| 亲和层析法分离纯化纳豆激酶 |
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| 引用本文： | 刘柳,李南薇,郭勇.亲和层析法分离纯化纳豆激酶[J].食品科学,2011,32(16):58-61. |
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| 作者姓名： | 刘柳 李南薇 郭勇 |
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| 作者单位： | 1.暨南大学食品科学与工程系 2.仲恺农业工程学院轻工食品学院 3.华南理工大学生物科学与工程学院 |
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| 摘 要： | 目的：以纳豆激酶的作用底物纤维蛋白为配基制备亲和层析胶，分离纯化纳豆激酶。方法：纳豆杆菌发酵液经硫酸铵分段盐析、透析得粗酶，在4℃条件亲和层析法分离纯化，利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate polyacrylamide gel electrophoresis，SDS-PAGE)对酶纯度进行检验。结果：经亲和层析得纳豆激酶为电泳纯，纯化倍数为8.3，回收率43.9%。纯酶的最适pH值和温度分别为pH7.0～9.0、50℃；温度高于60℃纤溶酶活性迅速下降；在pH6.0～10.0范围内稳定性好；Mg2＋对其有微弱激活作用，Cu2+有显著抑制作用。结论：以琼脂糖包埋纤维蛋白制备的层析胶可用于纳豆激酶的快速分离纯化。
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| 关 键 词： | 纳豆激酶 分离纯化 亲和层析 纤维蛋白 酶学性质 |
| 收稿时间： | 2018-05-21 |
Affinity Chromatographic Purification of Nattokinase |
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| LIU Liu,LI Nan-wei,GUO Yong.Affinity Chromatographic Purification of Nattokinase[J].Food Science,2011,32(16):58-61. |
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| Authors: | LIU Liu LI Nan-wei GUO Yong |
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| Affiliation: | (1. Department of Food Science and Engineering, Jinan University, Guangzhou 510632, China；
2. College of Light Industry and Food Technology, Zhongkai University of Agricultural and Engineering, Guangzhou 510225, China；
3. School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510641, China) |
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| Abstract: | Objective: To separate and purify nattokinase by fibrin affinity chromatography. Methods: The fermentation supernatant of Bacillus subtilis subsp. natto was fractionally salted out with ammonium sulfate and dialyzed, and the crude nattokinase obtained was purified by affinity chromatographic column chromatography at 4 ℃. The purity of purified nattokinase was analyzed by SDS-PAGE. Results: Nattokinase of electrophoretical purity was obtained, with a purify fold of 8.3 and a recovery of 43.9%. The optimum reaction pH and temperature pf purified nattokinase were 7.0－9.0 and 50 ℃, respectively. The enzyme activity exhibited a sharp drop at temperatures above 60 ℃ and good stability in the pH range of 6.0－10.0. The enzyme was slightly activated by Mg2+ but dramatically inhibited by Cu2+. Conclusion: Affinity chromatography on fibrin-agarose is applicable to fast purify nattokinase. |
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| Keywords: | nattokinase purification affinity chromatography fibrin characterization |
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