title: Designing novel construction for cell surface display of protein E on Escherichia coli using non-classical pathway based on Lpp-OmpA.
abstract: Today, transference of recombinant protein on the outer surface of bacteria is deemed as a valuable process for various applications in biotechnology including preparation of vaccines. In this study, Lpp-OmpA structure was used to present outer membrane protein E of Haemophilus influenzae on E. coli outer membrane. Also, a structure was designed according to Lpp-OmpA based on non-classical secretion pathway using bioinformatics software such as MEMSAT-SVM, ScrotumP and SignalP where it lacked any signal peptide at its N-terminal. Potential of this structure in the presentation of protein E on the surface of E. coli through non-classical pathway was indicated by western blotting, SDS page and fluorescent microscopy techniques, similarly its effectiveness was compared with Lpp-OmpA system. The results of the current study showed that the new structure had higher efficiency than Lpp-OmpA, and it could transport protein E on outer membrane well. This study is the first report in the presentation of H. influenzae PE onto the surface of E. coli by Lpp-OmpA, and the structure originated from Lpp-OmpA, according to the non-classical secretion pathway. Our results suggest that non-classical secretion pathway may be exploited as a new secretory pathway on the outer surface of the cell for recombinant proteins.
authors.affiliation: Department of Biology, School of Basic Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors.forename: Meisam
authors.initials: M
authors.lastname: Jeiranikhameneh
authors.affiliation: Molecular Parasitology Laboratory, Department of Parasitology, Pasteur Institute of Iran, 69 Pasteur Avenue, Tehran, 1316943551, Iran. mrrazavi@pasteur.ac.ir.
authors.forename: Mohamad Reza
authors.identifier: http://orcid.org/0000-0002-2204-6267
authors.initials: MR
authors.lastname: Razavi
authors.affiliation: Department of Biology, School of Basic Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors.forename: Shiva
authors.initials: S
authors.lastname: Irani
authors.affiliation: Department of Tuberculosis and Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Seyed Davar
authors.initials: SD
authors.lastname: Siadat
authors.affiliation: Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mana
authors.initials: M
authors.lastname: Oloomi
keywords: Haemophilus influenza; Lpp-OmpA; Non-classical secretion pathway; Protein E; Surface display