Introduction: Osteoporosis is a common disease in the world, which is prevalent especially in aging societies. Teriparatide is a synthetic peptide amino acids 1-34 of the N-terminal of human parathyroid hormone (PTH) and approved by FDA in 2002 for treatment of osteoporosis. Ssp DnaB mini-intein with 154 amino acids is a protein which has self-splicing property. It acts as a fusion tag/fusion partner for expression of peptides and proteins in biological tools. Bioseparation technique is mediated by self-splicing inteins has become an excellent tool for affinity tag-based protein purification techniques and an alternative to conventional splicing by site-specific endoproteases. In this study the effects of Ssp DnaB mini-intein on expression and solubility of Teriparatde peptide during expression in E. coli was evaluated.
Methods: The fusion of Ssp DnaB mini-intein and Teriparatide encoding sequences was achieved through the PCR technique. The fusion gene was then cloned in pET28a plasmid containing a N-terminal Histidin-tag. Subsequently the construct was introduced into E. coli BL21 strain through the process of transformation. The E. coli cells was then grown in Luria Broth (LB) medium supplemented with kanamycin antibiotic to produce the desired recombinant protein. Upon completing the solubilization process of the protein, the intein and peptide are separated under certain conditions (pH, temperature) and purified, resulting in the successful production of the desired peptide.
Results: The finding of this study revealed the potential of Ssp DnaB mini-intein in preparation of Teriparatide in E. coli and the expression and solubility of peptide be in acceptable level.
Conclusion: Using the inteins facilitate the purification of peptides and proteins without need to endoproteases therefore this fusion tag can be used for producing this peptide in industry scale in easier way.
Keywords: Teriparatide, Ssp DnaB mini-intein, Expression, E. coli