In silico analysis of different signal peptides for the secretory production of recombinant human keratinocyte growth factor in Escherichia coli |
| |
Affiliation: | 1. Department of Molecular Medicine, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran;2. Molecular Parasitology Laboratory, Pasteur Institute of Iran, Tehran, Iran;3. National Cell Bank, Pasteur Institute of Iran, Tehran, 1316943551 Iran;1. School of Biotechnology and Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China;2. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China;3. The 2011 Synergetic Innovation Center of Food Safety and Nutrition, Jiangnan University, Wuxi, 214122, China;4. Suqian Industrial Technology Research Institute of Jiangnan University, Suqian, 223814, China;1. Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran;2. Biochemistry Research Laboratory, Department of Chemistry, Sharif University of Technology, Tehran, Iran |
| |
Abstract: | BackgroundThe recombinant human truncated Keratinocyte growth factor (Palifermin) is the only FDA approved medicine for the treatment of oral mucositis. The Keratinocyte growth factor is a fairly unstable protein due to its high aggregation propensity and therefore its expression as a secretory protein may results in the production of a protein with more stability, higher solubility, better folding, enhanced biological activity, N-terminal authenticity and simplified downstream processing.ObjectiveThe aim of this study was in silico evaluation of 31 different secretory signal peptides to determine the best theoretical candidates for the secretory production of recombinant truncated human KGF in E. coli.MethodsThirty different prokaryotic signal peptides experimentally shown to be capable of recombinant protein secretion in E.coli, along with the native KGF signal peptide were selected for further investigations. The signal peptide sequences were retrieved from the UniProt database. The ability of SPs to act as a secretory leader peptide for rhKGF and the location of cleavage sites were predicted by SignalP 4.1. Physicochemical properties of the signal peptides, which may influence protein secretion, were analyzed by ProtParam and PROSOII. Furthermore, the mRNA secondary structure and Gibbs free energy profile of the selected SPs were analyzed in the fusion state with the rhKGF using Visual Gene Developer package.Results and ConclusionComputational analysis of the physicochemical properties affecting protein secretion identified Sec-B dependent OmpC, Bla, and StaI and SRP dependent TolB signal peptides as the best theoretical candidates for the secretory production of recombinant truncated human KGF in E.coli. |
| |
Keywords: | Signal peptides Secretory production Human keratinocyte growth factor |
本文献已被 ScienceDirect 等数据库收录! |
|