基于分光光度法构建纳豆激酶纤溶活性快速测定方法

聂光军; 赵锐; 岳文瑾; 杨超英; 薛正莲; 魏明; 温东升; 聂智杰; 叶裕超

doi:10.13386/j.issn1002-0306.2015.13.054

基于分光光度法构建纳豆激酶纤溶活性快速测定方法

安徽工程大学,生物与化学工程学院
安徽工程大学,物与化学工程学院

基金项目:

安徽省自然科学基金(1508085SMC212)；安徽省高校自然科学研究重点项目(KJ2014A025)；国家级大学生创新创业训练计划(201210363042,201310363076)；

详细信息

作者简介:
聂光军 (1976-) , 男, 博士, 副教授, 研究方向:分子生物学。;

岳文瑾 (1979-) , 女, 博士, 副教授, 研究方向:生物纳米材料。;

中图分类号: O657.3;TQ464.8
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出版历程
- 收稿日期: 2014-07-27

Construction of a rapid method for assaying the fibrinolytic activity of Nattokinase based on spectrophotometric assay method

College of Biochemical Engineering, Anhui Polytechnic University

摘要

摘要: 为建立一种快捷、有效的纳豆激酶纤溶活性测定方法,在研究纤维蛋白平板法的基础上,构建了以透明圈面积为中间参数的纳豆激酶纤溶活力与吸光度值之间关系模型,通过测定纳豆激酶显色反应中的吸光度值,直接获得纳豆激酶的纤溶活性。结果显示改进后的酪蛋白方法可行,其模型方程为y=1877.77x-481.08（R2=0.9924）。验证结果显示模型相关性强,准确度较高,操作简便、快速、检测成本低,是一种高效的纳豆激酶纤溶活性快速检测方法。
- 纳豆激酶 /
- 纤维蛋白平板法 /
- 酪蛋白法 /
- TAME法
Abstract: To construct a rapid, efficient method for assaying the fibrinolytic activity of nattokinase, two models between fibrinolytic activity of nattokinase in fibrin plate method and absorbance value of nattokinase in casein method or in TAME method were formed with the size of transparent zone as an intermediate parameter based on the study on fibrin plate method. The fibrinolytic activity can be directly determined by detecting the absorbance value of nattokinase in casein method or in TAME method.The results showed that the improved casein method can be applied in determining the fibrinolytic activity of nattokinase, the model in the improved method was as the following: y = 1877.77x-481.08 ( R2= 0.9924) .The result of verification experiment indicated that the model owned such advantages as strong correlation, high degree of accuracy, easy operation, high efficiency, and low cost.Therefore, the improved casein method was an efficient method for real time detecting the fibrinolytic activity of nattokinase.
- Nattokinase /
- fibrin plate method /
- casein method /
- TAME method

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参考文献(14)

[1]	Chen R P, Lee K.Nattokinase for degrading and reducing amyloid fibrils-associated with alzheimer’s disease, prion diseases and other amyloidoses[P].US Patents:US8137666B2;2012.
[2]	Wu S M, Feng C, Zhong J, et al.Enhanced production of recombinant nattokinase in Bacillus subtilis by promoter optimization[J].World Journal of Microbiology and Biotechnology, 2011, 27 (1) :99-106.
[3]	M M, M K.Natto and its acrive ingredient nattokinase a potent and safe thrombolytic agent[J].Alternative and Complementary Therapies, 2002, 8 (3) :157-164.
[4]	纪红蕊.纳豆激酶最新研究进展[J].亚太传统医药, 2010, 6 (5) :144-146.
[5]	邹和昌.溶栓剂的发展及研究[J].中国药学杂志, 1997, 32 (5) :263.
[6]	Liu C, Wu W, Lai H.Adsorption of Nattokinase by Amino Acid-Conjugated Magnetic Nanoadsorbents[J].Separation Science and Technology, 2013, 48 (6) :923-930.
[7]	杨明俊, 杨晓彤, 冯慧琴, 等.两种纳豆激酶活性测定方法对比及相关性分析[J].食品研究与开发, 2008, 29 (2) :137-141.
[8]	Sumi H, Hamada H, Tsushima H, et al.A novel fibrinolytic enzyme (nattokinase) in the vegetable cheese Natto;a typical and popular soybean food in the Japanese diet[J].Experientia, 1987, 43 (10) :1110-1111.
[9]	Hara T, Tadokoro Y, Satoyama T.A simple, easy and routine assay of fibrinolytic enzyme activity[J].Journal of Japanese Society for Food Science and Technology, 1996, 43 (2) :172-175.
[10]	秋玲, 林剑, 张玲.溶血栓583酶活性测定的一种新方法[J].生物化学与生物物理进展, 2001, 28 (4) :582-583.
[11]	魏华, 赵祥颖, 刘建军.纳豆激酶的活性测定[J].山东轻工业学院学报, 2007, 21 (1) :60-63.
[12]	马恩平, 陈超, 夏泉, 等.纳豆激酶活性的检测及纳豆粉筛选[J].食品科学, 2012, 37 (10) :67-70.
[13]	熊迎新, 尹宗宁, 杨超, 等.纳豆激酶活性测定方法的研究[J].药物生物技术, 2006, 13 (2) :140-143.
[14]	杨明, 董超, 史延茂, 等.纤维蛋白平板法测定纳豆激酶方法的改进[J].中国酿造, 2008, 27 (7) :77-80.