羊栖菜褐藻糖胶的结构表征及其抗氧化活性
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摘要
为研究羊栖菜褐藻糖胶的结构及其抗氧化活性,以羊栖菜为原料,经CaCl_2溶液提取、DEAE-Sepharose fast flow分离纯化得到褐藻糖胶组分SFP-2。采用高效凝胶渗透色谱法(HPGPC)、化学法、高效液相法(HPLC)以及傅里叶变换红外光谱法(IR)对多糖的纯度、分子量、理化性质、单糖组成以及官能团进行测定,并通过测定DPPH自由基清除活性、超氧阴离子清除活性、ABTS~+自由基清除活性、亚硝酸盐清除活性和还原力,对多糖的体外抗氧化活性进行了评价。结果表明,SFP-2分子量均一,为707.0 kDa;总糖含量为66.9%,蛋白含量为3.1%,硫酸基含量为21.2%,不含糖醛酸; SFP-2主要由岩藻糖和半乳糖构成,还含有少量的甘露糖和氨基葡萄糖;红外光谱显示,SFP-2具有硫酸化多糖常见官能团的特征吸收峰;上述结果表明SFP-2是一种不含糖醛酸的硫酸化岩藻聚糖。SFP-2具有良好的清除DPPH自由基、超氧阴离子、ABTS+自由基、亚硝酸盐活性,其EC50分别为8.94、8.30、16.04、9.87 mg/mL,且具有一定的还原能力。因此,SFP-2是一种不含糖醛酸的,抗氧化活性良好的羊栖菜褐藻糖胶。
In order to study the structural characterization and antioxidant activities of fucoidan from Sargassum fusiforme,a heterogeneous polysaccharide SFP-2 was obtained by CaCl_2 solution extraction and anion-exchange.Structural characterization of SFP-2 was preliminarily determined by high performance gel permeation chromatography( HPGPC),chemical methods,high performance liquid chromatography(HPLC) as well as fourier-tansform infrared(IR).The antioxidant activities were evaluated by scavenging assays involving DPPH radical,superoxide anion,ABTS~+ radical,nitrite and reducing power. Results indicated that SFP-2 appeared as a single and symmetrical peak in the HPGPC chromatogram and the average molecular weight was 707.0 kDa.The contents of total sugar,protein and sulfate ester were 66.9%,3.1% and 21.2%,respectively.No uronic acid was detected,SFP-2 was mainly composed of fucose and galactose with a small amount of mannose and glucosamine. IR further proved SFP-2 was a sulfated polysaccharide.From the results,SFP-2 was a sulfated fucoidan without uronic acid.SFP-2 had good scavenging activity for DPPH radical,superoxide anio,ABTS~+ radical and nitrite.The EC_(50) values of the scavenging assays were 8.94,8.30,16.04,9.87 mg/mL,respectively,with the ability of reducing power.Results above indicated that SFP-2 was a kind of non-glucuronic acid fucoidan which had better antioxidant activity.
In order to study the structural characterization and antioxidant activities of fucoidan from Sargassum fusiforme,a heterogeneous polysaccharide SFP-2 was obtained by CaCl_2 solution extraction and anion-exchange.Structural characterization of SFP-2 was preliminarily determined by high performance gel permeation chromatography( HPGPC),chemical methods,high performance liquid chromatography(HPLC) as well as fourier-tansform infrared(IR).The antioxidant activities were evaluated by scavenging assays involving DPPH radical,superoxide anion,ABTS~+ radical,nitrite and reducing power. Results indicated that SFP-2 appeared as a single and symmetrical peak in the HPGPC chromatogram and the average molecular weight was 707.0 kDa.The contents of total sugar,protein and sulfate ester were 66.9%,3.1% and 21.2%,respectively.No uronic acid was detected,SFP-2 was mainly composed of fucose and galactose with a small amount of mannose and glucosamine. IR further proved SFP-2 was a sulfated polysaccharide.From the results,SFP-2 was a sulfated fucoidan without uronic acid.SFP-2 had good scavenging activity for DPPH radical,superoxide anio,ABTS~+ radical and nitrite.The EC_(50) values of the scavenging assays were 8.94,8.30,16.04,9.87 mg/mL,respectively,with the ability of reducing power.Results above indicated that SFP-2 was a kind of non-glucuronic acid fucoidan which had better antioxidant activity.
引文
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[25]李娜,李菁,刘雪,等.抗凝血活性海洋硫酸多糖HP1-1的结构研究[J].中国海洋药物,2015,34(5):7-12.
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[28]林聪,赵春琦,李娜,等.1种绿藻硫酸化多糖的化学组成及其结构表征[J].中国海洋药物,2014,33(4):55-58.
[2]季宇彬,高世勇,张秀娟.羊栖菜多糖体外抗肿瘤作用及其诱导肿瘤细胞凋亡的研究[J].中草药,2003,34(7):638-640.
[3]季宇彬,高世勇.羊栖菜多糖体外抗肿瘤作用及其作用机制的研究[J].中草药,2003,34(12):1111-1114.
[4]王兵,李靖,马舒冰,等.羊栖菜多糖降血压作用的实验研究[J].中国海洋药物,2000(3):33-35.
[5]冯磊,沈健,徐明智.羊栖菜降血糖作用[J].中华预防医学杂志,2001,35(5):324.
[6]宁准梅,王勤,林之川.羊栖菜多糖降血糖、调节血脂效果研究[J].中国食品学报,2003(z1):126-129.
[7]邵平,陈蒙,裴亚萍,等.径向流色谱分离纯化海藻多糖及其抗氧化活性比较分析[J].核农学报,2013,27(5):635-640.
[8]Chizhov A O,Dell A,Morris H R,et al.A study of fucoidan from the brown seaweed Chorda filum[J].Carbohydrate Research,1999,320(2):108-119.
[9]何丹,张旭,肖保衡,等.羊栖菜多糖的提取和抗氧化活性研究[J].海洋科学,2016,40(12):24-29.
[10]丁浩淼,李伟强,汪财生,等.羊栖菜活性组分多糖的体外抗氧化能力比较[J].核农学报,2016,30(5):920-925.
[11]陈琳琳,谢志新,程晶,等.羊栖菜多糖提取物及其润肤霜的保湿和抗氧化活性[J].日用化学工业,2018(1):31-36.
[12]Silchenko A S,Rasin A B,Kusaykin M I,et al.Structure,enzymatic transformation,anticancer activity of fucoidan and sulphated fucooligosaccharides from Sargassum horneri[J].Carbohydrate Polymers,2017,175:654-660.
[13]Yan M X,Mao W J,Chen C L,et al.Structural elucidation of the exopolysaccharide produced by the mangrove fungus Penicillium solitum[J].Carbohydrate Polymers,2014,111:485-491.
[14]Dubois M,Gilles K A,Hamilton J K,et al.Colorimetric method for determination of sugars and related substances[J].Analytical Chemistry,1956,28:350-356.
[15]Zhang X Q,Paul L B,Margaret J K.Measurement of polysaccharides and proteins in biofilm extracellular polymers[J].Water Science and Technology,1998,37(4-5):345-348.
[16]Therho T T,Hartiala K.Method for determination of the sulfate content of glycosaminoglycans[J].Analytical Biochemistry,1971,41:471-476.
[17]Bitter T,Muir H M.A modified uronic acid carbazole reaction[J].Analytical Biochemistry,1962,4:330-334.
[18]Qi X H,Mao W J,Gao Y,et al.Chemical characteristic of an anticoagulant-active sulfated polysaccharide from Enteromorpha clathrata[J].Carbohydrate Polymers,2012,90:1804-1810.
[19]Synytsya A,CˇopíkováJ,Matějka P,et al.Fourier transform Raman and infrared spectroscopy of pectins[J].Carbohydrate Polymers,2003,54:97-106.
[20]Shimada K,Fujikawa K,Yahara K,et al.Antioxidative properties of xanthan on the autoxidation of soybean oil in cyclodextrin emulsion[J].Journal of Agricultural and Food Chemistry,1992,40:945-948.
[21]Marklund S,Marklund G.Involvement of superoxide anion radicals in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase[J].European Journal of Biochemistry,1974,47:469-471.
[22]Re R,Pellegrini N,Proteggente A,et al.Antioxidant activity applying an improved ABTS radical cation decolorization assay[J].Free Radical Biology and Medicine,1999,26(9-10):1231-1237.
[23]赵二劳,梁泽,张海容.沙棘叶对亚硝酸盐清除能力的研究[J].食品工业科技,2006,27(3):81-82.
[24]Dorman H J D,Hiltunen R.Fe(Ⅲ)reductive and free radical-scavenging properties of summer savory(Satureja hortensis L.)extract and subfractions[J].Food Chemistry,2004,88:193-199.
[25]李娜,李菁,刘雪,等.抗凝血活性海洋硫酸多糖HP1-1的结构研究[J].中国海洋药物,2015,34(5):7-12.
[26]Hu P,Li Z X,Chen M C,et al.Structural elucidation and protective role of a polysaccharide from Sargassum fusiforme on ameliorating learning and memory deficiencies in mice[J].Carbohydrate Polymers,2016,139:150-158.
[27]李波,许时婴.羊栖菜褐藻糖胶的分级纯化和结构分析[J].无锡轻工大学学报,2004,23(2):86-93.
[28]林聪,赵春琦,李娜,等.1种绿藻硫酸化多糖的化学组成及其结构表征[J].中国海洋药物,2014,33(4):55-58.