| Prokaryotic expression, purification and characterization of nattokinase |
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| 作者姓名： | AI Hai-xin ZHANG Li ZHANG Xin-gang CHEN Si-yao HU Huan WEI Hong-yun MENG Xin-rui WANG Tian-qi ZHAO Jian LIU Hong-sheng |
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| 作者单位： | 1.School of Life Science,Liaoning University, Shenyang 110036, China;2.Research Center for Computer Simulating andInformation Processing of Bio-macromolecules of Liaoning, Shenyang 110036, China;3.Liaoning Engineering Laboratory for Molecular Simulation and Designing of Drug Molecules, Shenyang 110036, PR China;,1.School of Life Science,Liaoning University, Shenyang 110036, China;2.Research Center for Computer Simulating andInformation Processing of Bio-macromolecules of Liaoning, Shenyang 110036, China,School of Life Science,Liaoning University, Shenyang 110036, China,School of Life Science,Liaoning University, Shenyang 110036, China,1.School of Life Science,Liaoning University, Shenyang 110036, China;3.Liaoning Engineering Laboratory for Molecular Simulation and Designing of Drug Molecules, Shenyang 110036, PR China;,1.School of Life Science,Liaoning University, Shenyang 110036, China;4.Sion Tiansheng Biological Technology Co.,Ltd.,Shenyang 110036,PR.China,School of Life Science,Liaoning University, Shenyang 110036, China,School of Pharmacy, Liaoning University, Shenyang 110036,School of Life Science,Liaoning University, Shenyang 110036, China,1.School of Life Science,Liaoning University, Shenyang 110036, China;2.Research Center for Computer Simulating andInformation Processing of Bio-macromolecules of Liaoning, Shenyang 110036, China;3.Liaoning Engineering Laboratory for Molecular Simulation and Designing of Drug Molecules, Shenyang 110036, PR China; |
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| 基金项目： | 辽宁省科技厅基金资助项目（2013225086,2014001015）；辽宁省教育厅基金资助项目（L2014001,LT2015011）；沈阳市科技局基金资助项目（F15165400,F16205151） |
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| 摘 要： | Nattokinase is a fibrinolytic enzyme that is considered to be a promising agent for thrombosis therapy. In this study, nattokinase was purified from fermentation broth of a Bacillus subtilis strain by ammonium sulfate salting-out, gel filtration chromatography, and hydrophobic interaction chromatography with a purification fold of 5.2 and at a yield of 46.3%. The purified enzyme has molecular mass of 28 kDa and fibrinolytic activity of 4 580 U/mg. Since the concentration of nattokinase on fermentation broth was quite low, we cloned nattokinase gene from B. subtilis and expressed it in E. coli BL21 (DE3). Nattokinase was actively expressed in the recombinant strain. The yield of nattokinase was increased significantly, but the activity of the protein produced by recombinant strain was low.
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| 关 键 词： | nattokinase purification prokaryotic expression |
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