大豆凝集素纯化及凝集活性测定的研究
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摘要
通过建立快速大豆凝集素(SBA)粗提液制备方法,利用N-乙酰-D-半乳糖胺-epoxy-Sepharose 6B亲和层析制备体系制备大豆凝集素样品,采用聚丙烯酰胺(SDS-PAGE)及红细胞凝集试验分别对本试验制备的大豆凝集素进行纯度及凝集活性的检测。结果表明,试验制备的大豆凝集素的产率为135 mg/100 g;所制得的大豆凝集素样品的纯度较高,与大豆凝集素标准品基本一致;其凝集活性为256。研究成果可为大豆凝集素的量化生产提供技术参考。
引文
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[2]Bajpai S,Sharma A,Gupta M N.Removal and recovery of antinutritional factors from soybean flour[J].Food Chemistry,2005,89(4):497-501.
[3]Fasina Y O,Classen H L,Garlich J D,et al.Investigating the possibility of monitoring lectin levels in commercial soybean meals intended for poultry feeding using steam-heated soybean meal as a model[J].Poultry Science,2003,82(4):648-656.
[4]张柏林,秦贵信,刘宁,等.大豆凝集素结构及其活性测定方法的研究进展[J].大豆科学,2009,28(1):160-163.
[5]王利民.大豆凝集素在不同种属动物体内的消化动力学及抗营养作用比较研究[D].长春:吉林农业大学,2007.
[6]Franco-Fraguas L,Alicia P,Ferreira F,et al.Preparative purification of soybean agglutinin by affinity chromatography and its immobilization for polysaccharide isolation[J].Journal of Chromatography B,2003,790(1/2):365-372.
[7]Liener E I.Implications of antinutritional components in soybean foods[J].Critical Reviews in Food Science and Nutrition,1994,34(1):31-67.
[8]Liener E I,Pallansch M J,et al.Purification of a toxic substance from defatted soy bean flour[J].Journal of Biological Chemistry,1952,197(1):29-36.
[9]Wada S,Pallansch M J,Liener I E,et al.Chemical composition and end groups of the soybean hemagglutinin[J].Journal of Biological Chemistry,1958,233(2):395-400.
[10]张洪渊,刘克武,杨守忠,等.大豆凝集素、脲酶和胰蛋白酶抑制剂的分离纯化[J].中国油料作物学报,1991(1):41-45.
[11]Vretblad P.Purification of lectins by biospecific affinity chromatography[J].BBA-Protein structure,1976,434(1):169-176.
[12]Li Z T,Li D,Qiao S Y,et al.Anti-nutritional effects of a moderate dose of soybean agglutinin in the rat[J].Archiv Fur Tierernahrung,2003,57(4):267-277.
[13]王利民,秦贵信,刘林娜,等.三种亲和层析体系纯化大豆凝集素的比较研究[J].大豆科学,2007,26(1):51-54.
[14]李振田.大豆凝集素的检测、纯化和对大鼠抗营养机理的研究[D].北京:中国农业大学,2003.
[15]胡海霞.大豆凝集素与不同种属动物肠壁结合规律的比较研究[D].长春:吉林农业大学,2008.
[16]何昭阳.动物免疫学实验技术[M].长春:吉林科学技术出版社,2002.
[17]郭尧君.蛋白质电泳实验技术[M].北京:科学出版社,1999.
[18]Lotan R,Siegelman H W,Lis H,et al.Subunit structure of soybean agglutinin[J].Journal of Biological Chemistry,1974,249(4):1219-1224.
[19]Jaffe G W.In toxic constituents in plant foodstuffs[M].New York:Academic Press,1980.
[20]刘林娜.不同种属动物对大豆凝集素敏感性的比较研究[D].长春:吉林农业大学,2006.
[21]Pusztai A,Watt W B,Stewart J C,et al.A comprehensive scheme for the isolation of trypsin inhibitors and the agglutinin from soybean seeds[J].Journal of Agricultural and Food Chemistry,2002,39(5):862-866.
[22]Liener I E.Comment on a semi-pilot-scale procedure for isolating and purifying soybean(Glycine max)lectin[J].Journal of Agricultural and Food Chemistry,2003,51(16):4532-4538.