Optimized Expression and Activity Assessment of Bacterial Staphylokinase in E. coli BL21(DE3) | ||
| Egyptian Journal of Medical Microbiology | ||
| Volume 34, Issue 3, July 2025 | ||
| Document Type: New and original researches in the field of Microbiology. | ||
| DOI: 10.21608/ejmm.2025.368878.1524 | ||
| Authors | ||
| Harith K. Buniya* 1; Omer D. Salahdin1; Abdulsalam N. Mohammed2 | ||
| 1Department of Biology, College of Education for Pure Science, University Of Anbar, Ramadi 31001, Al-Anbar, Iraq | ||
| 2Department of Medical Laboratory Techniques, College of Health and Technology, University of Al-Maarif, Ramadi 31001, Al-Anbar, Iraq | ||
| Abstract | ||
| Background: The application of recombinant proteins is rare following the high production costs of expressing proteins with expensive inducers, such as isopropyl-β-D-1-thiogalactopyranoside (IPTG). Staphylokinase (SAK), a fibrinolytic enzyme, is a small bacterial thrombolytic agent that specifically clots and converts plasminogens to plasmins and lysis fibrin clots. Objective: The primary aim of the present investigation is increasing the yield and lower the cost of staphylokinase production using Escherichia coli BL21 (DE3). Methodology: The influence of target protein in expression host by different culture medium, culture density, and IPTG concentration on the expression of SAK protein was explored. Results: The results indicated that only the culture density and concentration of IPTG were significant. This study achieved cost reduction by decreasing the IPTG inducer concentration (1.0 and 0.5mM), which acted as the inducer. The production rate was also maintained or increased in low culture density. Conclusion: Suitable production conditions, particularly diminished inducer concentration, effectively reduced upstream production costs and yielded high sak gene expression protein as an active form. | ||
| Keywords | ||
| Staphylokinase; E. coli; Protein expression; Culture density; IPTG | ||
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