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en Production of Recombinant Streptavidin and Optimization of Refolding Conditions for Recovery of Biological Activity زیست شناسی ملکولی Molecular Biology مقالات اصلی Original Article Background: Streptavidin is a protein produced by Streptomyces avidinii with strong biotin-binding ability. The non-covalent, yet strong bond between these two molecules has made it a preferable option in biological detection systems. Due to its extensive use, considerable attention is focused on streptavidin production by recombinant methods. Methods: In this study, streptavidin was expressed in Escherichia coli (E. coli) BL21 (DE3) pLysS cells and purified by affinity chromatography. Various dialysis methods were employed to enable the protein to refold to its natural form and create a strong bond with biotin. Results: Streptavidin was efficiently expressed in E. coli. Streptavidin attained its natural form during the dialysis phase and the refolded protein bound biotin. The addition of proline or arginine to the dialysis buffer resulted in a refolded streptavidin with greater affinity for biotin than refolding in dialysis buffer with no added amino acids. Conclusions: Dialysis of recombinant streptavidin in the presence of arginine or proline resulted in proper refolding of the protein. The recombinant dialyzed streptavidin bound biotin with affinity as great as that of a commercial streptavidin. Biotin, Protein Refolding, Streptavidin, Streptomyces 178 185 http://rbmb.net/browse.php?a_code=A-10-62-1&slc_lang=en&sid=1 Raoufe Modanloo Jouybari raoufemodanloo@yahoo.com 10031947532846006299 10031947532846006299 No Department of Medical Biotechnology, Faculty of Medicine, Arak University of Medical Sciences, Arak, Iran. Abdorrahim Sadeghi abdorrahims@gmail.com 10031947532846006300 10031947532846006300 No Endocrinology and Metabolism Research Center, Department of Biochemistry, School of Medicine, Arak University of Medical Sciences, Arak, Iran. Behzad Khansarinejad b_khansarinejad@yahoo.com 10031947532846006301 10031947532846006301 No Department of Microbiology and Immunology, Faculty of Medicine, Arak University of Medical Sciences, Arak, Iran. Shabnam Sadoogh Abbasian shabmicro@yahoo.com 10031947532846006302 10031947532846006302 No Molecular and Medicine Research Center, Department of Microbiology, School of Medicine, Arak University of Medical Sciences, Arak, Iran. Hamid Abtahi abtahi@arakmu.ac.ir. 10031947532846006303 10031947532846006303 Yes Molecular and Medicine Research Center, Department of Microbiology, School of Medicine, Arak University of Medical Sciences, Arak, Iran.