Extracellular Production of a Potent and Chemical Resistant Nattokinase in Immobilized Escherichia coli Using Response Surface Methodology.
Authors of this article are:
Mohammadi F, Nezafat N, Berenjian A, Negahdaripour M, Zamani M, Ghoshoon MB, Morowvat MH, Hemmati S, Ghasemi Y.
A summary of the article is shown below:
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 maximum nattokinase production and activity, the most enzyme activity reached to 883.371 U/ml when the media consisting of yeast extract (4.3804 g/L), tryptone (3.9899 g/L), K2HPO4 (1.6107 g/L) and CaCl2 (0.01 g/L). Entrapping the transformant cell in calcium alginate could gain more activity and lowering the media cost. The present study aim to produce the soluble nattokinase with high activity in the low costs of media that having high resistance against metal ions, detergents and organic solvents, which can easily use in medicines and detergents.
Check out the article’s website on Pubmed for more information:
This article is a good source of information and a good way to become familiar with topics such as:
Calcium alginate;E.coli BL21(DE3);Fibrinolytic;Industrial applications ;Nattokinase
Categories: Science News