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| Secretory Expression of Nattokinase from Bacillus natto in Escherichia coli |
| HAN Hui-ming1, LI Yong-mei2 |
1. Basic Medical College, Beihua University, Jilin 132013, China;
2. School of Stomatology, Beihua University, Jilin 132013, China |
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Abstract Nattokinase has been reported as an oral health product for the prevention of atherosclerosis. A novel strategy to express a nattokinase from Bacillus natto in a live delivery vehicle E. coli BL21(DE3) was developed. The mature peptide-coding fragment of NK gene was amplified from B. natto total DNA by polymerase chain reaction (PCR). The sequence analysis indicated that the fragment of NK gene consisted of 825 bp, which encoded 275 amino acid residues. The sequence of NK gene has 99% homology with those of the reported ones. The expression plasmids NK-pep-pMAL-p2X and NK-pep-pMAL-c2X are constructed by inserting NK gene into vector pMAL-p2X and pMAL-c2X, and then fusionally expressed with maltose-binding protein (MBP) in Escherichia coli BL21(DE3). SDS-PAGE analysis revealed that the NK protein were secretion expressed after induction with IPTG for 4 hours under 16℃ or 37℃.The characteristic of this recombinant nattokinase is comparable to the native nattokinase. Secretory expression of nattokinase in E. coli would facilitate the development of this enzyme into a therapeutic product for the control and prevention of thrombosis diseases.
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Received: 12 August 2014
Published: 25 October 2014
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