海藻酸寡糖的纯化及分子量测定
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摘要
通过酶解法制备出海藻酸寡糖粗品,采用Q Sepharose FF离子交换柱层析对寡糖进行分离纯化获得AS1和AS2,分别用凝胶层析法和乌式黏度法测定其相对分子质量。结果表明,凝胶层析法更适合寡糖分子量的测定,SA1和SA2的相对分子量为别为1.0×10~4Da、5.8×10~3Da。为海藻酸寡糖的进一步研究及开发提供了科学依据。
Alginic acid oligosaccharide AS1 and AS2 were degraded by enzymatic hydrolysis,using Q Sepharose FF anion-exchange column chromatography of oligosaccharide separation,and their molecular weight were determined by gel chromatography method and ubbelohde viscometer. Results showed that two alginic acid oligosaccharide components of AS1 and AS2 were more suitable for the determination through gel chromatography method. The relative molecular weight of AS1 and AS2 components were 1.0×10~4 Da、5.8×10~3 Da,respectively. The experiment can be used as a scienti?c basis for the research of alginic acid oligosaccharide in the future.
Alginic acid oligosaccharide AS1 and AS2 were degraded by enzymatic hydrolysis,using Q Sepharose FF anion-exchange column chromatography of oligosaccharide separation,and their molecular weight were determined by gel chromatography method and ubbelohde viscometer. Results showed that two alginic acid oligosaccharide components of AS1 and AS2 were more suitable for the determination through gel chromatography method. The relative molecular weight of AS1 and AS2 components were 1.0×10~4 Da、5.8×10~3 Da,respectively. The experiment can be used as a scienti?c basis for the research of alginic acid oligosaccharide in the future.
引文
[1]欧昌荣.褐藻胶寡糖的制备、分析及其应用研究[D].中国海洋大学,2003.
[2]包华芳,刘璘,丁玉庭.酶解制备褐藻胶寡糖及其抗氧化活性研究[J].中国酿造,2010(4):82-84.
[3]Thiang Y W,Lori A P,Neal L S.Alginate lyase:Review of major sources and enzyme characteristic,structure-function analysis,biological roles,and applications[J].Annual Review of Microbiology,2000,54(2):289-340.
[4]王媛媛,郭文斌,王淑芳,等.褐藻寡糖的生物活性与应用研究进展[J].食品与发酵工业,2010,36(10):122-126.
[5]Hu X,Jiang X.Antitumour activities of alginate-derived oligosaccharides and their sulphated substitution derivatives[J].European Journal of Phycology,2004,39(1):67-71.
[6]刘瑞志.褐藻寡糖促进植物生长与抗逆效应机理研究[D].中国海洋大学,2009.
[7]张真庆.褐藻胶酶解寡糖的制备、分离和结构鉴定[D].中国海洋大学,2003.
[8]赵珊,许加超,付晓婷,等.褐藻寡糖分子量测定方法的研究[J].食品工业科技,2011,32(12):486-488.
[9]Uwe Remminghorst,Bernd H A Rehm.Bacterial alginates:from biosynthesis to applications[J].Biotechnol Lett,2006,28:1701-1712.
[10]Hirsh J,Anand SS,Halperin J,et al.Guide to anticoagulant therapy:heparin.A statement for healthcare professionals from the American Heart Association[J].Circulation,2001,103(24):2994-3018.
[11]张明杰,李恒,蒋敏,等.低分子质量褐藻寡糖的高效酶法制备及其抗氧化活性评价[J].海洋科学,2016,40(12):62-70.
[12]陈丽,张林维,薛婉立.褐藻寡糖的制备及抑菌性研究[J].中国饲料,2007(9):34-36.
[13]甘纯玑,林权.褐藻酸钠的高效凝胶色谱行为及其分子量参数测定[J].色谱,1997,15(2):147-149.
[14]甘纯玑.褐藻酸钠溶液的超声辐照效应及其对分子量参数的影响[J].高等学校化学学报,1995,16(12):1956-1959.
[15]李潇.褐藻酸寡糖的分离分析及其抗氧化活性的研究[D].西北大学,2011.
[16]吴扬兰,王远亮,杨亚联,等.GPC-十八角激光光散射联用法测定聚乳酸的分子量及分子量分布[J].现代科学仪器,1997,2(8):87-90.
[17]成跃祖.凝胶渗透色谱法的进展及其应用[J].中国石化出版社,1993,54(11):5-14.
[18]李林,罗琼,张声华.海带多糖的分类提取、鉴定及理化特性研究[J].食品科学,2000,21(4):28-32.第二十四届中国国际食暨第三十届全国食品添加Food Ingredien展出时间:2020年展出地点:国家会展中心详情请登陆:展出时间:2020年展出地点:国家会展中心详情请登陆:
[19]张惟杰.糖复合物生化研究技术第二版[M].浙江大学出版社,1999,133.
[20]付昊雨,张大伟,吴庆燕,等.红松松子壳多糖分离纯化及对小鼠抗氧化活性的影响[J].吉林农业大学学报,2017(2):1-4.
[21]徐光域,颜军,郭晓强,等.硫酸-苯酚定糖法的改进与初步应用[J].食品科学,2005,26(8):342-346.
[22]冯孟鑫.人参果多糖的分级分离纯化及理化性质与抗肿瘤活性初探[D].北京中医药大学,2016.
[23]Song C F,Wang S G,Yang J,et al.Optimization of Vacuum-Microwave Radiation Pretreatment on Extraction of Ganoderma Polysaccharides[J].Mathematical Problems in Engineering,2015(4):113-116.
[24]聂少平,谢明勇,申明月,等.应用高效液相色谱法测定茶叶多糖[J].食品科学,2006,27(4):177-181.
[25]Olav smidsrqd and Arne haug.A light scattering study of alginate[J].Acta Chemical Scandinavica,1968,22:797-810.
[26]安晓娟,冯琳,宋红平,等.植物多糖的结构分析及药理活研究进展[J].中国药学杂志,2012,47(16):1271-1275.
[27]马永强,张静,王鑫,等.酶法提取甜玉米芯多糖的对比研究[J].哈尔滨商业大学学报,2015(4):162-165.品添加剂和配料展览会加剂生产应用技术展示会nts China 2020nts China 2020年3月17日-19日(上海),崧泽大道333号:www.cfaa.cn年3月17日-19日(上海),崧泽大道333号:www.cfaa.cn
[2]包华芳,刘璘,丁玉庭.酶解制备褐藻胶寡糖及其抗氧化活性研究[J].中国酿造,2010(4):82-84.
[3]Thiang Y W,Lori A P,Neal L S.Alginate lyase:Review of major sources and enzyme characteristic,structure-function analysis,biological roles,and applications[J].Annual Review of Microbiology,2000,54(2):289-340.
[4]王媛媛,郭文斌,王淑芳,等.褐藻寡糖的生物活性与应用研究进展[J].食品与发酵工业,2010,36(10):122-126.
[5]Hu X,Jiang X.Antitumour activities of alginate-derived oligosaccharides and their sulphated substitution derivatives[J].European Journal of Phycology,2004,39(1):67-71.
[6]刘瑞志.褐藻寡糖促进植物生长与抗逆效应机理研究[D].中国海洋大学,2009.
[7]张真庆.褐藻胶酶解寡糖的制备、分离和结构鉴定[D].中国海洋大学,2003.
[8]赵珊,许加超,付晓婷,等.褐藻寡糖分子量测定方法的研究[J].食品工业科技,2011,32(12):486-488.
[9]Uwe Remminghorst,Bernd H A Rehm.Bacterial alginates:from biosynthesis to applications[J].Biotechnol Lett,2006,28:1701-1712.
[10]Hirsh J,Anand SS,Halperin J,et al.Guide to anticoagulant therapy:heparin.A statement for healthcare professionals from the American Heart Association[J].Circulation,2001,103(24):2994-3018.
[11]张明杰,李恒,蒋敏,等.低分子质量褐藻寡糖的高效酶法制备及其抗氧化活性评价[J].海洋科学,2016,40(12):62-70.
[12]陈丽,张林维,薛婉立.褐藻寡糖的制备及抑菌性研究[J].中国饲料,2007(9):34-36.
[13]甘纯玑,林权.褐藻酸钠的高效凝胶色谱行为及其分子量参数测定[J].色谱,1997,15(2):147-149.
[14]甘纯玑.褐藻酸钠溶液的超声辐照效应及其对分子量参数的影响[J].高等学校化学学报,1995,16(12):1956-1959.
[15]李潇.褐藻酸寡糖的分离分析及其抗氧化活性的研究[D].西北大学,2011.
[16]吴扬兰,王远亮,杨亚联,等.GPC-十八角激光光散射联用法测定聚乳酸的分子量及分子量分布[J].现代科学仪器,1997,2(8):87-90.
[17]成跃祖.凝胶渗透色谱法的进展及其应用[J].中国石化出版社,1993,54(11):5-14.
[18]李林,罗琼,张声华.海带多糖的分类提取、鉴定及理化特性研究[J].食品科学,2000,21(4):28-32.第二十四届中国国际食暨第三十届全国食品添加Food Ingredien展出时间:2020年展出地点:国家会展中心详情请登陆:展出时间:2020年展出地点:国家会展中心详情请登陆:
[19]张惟杰.糖复合物生化研究技术第二版[M].浙江大学出版社,1999,133.
[20]付昊雨,张大伟,吴庆燕,等.红松松子壳多糖分离纯化及对小鼠抗氧化活性的影响[J].吉林农业大学学报,2017(2):1-4.
[21]徐光域,颜军,郭晓强,等.硫酸-苯酚定糖法的改进与初步应用[J].食品科学,2005,26(8):342-346.
[22]冯孟鑫.人参果多糖的分级分离纯化及理化性质与抗肿瘤活性初探[D].北京中医药大学,2016.
[23]Song C F,Wang S G,Yang J,et al.Optimization of Vacuum-Microwave Radiation Pretreatment on Extraction of Ganoderma Polysaccharides[J].Mathematical Problems in Engineering,2015(4):113-116.
[24]聂少平,谢明勇,申明月,等.应用高效液相色谱法测定茶叶多糖[J].食品科学,2006,27(4):177-181.
[25]Olav smidsrqd and Arne haug.A light scattering study of alginate[J].Acta Chemical Scandinavica,1968,22:797-810.
[26]安晓娟,冯琳,宋红平,等.植物多糖的结构分析及药理活研究进展[J].中国药学杂志,2012,47(16):1271-1275.
[27]马永强,张静,王鑫,等.酶法提取甜玉米芯多糖的对比研究[J].哈尔滨商业大学学报,2015(4):162-165.品添加剂和配料展览会加剂生产应用技术展示会nts China 2020nts China 2020年3月17日-19日(上海),崧泽大道333号:www.cfaa.cn年3月17日-19日(上海),崧泽大道333号:www.cfaa.cn