蛋白酶在体外对豆粕球蛋白和β-伴球蛋白降解效果的研究
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摘要
本试验旨在体外评估几种蛋白酶对豆粕中球蛋白和β-伴球蛋白的降解效果,为饲用蛋白酶效果的快速评估提供依据。试验设4个处理(3个蛋白处理组和1个空白对照组),每个处理3个重复,蛋白酶DP100、蛋白酶J和蛋白酶K的添加量分别为1%、0.25%和0.3%,为各蛋白酶制剂产品推荐添加量的20倍。豆粕和豆粕加酶样品在相同的缓冲体系中酶解,用试剂盒检测豆粕酶解前后球蛋白和β-伴球蛋白的含量,进而评估不同蛋白酶对豆粕中球蛋白和β-伴球蛋白的降解效果。结果表明:豆粕中球蛋白和β-伴球蛋白的含量分别为13.47%和13.96%,占豆粕总蛋白含量的29.6%和30.3%;不同蛋白酶在体外对豆粕中球蛋白和β-伴球蛋白的降解幅度不同,蛋白酶DP100、蛋白酶J和蛋白酶K对豆粕中球蛋白降解率分别为73.3%、5%和5.9%,对豆粕中β-伴球蛋白的降解率分别52.1%、8.4%和0%。表明,蛋白酶DP100在体外对豆粕中球蛋白和β-伴球蛋白的降解效果明显优于蛋白酶J和蛋白酶K。
This study was conducted to investigate the hydrolysis ability of different proteases from China market on degrading glycinin and β-conglycinin of SBM in vitro. This in vitro trial included 4 treatments with 3 replicates per treatment. SBM used as the nature substrate was the control treatment, SBM supplemented with 3 different proteases were designed as the other 3 treatments. The adding dosage of Protease DP100, Protease J and Protease K collected from different suppliers was 1%, 0.25%, and 0.3% respectively, which was 20 times recommendation dosage of each supplier. The content of glycinin and β-conglycinin of SBM was measured by commercial testing kit before and after enzymatic hydrolysis. The results of this trial demonstrated that the hydrolysis rate of glycinin in SBM supplemented with DP100, Protease J and Protease K was 73.3%, 5% and 5.9% respectively, and the hydrolysis rate of β-conglycinin in SBM supplemented with DP100, Protease J and Protease K was 52.1%, 8.4% and 0% respectively. In conclusion, different protease products have different effects on allergic protein of SBM. Compared with Protease J and Protease K, DP100 protease was more efficient in degrading glycinin and β-conglycinin of SBM in vitro.
This study was conducted to investigate the hydrolysis ability of different proteases from China market on degrading glycinin and β-conglycinin of SBM in vitro. This in vitro trial included 4 treatments with 3 replicates per treatment. SBM used as the nature substrate was the control treatment, SBM supplemented with 3 different proteases were designed as the other 3 treatments. The adding dosage of Protease DP100, Protease J and Protease K collected from different suppliers was 1%, 0.25%, and 0.3% respectively, which was 20 times recommendation dosage of each supplier. The content of glycinin and β-conglycinin of SBM was measured by commercial testing kit before and after enzymatic hydrolysis. The results of this trial demonstrated that the hydrolysis rate of glycinin in SBM supplemented with DP100, Protease J and Protease K was 73.3%, 5% and 5.9% respectively, and the hydrolysis rate of β-conglycinin in SBM supplemented with DP100, Protease J and Protease K was 52.1%, 8.4% and 0% respectively. In conclusion, different protease products have different effects on allergic protein of SBM. Compared with Protease J and Protease K, DP100 protease was more efficient in degrading glycinin and β-conglycinin of SBM in vitro.
引文
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[12]周红蕾,李春玲,王贵平,等.大豆中抗营养因子及其去除方法概述[J].饲料工业,2006,27(3):23-26.
[13]Hei W J,Li Z,Ma X,et al.Determination of betaconglycinin in soybean and soybean products using a sandwich enzyme-linked immunosorbent assay[J].Anal Chim Acta,2012,734(1):62-68.
[14]邹婷婷,黑文静,贺平丽.双抗夹心ELISA检测β-伴大豆球蛋白方法的建立[A].中国畜牧兽医学会动物营养学分会第十一次全国动物营养学术研讨会论文集[C].北京:中国农业科学技术出版社,2012.
[2]Chen J,Wang J,Song P,et al.Determination of glycinin in soybean and soybean products using a sandwich enzymelinked immunosorbent assay[J].Food Chem,2014,(162):27-33.
[3]Hu S,Liu H,Qiao S,et al.Development of immunoaffinity chromatographic method for isolating glycinin(11S)from soybean proteins[J].J Agric Food Chem,2013,61(18):4406-4410.
[4]He L,Han M,Qiao S,et al.Soybean Antigen Proteins and their Intestinal Sensitization Activities[J].Curr Protein Pept Sci,2015,16(7):613-21.
[5]刘珊珊,武小霞,姜振峰,等.大豆7S球蛋白β-伴大豆球蛋白的研究现状[J].大豆科学,2007,26(3):417-422.
[6]杨玉娟,姚怡莎,秦玉昌,等.豆粕与发酵豆粕中主要抗营养因子调查分析[J].中国农业科学,2016,49(3):573-580.
[7]周天骄,谯仕彦,马曦,等.大豆饲料产品中主要抗营养因子含量的检测与分析[J].动物营养学报,2015,27(1):221-229.
[8]韩蕊.大豆球蛋白对仔猪小肠上皮细胞机械屏障功能的影响[D].长春:吉林农业大学,2013.
[9]Sun P,Li D F,Dong B,et al.Effects of soybean glycinin on performance and immune function in early weaned pigs[J].Arch Anim Nutr,2008,62(4):213-321.
[10]Chen F,Hao Y,Piao X S,et al.Soybean-derived betaconglycinin affects proteome expression in pig intestinal cells in vivo and in vitro[J].J Anim Sci,2010,89(3):743-753.
[11]Zhao Y,Qin G X,Sun Z W,et al.Effects of glycinin andβ-conglycinin on enterocyte apoptosis,proliferation and migration of piglets[J].Food Agr Immunol,2010,21(3):209-218.
[12]周红蕾,李春玲,王贵平,等.大豆中抗营养因子及其去除方法概述[J].饲料工业,2006,27(3):23-26.
[13]Hei W J,Li Z,Ma X,et al.Determination of betaconglycinin in soybean and soybean products using a sandwich enzyme-linked immunosorbent assay[J].Anal Chim Acta,2012,734(1):62-68.
[14]邹婷婷,黑文静,贺平丽.双抗夹心ELISA检测β-伴大豆球蛋白方法的建立[A].中国畜牧兽医学会动物营养学分会第十一次全国动物营养学术研讨会论文集[C].北京:中国农业科学技术出版社,2012.