Extracellular Production of a Potent and Chemical Resistant Nattokinase in Immobilized Escherichia coli Using Response Surface Methodology.

Extracellular Production of a Potent and Chemical Resistant Nattokinase in Immobilized Escherichia coli Using Response Surface Methodology. Curr Pharm Biotechnol. 2018 Oct 22;: Authors: Mohammadi F, Nezafat N, Berenjian A, Negahdaripour M, Zamani M, Ghoshoon MB, Morowvat MH, Hemmati S, Ghasemi Y Abstract Nattokinase is a potent fibrinolytic protease, besides various industrial applications used as a drug for treatment or a supplement for preventing thrombosis. Generally, most of native extracellular proteins such as nattokinase from B.subtilis are lysed by some secretory proteases. Therefore, one way for solving this problem, is to apply other hosts for nattokinase production. There are different factors for producing secretory form of nattokinase from B.subtilis, such as suitable host, optimized media for the maximum activity and easy purification which are studied in this investigation. E.coli BL21 (DE3), as a reliable host was selected for production of a high yield of an ectracellular recombinant nattokinase. A mature nattokinase gene from Bacillus subtilis 1023, was cloned in the expression vector pET22b and transformed in the host. The recombinant nattokinase was expressed by 1mM IPTG induction for 4 hours. The expressed protein was confirmed by SDS-PAGE, and its fibrinolytic activity was assayed on the fibrin plates. Afterwards, the enzyme was purified by Ni-NTA native affinity column. Evaluating different media components for...
Source: Current Pharmaceutical Biotechnology - Category: Biotechnology Authors: Tags: Curr Pharm Biotechnol Source Type: research