银耳多糖的降解、磷酸酯化修饰及其抗氧化活性
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摘要
目的:磷酸酯化修饰经氧化降解的银耳多糖,并测定修饰多糖的抗氧化活性。方法:采用双氧水-V_C体系降解银耳多糖,并测定其溶解度,以三氯氧磷和吡啶作为磷酸酯化试剂对降解的银耳多糖进行修饰;利用傅里叶红外光谱表征多糖的结构特征;采用磷钼蓝法测定取代度;以羟自由基和DPPH自由基的清除率为指标,比较修饰前后银耳多糖的抗氧化能力。结果:银耳多糖经降解、透析后获得一个单一组分,相对分子质量为7.82 kDa,红外光谱分析结果表明,修饰后的银耳多糖在1262.22、540.55 cm~(-1)处有磷酸酯键生成,磷酸酯化银耳多糖的取代度为0.021,水溶解度为42.71±1.67 mg/mL;修饰后的银耳多糖对羟自由基和DPPH自由基的清除作用具有明显的剂量效应,浓度为2.4 mg/mL时活性最高,对DPPH清除率达到73.77%,对羟自由基清除效率达到67.45%。结论:氧化降解提高了银耳多糖的水溶解度,磷酸酯化修饰增强了银耳多糖的抗氧化能力。
Objective:To modify the oxidized degraded polysaccharides of Tremella fuciformis by phosphate esterification and determine the antioxidant activity of the modified polysaccharides. Methods:The polysaccharides of Tremella fuciformis were degraded by hydrogen peroxide-V_Csystem and its solubility was determined. The degraded Tremella polysaccharides were modified by phosphorus oxychloride and pyridine as phosphorylating reagents. The structural characteristics of polysaccharides were characterized by Fourier transform infrared spectroscopy. The substitution was determined by phosphorus molybdenum blue method. The antioxidant activities of Tremella polysaccharides before and after modification were compared with the scavenging rate of hydroxyl radicals and DPPH free radicals. Results:The Tremella polysaccharides were degraded and dialyzed to obtain a single component with a relative molecular mass of 7.82 kDa. The results of infrared spectroscopy indicated that the modified Tremella polysaccharides had phosphate bond formation at 1262.22、540.55 cm~(-1). The degree of substitution was 0.021.Water solubility was 42.71±1.67 mg/mL. The modified Tremella polysaccharide had a significant dose effect on the scavenging effects of hydroxyl radicals and DPPH free radicals. The activity was highest at the concentration of 2.4 mg/mL,and the DPPH clearance rate of the modified Tremella polysaccharides reached 73.77%. The scavenging efficiency of hydroxyl radicals reached 67.45%. Conclusion:Oxidative degradation enhanced the water solubility of Tremella polysaccharides,and the phosphorylation modification enhanced the antioxidant capacity of Tremella polysaccharides.
Objective:To modify the oxidized degraded polysaccharides of Tremella fuciformis by phosphate esterification and determine the antioxidant activity of the modified polysaccharides. Methods:The polysaccharides of Tremella fuciformis were degraded by hydrogen peroxide-V_Csystem and its solubility was determined. The degraded Tremella polysaccharides were modified by phosphorus oxychloride and pyridine as phosphorylating reagents. The structural characteristics of polysaccharides were characterized by Fourier transform infrared spectroscopy. The substitution was determined by phosphorus molybdenum blue method. The antioxidant activities of Tremella polysaccharides before and after modification were compared with the scavenging rate of hydroxyl radicals and DPPH free radicals. Results:The Tremella polysaccharides were degraded and dialyzed to obtain a single component with a relative molecular mass of 7.82 kDa. The results of infrared spectroscopy indicated that the modified Tremella polysaccharides had phosphate bond formation at 1262.22、540.55 cm~(-1). The degree of substitution was 0.021.Water solubility was 42.71±1.67 mg/mL. The modified Tremella polysaccharide had a significant dose effect on the scavenging effects of hydroxyl radicals and DPPH free radicals. The activity was highest at the concentration of 2.4 mg/mL,and the DPPH clearance rate of the modified Tremella polysaccharides reached 73.77%. The scavenging efficiency of hydroxyl radicals reached 67.45%. Conclusion:Oxidative degradation enhanced the water solubility of Tremella polysaccharides,and the phosphorylation modification enhanced the antioxidant capacity of Tremella polysaccharides.
引文
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[14]贾红倩,刘嵬,梁立,等.红曲霉菌胞外多糖的分离纯化、结构鉴定及抗氧化活性测定[J].食品工业科技,2017,38(12):92-96.
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[16]袁爱群,陶萍芳,赵凤英,等.磷钼蓝法测定磷酸锌中的磷含量[J].分析测试技术与仪器,2004,10(4):251-253.
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[21]周林珠,杨祥良,周井炎,等.多糖抗氧化作用研究进展[J].中国生化药物杂志,2002,23(4):210-212.
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[23]Yang S,Guo Z,Miao F,et al.The hydroxyl radical scavenging activity of chitosan,hyaluronan,starch and their O-carboxymethylated derivatives[J].Carbohydrate Polymers,2010,82(4):1043-1045.
[24]Xu Y,Niu X,Liu N,et al.Characterization,antioxidant and hypoglycemic activities of degraded polysaccharides from blackcurrant(Ribes nigrum,L.)fruits[J].Food Chemistry,2017:S0308814617315832.
[25]杨文鸽,谢果凰,徐大伦,等.龙须菜多糖的降解及其降解产物的抗氧化活性[J].水产学报,2009,33(2):342-347.
[26]宋见喜,任婷,王贺,等.高山红景天多糖的硫酸化修饰及DPPH自由基清除活性研究[J].中草药,2017,48(24):5125-5129.
[27]Kocharova N A,Thomas-Oates J E,Knirel Y A,et al.The structure of the O-specific polysaccharide of Citrobacter O16 containing glycerol phosphate[J].Febs Journal,2010,219(1-2):653-661.
[2]颜军,徐光域,郭晓强,等.银耳粗多糖的纯化及抗氧化活性研究[J].食品科学,2005,26(9):169-172.
[3]聂伟,张永祥,周金黄.银耳多糖的药理学研究概况[J].中药药理与临床,2000,16(4):44-46.
[4]王晓璐,汲晨锋.多糖修饰物及其抗肿瘤作用机制研究[J].氨基酸和生物资源,2015,37(4):12-17.
[5]陈蕾,吴皓.多糖降解方法的研究进展[J].中华中医药学刊,2008,26(1):133-135.
[6]Jin M,Wang Y,Huang M,et al.Sulphation can enhance the antioxidant activity of polysaccharides produced by Enterobacter cloacae Z0206[J].Carbohydre Polymers,2014,99(1):624-629.
[7]Peng Z,Mingyong X,Boqiang F U.A review of the studies on the polysaccharide structure[J].Journal of Nanchang University,2001.
[8]陈晓明,田庚元.多糖磷酸酯化的研究进展[J].有机化学,2002,22(11):835-839.
[9]宁苑灵.磷酸酯化和不同分子量片断龙眼肉多糖的制备及其免疫调节作用和抗氧化作用的研究[D].南宁:广西医科大学,2013.
[10]孙婕,尹国友,牛美艳.磷酸酯化南瓜多糖的制备及抗氧化活性研究[J].四川农业大学学报,2013,31(3):322-327.
[11]Chen B.Optimization of extraction of Tremella fuciformis polysaccharides and its antioxidant and antitumour activities in vitro[J].Carbohydrate Polymers,2010,81(2):420-424.
[12]Zhang Z,Wang X,Zhao M,et al.Free-radical degradation byFe2+/VC/H2O2 and antioxidant activity of polysaccharide from Tremella fuciformis[J].Carbohydrate Polymers,2014,112(21):578-582.
[13]王凌,孙利芹.绿色巴夫藻多糖及降解产品的抗氧化和保湿性能[J].食品科学,2012,33(21):87-90.
[14]贾红倩,刘嵬,梁立,等.红曲霉菌胞外多糖的分离纯化、结构鉴定及抗氧化活性测定[J].食品工业科技,2017,38(12):92-96.
[15]Duan K,Shan H,Lin Y T,et al.Degradation of polysaccharide from Enteromorpha prolifera with hydrochloric acid and hydrogen peroxide assisted by microwave and its antioxidant activity[J].Food Science & Technology,2015.
[16]袁爱群,陶萍芳,赵凤英,等.磷钼蓝法测定磷酸锌中的磷含量[J].分析测试技术与仪器,2004,10(4):251-253.
[17]胡程耀,黄培.固体溶解度测定方法的近期研究进展[J].药物分析杂志,2010,22(4):761-766.
[18]何钢,刘嵬,李会萍,等.银杏叶多糖分离纯化、结构鉴定及抗氧化活性研究[J].食品工业科技,2015,36(22):81-86.
[19]谢建华.青钱柳多糖的分子修饰及其生物活性研究[D].南昌:南昌大学,2014.
[20]夏朝红,戴奇,房韦,等.几种多糖的红外光谱研究[J].武汉理工大学学报,2007,29(1):45-47.
[21]周林珠,杨祥良,周井炎,等.多糖抗氧化作用研究进展[J].中国生化药物杂志,2002,23(4):210-212.
[22]Smirnoff N,Cumbes Q J.Hydroxyl radical scavenging activity of compatible solutes[J].Phytochemistry,1989,28(4):1057-1060.
[23]Yang S,Guo Z,Miao F,et al.The hydroxyl radical scavenging activity of chitosan,hyaluronan,starch and their O-carboxymethylated derivatives[J].Carbohydrate Polymers,2010,82(4):1043-1045.
[24]Xu Y,Niu X,Liu N,et al.Characterization,antioxidant and hypoglycemic activities of degraded polysaccharides from blackcurrant(Ribes nigrum,L.)fruits[J].Food Chemistry,2017:S0308814617315832.
[25]杨文鸽,谢果凰,徐大伦,等.龙须菜多糖的降解及其降解产物的抗氧化活性[J].水产学报,2009,33(2):342-347.
[26]宋见喜,任婷,王贺,等.高山红景天多糖的硫酸化修饰及DPPH自由基清除活性研究[J].中草药,2017,48(24):5125-5129.
[27]Kocharova N A,Thomas-Oates J E,Knirel Y A,et al.The structure of the O-specific polysaccharide of Citrobacter O16 containing glycerol phosphate[J].Febs Journal,2010,219(1-2):653-661.