are embodied in the following several points:
1.A simple detecting method about 1,3-β-glucan by fluorescence method established according to some scientific reports.
2.To found the optimum conditions of ultrasonic and enzyme assisted extraction by hot water.
3.Determinated of Grifola frondosa 1,3-β-glucan by hot water,cold alkali and hot alkali.
4.To analyze structure of aβ-glucan from Grifola frondosa by hot water through GC,IR,NMR.
引文
[1]D.A.Rees Outline Studies in Biology Polysaccharide Shapes[M].1982.
[2]阿历索保罗等著,姚一建等译.菌物学概论(第四版)[M].中国农业出版社,2002.
[3]Qun Dong,Lian-meng Jia,Ji-nian Fang.A β- glucan isolated from the fruiting bodies of Hericium erinaceus and its aqueous conformation[J].Carbohydrate Research,2006,341:791-795.
[4]M anzi.P amel,P izzoferrato,L aura.Beta-glucans in edible mushrooms[J].Food chemistry,2000,68:315-318.
[5]Elaine R,Carbonero,Ana Helena P.Gracher,Fhemanda R,Smiderle,Fabio R,Rosado,Guilherme L.Sassaki,Philip A.J.Gorin,Marcello Iacommi.A β-glucan from the fruit bodies of edible mushrooms Pleurotus eryngii and Pleurotus ostreatoroseus[J].Carbohydrate Polymers,2006,252-257.
[6]陈倩,穆晓峰.香菇多糖生物活性研究进展[J].甘肃教育学院学报(自然科学版).2003,17(4):27-30
[7]H.Nanba,A.Hamaguchi,H.Kuroda.The chemical structure of an antitumor polysaccharide in fruit bodies of Grifola frondosa(Maitake)[J].Chem.Pharm.Bull,1987,35(3):1162-1168.
[8]H.Kawagishi,Ryuichi Inagaki,Teturo Kanao,Takashi Mizuno.Fractionnation and activity of the water-insoluble residue of Agaricus blazei friuiting bodies[J].Carbodydrate Research.1989,186:267-273.
[9]H.Kawagishi,T.Kanao,R.Inagaki,T.Mizuno,K.Shimura,H.Ito,T.Hagiwara.T.Nakamura.Formolysis of a potent antitumor(1→6)-β-D-glucan-protein complex from Agaricus blazei fruiting bodies and antimmor activity of the resulting products[J].Carbohydrate Polymers,1990,12(3):393-403
[10]Falch,Berit H,Espevi k,Terje,Ryan,et al.The cytokine stimulating activity of β-(1-3)-D-glucans is dependent on the triple helix conformation[J].Carbohydrate Research.2000.329:587-596
[11]Lina Zhang,Xuelian Li,Xiaojuan Xua,Fanbo Zeng.Correlation between antitumor activity.molecular weight and conformation of lentinan[J].Carbohydrate Research,2005,340:1515-1521.
[12]Shih-Houng Young,Robert,R.Jacobs.Sodium hydroxide-induced conformational change in schizophyllan detected by the fluorescence dye,aniline blue[J].Carbohydrate Research,1998,310:91-99.
[13]Unursaikhan Surenjav,Lina Zhang,Xiaojuan Xu,Xufeng Zhang,Fanbo Zeng.Effects of molecular structure on anti-tumor activities of(1,3)-β-D-glucans from different Lentinus Edodes[J].Carbohydrate Polymers,2006,63(1):97-104.
[14]Yifeng Wang,Lina Zhang,Yunqiao Li,Xiaohua Houb,Fanbo Zeng.Molecular mass and antitumor activities of sulfated derivatives of β-glucan from Poria cocos mycelia[J].International Journal of Biological Macromolecules,2004,34:231-236.
[15]Yongzhen Tao,Lina Zhang,Peter C.K.Cheung.Physicochemical properties and anti-tumor activities of water-soluble native and sulfated hyperbranched mushroom polysaccharides[J].Carbohydrate Research,2006:341:2261-2269.
[16]Borchers AT,Stem JS,Hackman RM,Keen CL,Gershwin ME.Mushroom,tumors.immunity[J].Proc Soc Exp Biol Med,1999,221(4):281-293.
[17]Guterrez ZR,Mantovani MS,Eira AF,Ribeiro LR,Jordao BQ.Variation of the antimutagenicity effects of water extracts of Agaricus blazei Murrill in vitro[J].Toxicol.In Vitro.2004,18:301-309.
[18]Yanfei Peng,Lina Zhang,Fanbo Zeng and John F.Kennedy.Structure and antitumor activities of the water-soluble polysaccharides from Ganoderma tsugae mycelium[J].Carbohydr.Polym,2005,59(3),385-392.
[19]R Zheng,S Jie,D Hanchuan,W Moucheng.Characterization and immunomodulating activities of polysaccharide from Lentinusedodes[J].Int Immunopharmacol,2005.5:811-820.
[20]Ukawa Y.Ito H,Hisamatsu M.Antitumor Effects of(1-3)-β-D-Glucan and(1-6)-β-D-Glucan Purified from Newly Cultivated Mushroom,Hatakeshimeji(Lyophyllum decmtes Sing.)[J].Journal of bi osci ence and bioencineering,2000,90(1):98-104.
[21]Kodama N.Murata Y,Asakawa A,Inui A,Hayashi M.Maitake D-Fraction enhances anti-tumor effects and reduces immunosuppression by mitomycin-C in rumor-bearing mice [J].Nutrition,2005,21:624-629.
[22]Otagiri K,Ohkuma T,Ikekawa T,Tanaka S.Intensification of antitumor-immunity by protein-bound polysaccharide EA6,derived from Flammulina velutipes(Curt.ex Fr)Sing.combined with murine leukemia L1210 vaccine in animal experiments[J].J Pharmacobiodvn.1983,6:96-104.
[23]TA Ajith,KK Janardhanan.Cytotoxic and antitumor activities of a polypore macrofungus,Phellinus rimosus(Berk)Pilat[J].J Ethnopharmacol,2003,84:157-162.
[24]Chang-Qing Gu,Juan-Wen Li,Fu-Huan Chao.Inhibition of hepatitis B virus by D-fraction from Grifola frondosa:Synergistic effect of combination with interferon-ct in HepG2 2.2.15[J].Antiviral Research,2006,72:162-165.
[25]Goksel Sener,Hale Z.Toklu,Sule Cetinel.β-Glucan protects against chronic nicotine-induced oxidative damage in rat kidney and bladder[J].Environmental Toxicology and Pharmacology,2007,23:25-32.
[26]Sener G,Toklu H,Ercan F,Erkanli G.Protective effect of β-glucan against oxidative organ injury in a rat model of sepsis[J].International Immunopharmacology,2005,5:1387-1396.
[27]Sener G,Sert G,Ozer Sehirli A,Arbak S,Uslu B.Pressure ulcer-induced oxidative organ injury is ameliorated by β-glucan treatment in rats[J].International Immunopharmacology.2006,6:724-732.
[28]Susanne Alban,Walter Jeske,Dieter Welzel,Gerhard Franz,Jawed Fareed.anticoagulant and antithrombotic actions of a semisynthetic β-1,3-glucan sulfate[J].Thrombosis Research.1995,78(3):201-210.
[29]Juliana Curi Martinichen-Herrero,Elaine Rosechrer Carbonero,Philip Albert James Oorin.Marcello Iacomini.Anticoagulant and antithrombotic activity of a sulfate obtained from a glucan component of the lichen Parmotrema mantiqueirense Hale[J].Carbohydrate Polymers.2005,60:7-13.
[30]谢红旗,周春山,杜邵龙.酶法提取香菇多糖新工艺研究.食用菌.2006(4):59-61
[31]P Valachovi,A Pechova,T J Mason.Towards the industrial production of medicinal tincture by ultrasound assistedextraction[J].Ultrason.Sonochem.2001(8):111-117.
[32]Z Hromadkova,A Ebringerova,P Valachovi.Ultrasound assisted extraction of water- soluble polysaccharides from theroots of valerian[J].Ultrason Sonochern,2002(9):37-44.
[33]黄永春,马月飞,谢清若等.超声波辅助提取茶多糖及其分子量变化的研究.食品科学。2007,28(7):170-173
[34]廖周坤,姜继祖,王化远等.超临界CO2萃取藏药雪灵芝中总皂苷及多糖的研究[J].中草药,1998,29(9):601.
[35]Mark Mayell.Maitake Extracts and Their therapeutic Potential-A Review.Alternative Medicine Review.2001,6(1)
[36]Jones K.Maitake:a potent medicinal food.Alt Comp Ther.1998,4:420-429
[37]Takama F,Ninomiya S,Yoda R,et al.Parenchyma cells,chemical components of maitake mushroom(Grifola frondosa S.F.Gray)cultured artificially,and their changes by storage and boiling.Mushroom Sci.1981,11:767-779
[38]Chang R.Functional properties of edible mushroom.Nutr Rev.1996,54:s91-s933
[39]Nanba H,Hamaguchi A,Kuroda.The chemical structure of an antitumor polysaccharide in fruit bodies of Grifola frondosa(maitake).Chem Pharrn Bull.1987,35:1162-1168.
[40]Hisatora K.et al.Glucan having beta-1,6 bond-containing main chain,obtained from maitaike and antineoplastic agent comprising same.Japanese Patent JP59210901.1984.
[41]Adachi Y,Ohno N,Ohsawa M,et al.Change of biological activies of 1,3-beta-D-glucan from Grifola frondosa upon molecular weight reduction by heat treatment.Chem Pharm Bull.1990.38:477-481.
[42]Okazaki M,Adachi Y,Ohno N,Yadome T.Structure-activity relationship of 1,3-beta-D-glucans in the induction of cytokine production from macrophages,in vitro.Biol Pharm Bull.1995,18:1320-1327
[43]Nanba H,Kubo K.Antitumor substance extracted from Grifola.U.S.Patent 5,854,404,1998
[44]Adachi Y,Okazaki M,Ohno N,Yadome T.Enhancement of cytokine production by macrophages stimulated with 1,3-beta-D-glucan,grifolan(GRN),islated from Grifola frondisa.BiolPharm Bull.1994,17:1554-1560
[45]Ohno N,Asada N,Adachi Y,Yadomae T.Enhancement of LPS TNF-alpha(tumor nedrosis factor-alpha)production by 1,3-beta-D-glucans in mice.Biol-Pharm bull.1995,18:126-135
[46]Kurashige S,Akuzawa Y,Endo F.Effects of Lentinus edodes,Grifola frondosa and Pleurotus administration on cancer outbreak,and activities of macrophage and lymphocytes in mice treated with a carainogen,N-butyl-N- butanolnitrosoamme.Immunopharmocol Immunopharmacol immunotoxicol.1997,19:175-183.
[47]Miutra N,Ohno N,Aketagawa J,et al.Blood clearance of 1,3-beta-D-glucan in MRL mice.FEMS Immunol Med Microbiol.1996,13:51-57
[48]Ohno N,Suzuki 1,SatoK et al.Purification and structural characterization of an antirumor β-1,3-Glucan isolated from hot water extract of the fruit body of cultured Grifola frondosa.Chemical and Pharmaceutical Bulletin,1985,33(10):4522-452
[49]Ohno H,Adachi Y,SuzukiI et al.Characterization of the antitumor glucan obtained from liquid-cultured Grifola frondosa.Chemicala nd Pharmaceutical Bulletin.1986,34(4):1709-1715
[50]Ohno N,Hayashi M,SuzukiI et al.Efect of glucans on the antitumor activity of Grifolan.Chemical and Pharmaceutical Bulletin.1986,34(5):2149-2151
[51]Nanba H.Antitumor activity of oral administered "D-Fraction" from Maitake mushroom (Grifola frondosa).J Nat Med.1993,1(4):10-15
[52]Hishida I,Nanba H,Kuroda H.Antitumor activity exhibited by orally administered extracted from fruit body of Orifola frondosa.Chemical and Pharmaceutical Bulletin.1988.36(5):1819-1827
[53]Naomi Miyake,Yuki Masuda,Yukihito Murata et al.Low molecular weight maitake MD-Fraction(Klasma-MD)hydrolyzed with endo-b-1,6-glucanase of Trichodenna harzianum induces antitumor activities.Mycoscience.2006,47:78-83
[54]Noriko Kodama,Ph.D.a,Yukihito Murata et al.Maitake D-Fraction enhances antitumor effects and reduces immunosuppression by mitomycin-C in tumor-bearing mice.Nutrition.2005,21:624-629
[55]张劲松,樊华,李玉祥等.几种药用真菌粗提物对多种人体肿瘤细胞株增殖的抑制作用.天然产物研究与开发.2000,12(2):27-31
[56]Nanba H.Maitake mushroom immune therapy to prevent from cancer growth and metastasis.Explore.1995,6(1):74-78
[57]Adachi Y,Okazaki M,Ohno N,Yadomae T.Enhancement of cytokme production by macrophages stimulated with(1->3)-β-D-Glucan,Grifolan(GRN).isolated from Grifola frondosa.Biological and Pharmaceutical Bulletin.1994.17(12):1554-156026.
[58]Adachi Y,Ohno N,Yadomae T.Activation of kupffer cells by administration with gel-forming (1-3)-β-D-Glucan from Grifola frondosa.Biological and Pharmaceutical Bulletin.1998.21(3):278-283
[59]Atsuyuki Inoue,Noriko Kodama,Hiroaki Nanba.Effect of Maitake(Grifola frondosa D-Fraction on the T Lymph Node Th-1/Th2 Proportion.Biol.Pharm.Bull.2002,25(4):536-540
[60]Hong Lina,Yu-Hong Sheb,Barrie R et al.Maitake beta-glucan MD-fraction enhances bone marrow colony formation and reduces doxorubicin toxicity in vitro.International Immunopharmacol ogy.2004,4:91-99
[61]邢增涛,周昌艳,潘迎捷等.灰树花多糖研究进展.食用菌学报.1999.6(3):54-58
[62]Ooi VEC.Hepato protective effect of some edible mushrooms.Phytotherapy Research,1996,10(6):536-538
[63]Chang-Qing Gu,Jim-Wen Li,Fu-Huan Chao.Inhibition of hepatitis B virus by D-fraction from Grifola frondosa:Synergistic effect of combination with interferon-(?)in HepG2 2.2.15.Antiviral Research.2006,72:162-165
[64]Kubo K,Aoki H,Nanba H.Anti-diabetic activity present in the fruit body of Griola frondosa.Biological and Pharmaceutical Bulletin.1994,17(8):1106-1110
[65]Kubo K,Nanba H.Anti-hyperliposis effect of Maitake fruit body(Grifola frondosa).Biological and Pharmaceutical Bulletin.1997,20(7):781-785
[66]Nanba H,Adachi Y,Otsuka M et al.Blood pressure-lowering activity present in the fruit body of Grifola frondosa.Chemical and Pharmaceutical Bulletin.1994,36(3):1000-1006
[67]Kazunori S,Noriko K,Hiroaki N.Effect of Maitake(Grifola frondsa)Polysaccharide on Collagen-induced Arthritis in Mice.Jpn.J.Pharmacol.2000,84:293-300
[68]李志州,邓百万,杨海涛,陈文强.美味牛肝菌多糖中β-葡聚糖的含量测定[J].食品科学,2006,27:190-192.
[69]李志州,邓百万,杨海涛.大枣多糖中β-葡聚糖的含量测定[J].食品与发酵工程,2005,31(2):117-119.
[70]何荣军,孙培龙,高红林等.姬松茸多糖中β-葡聚糖含量的酶法测定[J].食品与发酵工业,2004.30(1):124-126.
[71]连喜军,鲁晓翔,蔡保松.酶法测定大麦中β-葡聚糖[J].粮油与油脂,2006,5:12-15.
[72]熊茉君,王新,张红霞.β-葡聚糖含量测定方法的比较[J].华西药学杂志,2005,20(5):438-440
[73]鱼仲爀,朴夫洙,朱昌宪等.用于检测1,3-β-葡聚糖组合物,其制备方法和检测β-葡聚糖的诊断试剂盒.中国专利,01803983.9,2003-03-26.
[74]Finkelman M.Glucatell-a new kit for quantitative determination of 1,3-β-glucan[J].LAL update,2001,19:1-3
[75]杨晓彤,Jennifer M.F.WAN,糜可,冯慧琴,Daniel K.O.CHAN,杨庆尧.鲎G因子实验法定量测定药用、食用蘑菇多糖中的(1,3)-β-葡聚糖含量[J].菌物系统,2003,22(2):296-302。
[76]高冈文,土谷正和,川边惠子等.测量含于样品中的内毒素或1,3-β-D-葡聚糖的方法及其试剂.中国专利,95118546.2,2002-3-20.
[77]闻平.血清内毒素β-1,3-D-葡聚糖检测方法的探讨[J].上海医学检验杂志,2001,16(2):96-97.
[78]Lisa M.Graham,S.Vicky Tsoni,Janet A.Willment,et al.Soluble Dectin-1 as a tool to detect β-glucans[J].Journal of Immunological Methods,2006,314:164-169.
[79]Brown,GD,Herre,J.,Williams,DL,Willment,JA,Marshall,AS,Gordon,S.Dectin-1Mediates the Biological Effects of β-Glucans[J].The Journal of Experimental Medicine,2003,197:1119-1124.
[80]Sanghoon Kim,George E.Inglett.Molecular weight and ionic strength dependence of fluorescence intensity of the Calcofluor/β-glucan complex in flow-injection analysis[J].Journal of Food Composition and Analysis,2006,19:466-472.
[81]Wood,P.J,Fulcher,R.G.Specific interaction of aniline blue with(1-3)-β-D-glucan[J].Carbohydr.Polym,1984,4:49-72.
[82]Yuan- Tin Ko,Yuling Lin.1,3-β-glucan Quantification by a Fluorescence Microassay and Analysis of Distribution in Foods[J].Journal of Agriculture and Food Chemistry,2004.52:3313-3318.
[83]Evans,N.A,Hoyne,P.A,Stone,B.A.Characteristics and specificity of the interaction of a fluorochrome from aniline blue(sirofluor)with polysaccharides[J].Carbohydr.Polymer.1984,4:215-230.
[84]连喜军,鲁晓翔.林旭辉.苯酚-硫酸法测定β-葡聚糖含量的研究[J].粮食与油脂,2005. 10:39-43.
[85]连喜军,鲁晓祥,刘勤生.荧光素纳法测定β-葡聚糖的研究[J].粮油与食品,2006.9:24-26.
[86]汪海波,刘大川,谢笔钧.改进荧光法测定β-葡聚糖含量研究[J].中国粮油学报.2004,1(19):70-74.
[87]李玲.多糖的分离、纯化与分析[J].新疆工学院学报,1996,17(2):129-132.
[88]孙群,牟世芬,陆德培.单糖和寡糖的离子色谱法分析研究[J].化学通报,1991,8(3):39-41.
[89]李润秋.针裂蹄多糖的研究[J].药学学报,1983,18(6):430.
[90]董群,方积年.寡糖及多糖甲基化的发展及现状[J].天然产物开发与研究,1995,7(2):60-65.
[91]方一苇.具有药理活性多糖的研究现状[J].分析化学,1994,22(9):955-960.
[92]张俐娜,丁琼,张平义,等.茯苓菌核多糖的分离和结构分析[J].高等学校化学学报,1997.18(6):990-993.
[93]Grigorij K,Dusan U,Jean R B,et al.Structural and immunochemical characterization of the type Ⅲ group B Streptococcus capsular polysaccharide[J].J Biochem,1996,271(16):8786-8790.
[94]霍光华,李来生.波谱学在多糖结构分析中的应用[J].生命的化学,2002,22(2):194-196.
[95]Agrawal P K.NMR spectroscopy in the structure elucifation of oligosaccharides and glycosides[J].Phytochemistry,1992,31(10):3307-3330.
[96]苏克曼,潘铁英,张玉兰.波谱解析法[M],上海:华东理工大学出版社,2002,8.
[97]沈其丰.核磁共振碳谱[M].北京:北京大学出版社,1988.10.
[98]杨立.二维核磁共振简明原理及图谱解析[M].兰州:兰州大学出版社.
[99]Elisabeth J F,Daan J V,Johannis P K,et al.Characterization of the exopolysaccharide produced by Streptococcus thermophilus 85 containing an open chain nonionic acid[J].Eur J Biochem,2002,269:5590-5598.
[100]Niklas B G.Balakrish N,Andrej W,et al.Structure of the O-Polysaccharide from Vibrio cholerae O6[J].Carbohydr Res,2002,337:813-817.
[101]Oussama A,Alicia P,Juan A L,et al.Fungal cell wall galactomanman isolated from Apodus deciduus[J].Carbohydr Res,2002,337:1503-1506.
[102]Dominaika D,Philip V T,Nikolay P A,et al.Structure the O-specific polysaccharide of Proteus penneri 103 containing ribitol and 2-aminoethanol phosphates[J].Carbohydr Res,2002,337:1535-1540.
[103]Anna N K,Sofa'ya N S,Andrei I G,et al.Structure of the O-polysaccharide of Proteus mirabilis O38 containing 2-acetamidoethyl phosphate and N-linked D-aspartic acid[J].Carbohydr Res,2003,338:2387-2392.
[104]Nikolay P,David B,Minnie R,et al.Structure analysis of the polysaccharide from the lipopolysaccharide of Porphyromonas gingivalis strain W50[J].Eur J Biochem,2001.268:4698-4707.
[105]Wojciech J,Jolanta C,Tomasz N,et al.Structure studies of the O-Specific polysaccharide of Hafnia alvei strain PCM 1207 lipopolysaccharide[J].Eur J Biochem,1999,266:53-61.
[106]Grigorij K,Dusan U,Jean R B,et al.Structural and Immunochemical Characterization of the Type Ⅷ Group B Streptococcus Capsular Polysaccharide[J].J Biochem,1996,271(16):8786-8790.
[107]Mikael S,Felipe U,Andrej W,et al.Structural elucidation of the O-antigenic polysaccharides from the enteroaggregative Escherichia coli strain 62D_1[J].Eur J Biochem,1999,262:56-62.
[108]Prabhakar G R,Unaiza H,Xu Q W,et al.Structure determination of the capsular polysaccharide from Vibrio vulnificus strain 6353[J].Eur J Biochem,1998,255:279-288.
[109]Wang Y,Wiltrud M K,Michael H R,et al.Structural basis of the abscess modulating polysaccharide A2 from Bacterodies fragilis[J].PNAS,2000,97(25):13478-13483.
[110]Leslie M R,Haralambos P,Lesley A S P.The structure of the O-specific Polysaccharide of Escherichia coli O117:K98:H4[J].Carbohydr Res,2000,323:103-110.
[111]A.Molinaro,Vincezo Piscopo,Rosa Lanzetta,et al.Structural determination of the complex exopolysaccharide from the virulent strain of Cryphonectria parasitica[J].Carbohydr Res,2002,337:1707-1713.
[112]Sof'ya N S,Yuriy A K,Alexander S S,et al.Structure of the O-polysaccharide of Erwinia carotovora ssp.Carotovora GDPB436[J].Carbohydr Res.2003,338:2025-2027.
[113]Leslie M R,Haralambos P,Lesley A S P.The structure of the O-antigen of Escherichia coli O116:K+:H10[J].Carbohydr Res,1999,323:246-256.
[114]Come J M S,Xu Q W,Mary R,et al.Structural analysis and chemical depolymerization of the capsular polysaccharide of Streptococcus pneumoniae type 1[J].Carbohydr Res,2001.337:335-344.
[115]Helene R,Nelly K,Roger P,et al.Structure of the exopolysaccharide of Vibrio diabolicus isolate from a deep sea hydrothermal vent[J].Carbohydr Res,1999,322:40-45.
[116]Krystyna Z,Filip V T,Nikolay P A,et al.Structure of the O-specific polysaccharide of Proteus mirabilis D52 and typing of this strain to Proteus serogroup O33[J].Eur J Biochem.2001,268:4346-4351.
[117]Clas L,Andrej W,Goran W.Structural analysis of the O-antigen polysaccharide from the shiga toxin-producing Escherichia coli O172[J].Eur J Biochem,2001,268:2239-2245.
[118]张维杰.糖复合多糖生化研究技术[M].第二版.浙江:浙江大学出版社,1994.
[119]魏远安,方积年.高效凝胶渗透色谱法测定多糖纯度及分子量[J].药学学报,1989,24:532-536.
[120]Liu,C P,Bao,X F,Fang,J N.Retention behaviors of uronic acid-containing polysaccharides in HPGPC[J].Chin Chem Lett,2001,12:909-912.
[118]李玲.多糖的分离、纯化与分析[J]_新疆工学院学报,1996,17(2):129~132.
[121]Staaf M,Urbina F,Weintraub A,WidmalmL G.Structural elucidation of the O-antigenic polysaccharides from the enteroaggregative Escherichia coli strain 62D_1.European Journal of Biochemistry,1999,262:56~62.
[122]黄年来.自修食用菌学[M].南京:南京大学出版社,1986.
[123]徐桂云,陈汝贤,常理文.用毛细管气相色谱法测定多糖中单糖的组成[J].分析测试学报.2000,19(3):71-73.