Jafari R, Mirshahi M,
Volume 65, Issue 1 (3-2008)
Abstract
Background: Streptokinase (SK) is an effective and specific thrombolytic treatment of
acute myocardial infarction. Despite its significant limitations, streptokinase remains the
drug of choice particularly in countries with poorer economies because of its relatively
low cost. In this study, the production and purification of streptokinase using a pMAL
expression vector were evaluated.
Methods: The pMAL vector, including the skc gene, obtained from Streptococcus
equisimilis H46A, was transformed into E. coli BL21 to produce the soluble active fusion
protein SK-MBP. The conditions of SK production were optimized by manipulating
temperature, induction time and IPTG concentrations. This protein was purified by
DEAE-sepharose column chromatography and the final purity was determined and
activity of purified SK-MBP was measured using a synthetic substrate (S2251).
Results: After optimizing the production conditions, SK-MBP was the major portion of
total protein. Purified SK-MBP formed a single band using SDS-PAGE and had high
biological activity.
Conclusion: In this study we used pMAL expression vector to produce SK-MBP in E.
coli BL21. Using this method we prevented the accumulation of inclusion bodies in spite
of the high level of production of SK-MBP. Choosing a suitable host organism for the
production of recombinant proteins is one of the most important factors that influence the
level of desired protein production. Further studies are recommended to test other host
organisms for this purpose.
