北冬虫夏草多肽分离纯化及活性研究
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摘要
目的对北冬虫夏草活性多肽进行分离纯化,并测定其抗氧化活性.方法优选北冬虫夏草多肽的提取方法;通过体外抗氧化DPPH清除率测定北冬虫夏草多肽的抗氧化活性;通过大孔树脂HP-20对北冬虫夏草多肽产物进行分离纯化,并分别测定洗脱后各浓度产物的蛋白含量和抗氧化活性,经Tricine-SDS-PAGE电泳法初步鉴定北冬虫夏草中多肽的分子量.结果 Tris-HCl作为提取液时DPPH清除率最高可达88.68%;经大孔树脂HP-20洗脱并分离纯化后,25%浓度的乙醇溶液和去离子水可作为洗脱剂; Tricine-SDS-PAGE结果显示:北冬虫夏草中富含小分子肽.结论此研究确定了北冬虫夏草多肽的最佳提取工艺,提高了初步提取多肽的产量;北冬虫夏草中的多肽具有一定的抗氧化活性.
Objective The active peptide of Cordyceps militaris was isolated and purified,and its antioxidant activity was determined. Method The extraction method of Cordyceps militaris polypeptide was optimized. The antioxidant activity of Cordyceps militaris polypeptide was determined by in vitro anti-oxidation DPPH clearance rate.The macroporous resin HP-20 was used to separate and purify the polypeptide of Cordyceps militaris,and the products after elution were determined. Protein content and antioxidant activity,the molecular weight of the polypeptide in Cordyceps militaris were identified by Tricine-SDS-PAGE electrophoresis. Results The DPPH scavenging rate of Tris-HCl as extract is up to 88.68%.After elution and purification by macroporous resin HP-20,25% ethanol solution and deionized water can be used as eluent; Tricine-SDS-PAGE results showed that Cordyceps militaris was rich in small peptides.Conclusion The optimal extraction process of Cordyceps militaris polypeptide was determined,and the yield of the preliminary extracted peptide was improved. The polypeptide of Cordyceps militaris had certain antioxidant activity.
Objective The active peptide of Cordyceps militaris was isolated and purified,and its antioxidant activity was determined. Method The extraction method of Cordyceps militaris polypeptide was optimized. The antioxidant activity of Cordyceps militaris polypeptide was determined by in vitro anti-oxidation DPPH clearance rate.The macroporous resin HP-20 was used to separate and purify the polypeptide of Cordyceps militaris,and the products after elution were determined. Protein content and antioxidant activity,the molecular weight of the polypeptide in Cordyceps militaris were identified by Tricine-SDS-PAGE electrophoresis. Results The DPPH scavenging rate of Tris-HCl as extract is up to 88.68%.After elution and purification by macroporous resin HP-20,25% ethanol solution and deionized water can be used as eluent; Tricine-SDS-PAGE results showed that Cordyceps militaris was rich in small peptides.Conclusion The optimal extraction process of Cordyceps militaris polypeptide was determined,and the yield of the preliminary extracted peptide was improved. The polypeptide of Cordyceps militaris had certain antioxidant activity.
引文
[1]范克明.蛹虫草诱变菌株多糖的提取及其抗氧化活性分析[D].济南:山东农业大学,2011.
[2]漆伟,雷伟,严亚波,等.冬虫夏草药理学作用的研究进展[J].环球中医药,2014,7(3):227-232.
[3]张杰,李春艳,李劲平,等.蒽酮硫酸法与苯酚硫酸法测定竹节参多糖含量的比较研究[J].中南药学,2012,10(6):421-424.
[4]李桂梅.PDCA循环在医院药学管理中的应用[J].中国保健营养,2013,4(1):301-303.
[5] Li S P,Zhang G H,Zeng Q,et al.Hypoglycemic activity of polysaccharide,with antioxidation,isolated from cultured Cordycepsmycelia[J].Phytomedicine,2006,13(6):423-428.
[6]元玲刚,常明昌,刘靖宇,等.北虫草多糖提取工艺优化及抗氧化作用研究[J].中国农学通报,2015,382(19):144-148.
[7]刘春泉,宋江峰,李大婧,等.北冬虫夏草多肽组分的分离纯化及结构研究进展[J].食品科学,2007,28(1):370-373.
[8]邵颖,袁凤健,陈安徽,等.富硒蛹虫草活性成分及抗氧化作用分析[J].食品工业,2013,34(4):126-129.
[9]翁梁,温鲁,杨芳,等.不同提取方法对蛹虫草多糖抗氧化性的影响[J].食品科技,2008,33(11):180-182.
[10] Bermhard E J.Inhibiting Rasprenylation increases the radio sensitivity of human tumor cell lines with activ-ating mutations ofrasoncogenes[J].Cancer Res,1998,58(8):1754-1761.
[11]赵秋蓉,李建平,吴迪,等.冬虫夏草中多糖提取、纯化及抗氧化性能的研究[J].中国农学通报,2012,28(15):238-242.
[12]张岚.干巴菌多糖的分离提取、纯化及其抗氧化活性研究[D].昆明:云南农业大学,2017.
[13] Shuhan Chen,Haixia Chen,Zhaoshuai Wang,et al. Structural characterization and antioxidant properties of polysaccharidesfrom the fruiting bodies of Russula virescens[J].Food Chemistry,2010,118:675-680.
[14] Siqian Li,Nagendra P Shah.Antioxidant and antiba-cterial activities of sulphated polysaccharides from Pleurotus eryngii andStreptococcus thermophilus[J].Food Chemistry,2014,165:262-270.
[15] Debnath T,Park P J,Nath NCD,et al.Antioxidant activity of Gardenia jasminoides Ellis fruit extracts[J].Food Chemistry,2011,128:697-703.
[2]漆伟,雷伟,严亚波,等.冬虫夏草药理学作用的研究进展[J].环球中医药,2014,7(3):227-232.
[3]张杰,李春艳,李劲平,等.蒽酮硫酸法与苯酚硫酸法测定竹节参多糖含量的比较研究[J].中南药学,2012,10(6):421-424.
[4]李桂梅.PDCA循环在医院药学管理中的应用[J].中国保健营养,2013,4(1):301-303.
[5] Li S P,Zhang G H,Zeng Q,et al.Hypoglycemic activity of polysaccharide,with antioxidation,isolated from cultured Cordycepsmycelia[J].Phytomedicine,2006,13(6):423-428.
[6]元玲刚,常明昌,刘靖宇,等.北虫草多糖提取工艺优化及抗氧化作用研究[J].中国农学通报,2015,382(19):144-148.
[7]刘春泉,宋江峰,李大婧,等.北冬虫夏草多肽组分的分离纯化及结构研究进展[J].食品科学,2007,28(1):370-373.
[8]邵颖,袁凤健,陈安徽,等.富硒蛹虫草活性成分及抗氧化作用分析[J].食品工业,2013,34(4):126-129.
[9]翁梁,温鲁,杨芳,等.不同提取方法对蛹虫草多糖抗氧化性的影响[J].食品科技,2008,33(11):180-182.
[10] Bermhard E J.Inhibiting Rasprenylation increases the radio sensitivity of human tumor cell lines with activ-ating mutations ofrasoncogenes[J].Cancer Res,1998,58(8):1754-1761.
[11]赵秋蓉,李建平,吴迪,等.冬虫夏草中多糖提取、纯化及抗氧化性能的研究[J].中国农学通报,2012,28(15):238-242.
[12]张岚.干巴菌多糖的分离提取、纯化及其抗氧化活性研究[D].昆明:云南农业大学,2017.
[13] Shuhan Chen,Haixia Chen,Zhaoshuai Wang,et al. Structural characterization and antioxidant properties of polysaccharidesfrom the fruiting bodies of Russula virescens[J].Food Chemistry,2010,118:675-680.
[14] Siqian Li,Nagendra P Shah.Antioxidant and antiba-cterial activities of sulphated polysaccharides from Pleurotus eryngii andStreptococcus thermophilus[J].Food Chemistry,2014,165:262-270.
[15] Debnath T,Park P J,Nath NCD,et al.Antioxidant activity of Gardenia jasminoides Ellis fruit extracts[J].Food Chemistry,2011,128:697-703.