en 23407680 Background: Bone Morphogenetic Proteins (BMPs) belong to the transforming growth factor-β (TGF-β) superfamily, and play an important role in bone metabolism. Recombinant forms of BMP-2 and BMP-7 are the only BMPs used clinically. In this study the mature part of human bone morphogenetic protein-7 (BMP-7) was engineered through substitution of the BMP-7 N-terminal sequence by heparin-binding site of BMP-2. This targeted substitution was made to enhance the binding affinity of the novel protein to the extracellular matrix components such as heparin and heparan sulfate proteoglycans (HSPGs). Methods: The engineered protein was expressed in Escherichia coli (E.coli). The PelB signal sequence was used to translocate soluble pro¬teins into the periplasmic space of E.coli. The protein was purified from periplasmic extract using Ni-NTA chromatography and the SDS-PAGE and western blot analysis confirmed the successful expression of the novel protein. Results: The novel hBMP-7 mutant was produced as approximately 16 kDa monomer. It was found that the heparin binding of this protein was approximately 50% more than that of the wild-type at a protein concentration of 500 ng/ml. Conclusion: The findings showed that the periplasmic expression may be suitable to produce complex proteins like BMPs. Bone morphogenetic protein-7 (BMP-7), Escherichia coli, Periplasmic expression 178 185 https://www.ajmb.org/En/Article.aspx?id=97 https://www.ajmb.org/PDF/En/FullText/97.pdf LeilaNematollahiMedical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran379VahidKhalajMedical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran135Seyedeh MalihehBabazadehDepartment of Microbiology, Islamic Azad University of Pharmaceutical Sciences, Tehran, Iran381AzamRahimpourMedical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran382HodaJahandarMedical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran383FatemehDavamiMedical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran130FereidounMahboudiMedical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran128