火棘多糖的初步研究
详细信息    本馆镜像全文|  推荐本文 |  |   获取CNKI官网全文
摘要
火棘(Pyracantha fortuneana(Maxim.)Li),又叫赤阳子、救军粮,主要分布于我国东南和西南部。其果实、叶和根都可药用,果实具有健脾消积、活血止血等功能。同时也可食用,酸甜适口、香气浓郁,含有丰富的营养物质。因此作为药食两用的火棘具有很大的开发价值。
     为了进一步开发火棘野生资源,本论文对秦巴山区产火棘果实中水溶性多糖的提取、分离纯化、理化性质、生物活性、膳食纤维的制备及特性等方面进行了较为深入的研究,取得了如下的研究结果。
     火棘水溶性多糖的最佳提取工艺为:90℃水,提取9小时,料液比为1:10;在此条件下火棘多糖的提取率约为3.5%;超声波处理可使火棘多糖的得率提高30%以上。
     采用DPPH·分光光度法对火棘粗多糖进行体外抗氧化试验,结果表明火棘粗多糖具有明显的清除DPPH·作用,SC_(50)为1.87mg/mL。
     经试验,蛋白酶-Sevag法具有较好的脱蛋白效果;LSA-8大孔吸附树脂可以用于火棘粗多糖的脱色;再经DE-52和Sephadex G-200柱层析分离得到PP-A2、PP-A3、PP-B2三种多糖组分。
     气相色谱、红外光谱、理化特性分析结果表明PP-A2和PP-B2是同一物质,主要由阿拉伯糖和鼠李糖组成,其中还含有少量的果糖、半乳糖和甘露糖,阿拉伯糖:鼠李糖:果糖:半乳糖:甘露糖=5.05:4.51:3.37:1.88:1,经Sephadex G-200柱层析测定其分子量为30,000Dal,不同的是PP-B2中含有少量多酚类色素物质。PP-A3是一种糖蛋白,分子量为210,000Dal,主要由阿拉伯糖组成,其中含有少量的葡萄糖和果糖,阿拉伯糖:葡萄糖:果糖=1.62:1:1.59,其中氨基酸总含量为0.5%(半胱氨酸未检出)。
     火棘多糖作为膳食纤维类功能性食品添加剂,具有较好的膨胀力、持水力及阳离子交换能力,同时具有较好的清除自由基能力,其作为面条改良剂时的最适使用量为3%左右。
Abstract Pyracantha fortuneana (Maxim.) Li, which was also named Chiyangzi and Jiujunliang, is mainly distributed in the southeast and southwest of China. Its fruit, leaf and root all could be used as medicine in China. Furthermore,its fruit was edible, which contained abundant nutriment. So Pyracantha fortuneana (Maxim.) Li fruit had great exploitation value.
    In order to ulteriorly exploit Pyracantha fortuneana (Maxim.) Li resource, the author conducted a series of study on the water-soluble polysaccharide from Pyracantha fortuneana and achieved the following results.
    The optimal extraction technology of Pyracantha fortuneana polysaccharides were 90℃, 9h, 10:1 (WAV) of water to its fruit. Extraction ratio of the polysaccharides was about 3.5%. With ultrasonic, extraction ratio can be increased more than 30%.
    Through antioxidant in vio experiment by DPPH . spectrophotophy, it showed that Pyracantha fortuneana crude polysaccharide could remove DPPH . and its effect was very prominent and SC50 was 1.87mg/mL.
    Some experiments indicated that the protein could be almost clearly removed from Pyracantha fortuneana crude polysaccharide by protein-enzyme-Sevag and decolor could be achieved by LSA-8 big size adsorption resin. Then isolated by DE-52 and Sephadex G-200 gel filtration, three glycoconjugates PP-A2,PP-A3 and PP-B2 were attained.
    Gas chromatography analysis, infra-red spectra analysis and physical and chemical characters showed that PP-A2 and PP-B2 were the same materials. They mainly contained rhamnose, arabinose, fructose, galactose and mannose in a ratio of 5.05:4.51:3.37:1.18:1. Sephadex G-200 gel filtration analysis showed that their molecular weight was 30,000dal. Different to PP-A2, PP-B2 contained a little pigment. PP-A3 was a kind of glycoprotein. It was composed of arabinose, glucose and fructose in a ratio of 1.62:1:1.59 and some amino acids whose total content was 0.5%. In amino acid, cysteine was not detected out. The molecular weight of PP-A3 was 210,000dal.
    As dietary fiber functional food additivies, Pyracantha fortuneana polysaccharide has good expansibility, ability to keep water and ability to exchange with positive ion. Furthermore, it can scavenge free radical. When it is used as meliorative additive in noodle, the dosage of Pyracantha fortuneana polysaccharide is about 3%.
引文
[1] 崔亦华,崔英德,易国斌.应用广泛的天然多糖及其提取方法.广州化工,2002,3(30):7~9
    [2] Sugiura Met al. Methods in carbohydrate extraction from Fungi, Japan J Pharmacol, 1980,30:506~508
    [3] 方积年.多糖的分离纯化及其纯度鉴别、分子量测定.药学通报,1984,19:622~625
    [4] Fukuda K et al. The extraction of remella fuciformis polysaccharides by enzymatic technique, Chem.Anal.Ed., 1982,23: 1955~1958
    [5] 张翼伸,李治平等,云芝多糖的分离与鉴定,吉林师范大学学报(自然科学版),1979,2:108~113
    [6] 张翼伸.多糖的结构测定.生物化学与生物物理进展,1983,53:18~23
    [7] 王仲俘.博士学位论文
    [8] 董群,方积年.寡糖及多糖甲基化方法的发展现状.天然产物研究与开发,1995,7(2):60~64
    [9] 张惟杰.糖复合物生化研究技术.浙江大学出版社,1994:158
    [10] Bengt L,Jorgen U Methylation analysis of complex carbohydrates:general procedure and application for sequence analysia. Methods Enzymol, 1978, 50: 3~33
    [11] 张国文,陈梦菁.多糖的甲基化方法.化学通报,1988,11:41~42
    [12] 方积年.多糖研究的现状.药学学报,1986,21(12):944~950
    [13] Agawal P K, Pathak A K. Nuclear magnetic resonance spectroscopic approaches for the determination of interglycosidic linkage and sequence in oligosaccharides. Phytochmical Analysis, 1996,7:113~130
    [14] Agawal P K. NMR spectroscopy in the structural elucidation of oligosaccharides and glycosides. Phytochemistry, 1992,31 (10):3307~3330
    [15] 胡亚南,王义明,罗国安.质谱在糖分析中的应用.药学学报,2000,35(5):397~400
    [16] 方一苇.快原子轰击质谱研究寡糖及其缀合物结构的进展.分析化学,1980,18(11):1072~1077
    [17] Egge H, Peter-Katalinic J. Fast atom bombardment mass spectrometry fpr structural elucidation of glycoconjugate. Mass Spectrometry Reviews, 1987, 6(2):
    
    331~393
    [18] Reinhold V N, Carr S A. New mass spectral approaches to complex carbohydrate structure. Mass Spectrometry Reviews, 1983,2(2): 153~221
    [19] Suzuki S, Kakehi K, Honda S. Comparision of the sensitivities of various derivatives of oligosaccharides in LC/MS with fast bombardment and electrospray ionization interfaces. Analytical Chemistry, 1996,68(13):2073~2083
    [20] Duffin K L, Lange, Gary W, et al. Identification and oligosaccharide structure analysis of rhodopsin glycoforms containing galactose and sialic acid. Glycobiology, 1993, 3 (4):365~380
    [21] Fenn J B, Mann M, Meng C K, et al. Electrospray ionization for mass spectrometry of large biomolecular. Science, 1989,246:64~71
    [22] Garozza D, Impallomen G, Spina E, et al. Linkage analysis in disaccharides by electrospray mass spectrometry. Carbohydrate Research, 1991,221:253~257
    [23] Yoshimi Y, Yamazaki S, Ikekita M. Developmental changes in Asn-linked neutral oligosaccharides in murine cerebrum. Biochim. Biophys. Acta-General Subjects, 1999,142691):69~79
    [24] Weiskopf A S, Vouros P, Harvey D J. Electrospray ionization-ion trap mass spectrometry for structural analysis of complex N-linked glycoprotein oligosaccharides. Analytical Chemistry, 1998,70(20):4441~4447
    [25] Mock K K, Davey m, Cottrell J S. The analysis ofunderivatized oligosaccharides by matrix-assisted laser desorption mass spectrometry. Biochem, Biophys, Res, Comm, 1991,177(2):644~651
    [26] Stahl B, Steup M, Karas M, et al. Analysis of neutral oligosaccharides by Matrix-assisted laser desorption/ionization mass spectrometry. Analytical Chemistry, 1991, 63(14): 1463~1466
    [27] Yamamoto k. Microbial endoglycosidases for analysis of oligosaccharide chains in glycoproteins. J Biochem, 1994,116:229~235
    [28] Edge C J,Rademacher T W, Wormald M R, et al. Fast sequencing of olgosaccharides: the reagent-array analysis method. Pro.Natl.Acad.Sci.USA, 1992,89:6388~6342
    [29] 王蓉,吴剑波.多糖生物活性的研究进展.国外医药抗生索分册,2001,22(3):97~100
    [30] 陈洪亮,李伯涛,张家祥等.免疫活性多糖的免疫调节作用及机制研究进展.中国药理学通报,2002,18(1):249~252
    [31] Donzis B A. Solubilized yeast glucan. 1996, United States Patent5519009
    
    
    [32] Sherwood E R, Williams D L, et al. Enhancement of interleukin-1 and interleukin-2 production by soluble glucan. Immunopharmacol, 1987, 9(3): 261~267
    [33] Estrada A, Yun CH, et al. Immunomodulatory activities of oat β-glucan in vitro and in vivo. Microbial immunol, 1997, 41(12):991~998
    [34] 王玲,杜手英.枸杞多糖的免疫调节研究进展.上海免疫学杂志,1995,15(2):118~120
    [35] 李金锋,郭静文等.猪苓多糖、香菇多糖和分枝杆菌多糖对淋巴因子活化杀伤细胞活性增强作用的研究.中国中西医结合杂志,1996,16(4):224~226
    [36] 田庚元.中药免疫调节剂的研究和开发.99中国药学会学术年会论文集.1999,8:570~575
    [37] 黄芳,蒙义文.活性多糖的研究进展.天然产物研究与开发.1999,11(6):90~98
    [38] Yamada H, Ohtani K, Kiyohara H, et al. Purification and chemical properties of anti-complementary polysaccharide from the leaves of artemisia princepes. Planta Medic, a, 1985,51:12~127
    [39] 毛俊浩,吕志良,曾群力,阮红.白术多糖对小鼠淋巴细胞功能的调节.免疫学杂志,1996,12(4):233~236
    [40] 阮红.女贞子多糖对小鼠淋巴细胞IL—2诱生的调节作用.上海免疫学杂志,1999,19(6):337
    [41] Kim H M, et al. Stimulation of humoral and cell mediated immunity by polysaecharide from mushroom Phellinus linteus. Immunopharmacol, 1996,18 (5) :295~303
    [42] 林志彬.灵芝多糖的免疫药理学及其研究意义.北京医科大学学报,1992,24(4):271~274
    [43] 耿长山,刑善田,周金黄.黄芪多糖对去T细胞小鼠促进产生机理的探讨.上海免疫学杂志,1985,5(2):69~71
    [44] 田庚元,冯宇澄,林颖.植物多糖的研究进展.中国中药杂志,1995,20(7):441~443
    [45] Stimpel M, et al. Macrophage activation and induction of macrophage cytotoxicity by purified polysaccharide fractions from the plant Eehinacea purpurea. Infect Immunopharmacol, 1984, 46(3):845~849
    
    
    [46] 李祖伦.川牛膝多糖的免疫活性研究.中药材,1998,21(2):90~92
    [47] 张小梅.黄芪多糖的免疫调节作用及抗肿瘤作用研究进展.大连大学学报,2003,12,24(6):101~104
    [48] 宁安红,孙晓东,曹婧,黄敏.灵芝多糖抗肿瘤作用的初步探讨.中国微生态学杂志,2003,12,15(6):335~336
    [49] 陈留勇,孟宪军,贾薇等.黄桃水溶性多糖的抗肿瘤作用及清除自由基、提高免疫活性研究.2004,25(1):167~170
    [50] 崔文,尹苗,安利国.马齿苋多糖的抗肿瘤活性.山东师大学报(自然科学版)2002,3,17(1):73~76
    [51] 周林珠,杨祥良,周井炎,徐辉碧.多糖抗氧化作用研究进展.中国生化药物杂志,2002,23(4):210~212
    [52] 何书英,詹彤,王淑如.山药水溶性多糖的化学及体外抗氧化活性.中国药科大学学报,1994,25(6):369~372
    [53] 张振涛,吴泉,吴仁奇,沈传勇.茜草多糖的抗氧化作用.内蒙古医学院学报,1998,3,20(1):31~33
    [54] 苗明三.百合多糖抗氧化作用研究.中药药理与临床,2001,17(2):12~13
    [55] Yeung J H, Chiu L C. Effect of Polysaccharide peptide(PSP) on glutathione and protection against paracetamol induced hepatotoxicity in the rat. Methods Find Exp Clin Pharmacol, 1994,16(10):723
    [56] 黄志江,季晖等.人工虫草多糖降血糖作用及其机制研究.中国药科大学学报,2002,33(1):51~54
    [57] 刘成梅,付桂明等.百合多糖降血糖功能研究.食品科学,2002,23(6):113~114
    [58] 张拥军,姚惠源.南瓜多糖的分离、纯化及其降血糖作用.中国粮油学报,2002,8,17(4):59~62
    [59] 陈建国,王茵,梅松等.茶多糖降血糖、改善糖尿病症状作用的研究.营养学报,2003,25(3):253~255
    [60] 左绍远,万顺康等.钝项螺旋藻多糖降血糖调血脂实验研究.中国生化药物杂志,2000,21(6):289~291
    [61] 王长云,管华诗.多糖抗病毒作用研究进展.生物工程进展,2000,20(1):18~20
    
    
    [62] 项哨,朱圣禾.灰树花多糖在小鼠体内抗病毒作用的研究.浙江医科大学学报,1995,24(5):203~205
    [63] 陈玉香,张丽萍,梁忠岩等.沙棘果水溶性多糖Hn的分离纯化与抗病毒研究.东北师大学报自然科学版,1997,4:74~77
    [64] 王长云,管华诗等.多糖抗病毒作用研究进展Ⅱ.硫酸多糖抗病毒作用.生物工程进展,2000,20(2):3~8
    [65] 田庚元,李寿桐等.牛膝多糖硫酸酯的合成及其抗病毒活性.药学学报,1995,30(2):107~111
    [66] Tu NTN, Chinh LH. Investigation of the antibacterial effect of a natural polysaccharide. Tap Duoc Hoc, 1997,10:17
    [67] 方一苇.具有药理活性多糖的研究现状.分析化学,1994,22(9):955
    [68] 宋丽艳,马文霞等.银杏细胞培养物多糖和银杏叶多糖生物活性的研究.中国生化药物杂志,1999,20(6):278
    [69] 吴倩,吴剑波,李元.白细胞介素—1受体拮抗剂139A的理化性质及体内活性研究.中国抗生索杂志,1999,24(6):401
    [70] 中国野生果树
    [71] 王军宪,牛娟芳,尤晓娟.秦岭产火棘果实的成分分析.西北药学杂志,1994,10,9(5):204~206
    [72] 侯建军,魏文科,薛慧,张宏.野生植物火棘的研究进展.湖北民族学院学报(自然科学版),2002,20(1):15~18
    [73] 王和声,柴立,李守兰.赤阳子对老龄大鼠机体SOD及LPO的影响.中草药,1997,28(3):158~161
    [74] 安性和,林开颜,陆林杰.赤阳子降血脂及防止动脉硬化斑块形成的实验研究.贵州医药,2000,24(4):254~255
    [75] 侯建军,刘希林,武模戈.野生植物火棘抗疲劳的功效研究.武汉科技学院学报,2002,15(5):66~68
    [76] 侯建军,刘希林等.火棘消食健脾的动物试验.湖北农业科学,2003,4:84~86
    [77] 向纪明.火棘红色素的提取及稳定性研究.安康师专学报,1997,1:42~44
    [78] 鲁周民等.火棘饮料综合加工技术.中国林副特产,1999(2):39~40
    [79] 丁筑红.绿豆芽火棘复合饮料的研制.食品研究与开发,1999,20(2):32~35
    
    
    [80] 邓如福,王三根.火棘饮料酿制工艺技术研究.中国野生植物,1989,1:34~37
    [81] 洪若芬等.火棘酒的制造方法.酿酒科技,1999,6:104
    [82] Dubois M. Colorimetric method for determination of sugars and substance[J]. Anal Chem. 1956 (28): 350~356
    [83] Larrauri JA, Sanchez-Moreno C, Saura-Calixto F. Effect of temperature on the free radical scavenging capacity of extracts from red and white grape pomace peels. Journal of Agric Food Chem, 1998,46(7):2694~2697
    [84] Yokozawa T, Dong E, Natagawa T, et al. In vitro and in vivo studies on the radical-scavenging activity of tea. Journal of Agric Food Chem, 1998, 46(6): 2143~2150
    [85] 许申鸿,杭瑚.20种植物水提物对DPPH的清除作用.中国野生植物资源,1999,3:50~51
    [86] 谭仁祥等.植物成分分析[M].北京:科学出版社,2002:76-78.
    [87] 张部昌,吴东儒,陶乐平.天门冬多糖的分离纯化及其部分理化性质.安徽大学学报(自然科学版),1994,2:88~94
    [88] 何书英,詹彤,王淑如.山药水溶性多糖的化学及体外抗氧化活性.中国药科大学学报,1994,6:369~372
    [89] 张林生,蒋纪云,张宝军.小麦籽粒发育过程中氨基酸的变化.作物学报,1997,5:302~306
    [90] Parker FS.Application of infrared, raman and resonance spectroscopy. Acdemic Press. New York 1982:52~56
    [91] S.Suzarme Nidsen著,杨严俊等译.食品分析.中国轻工业出版社,2002,7
    [92] 金茂国,孙伟.挤压对豆渣膳食纤维理化性质影响.粮食与饲料工业,1996,11:35~38
    [93] 郑建仙,耿立萍,高孔荣.蔗渣膳食纤维挤压改性的研究(Ⅱ).华南理工大学学报(自然科学版),1996,12:64~68

© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号

地址:北京市海淀区学院路29号 邮编:100083

电话:办公室:(+86 10)66554848;文献借阅、咨询服务、科技查新:66554700