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Tytuł:
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In silico analysis of different signal peptides for the secretory production of recombinant human keratinocyte growth factor in Escherichia coli.
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Autorzy:
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Dastjerdeh MS; Department of Molecular Medicine, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
Marashiyan M; Department of Molecular Medicine, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
Boroujeni MB; Department of Molecular Medicine, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
Golkar M; Molecular Parasitology Laboratory, Pasteur Institute of Iran, Tehran, Iran.
Shokrgozar MA; National Cell Bank, Pasteur Institute of Iran, Tehran, 1316943551 Iran.
Rahimi H; Department of Molecular Medicine, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran. Electronic address: .
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Źródło:
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Computational biology and chemistry [Comput Biol Chem] 2019 Jun; Vol. 80, pp. 225-233. Date of Electronic Publication: 2019 Mar 29.
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Typ publikacji:
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Journal Article
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Język:
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English
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Imprint Name(s):
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Publication: Oxford : Elsevier
Original Publication: Oxford : Pergamon, c2003-
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MeSH Terms:
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Computer Simulation*
Protein Sorting Signals*
Fibroblast Growth Factor 7/*chemistry
Recombinant Proteins/*chemistry
Amino Acid Sequence ; Escherichia coli/chemistry ; Fibroblast Growth Factor 7/genetics ; Humans ; Hydrophobic and Hydrophilic Interactions ; RNA, Messenger/chemistry ; Recombinant Proteins/genetics ; Solubility ; Thermodynamics
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Contributed Indexing:
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Keywords: Escherichia coli; Human keratinocyte growth factor; In silico; Secretory production; Signal peptides
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Substance Nomenclature:
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0 (FGF7 protein, human)
0 (Protein Sorting Signals)
0 (RNA, Messenger)
0 (Recombinant Proteins)
126469-10-1 (Fibroblast Growth Factor 7)
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Entry Date(s):
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Date Created: 20190419 Date Completed: 20190805 Latest Revision: 20190805
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Update Code:
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20240104
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DOI:
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10.1016/j.compbiolchem.2019.03.003
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PMID:
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30999249
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Background: The 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.
Objective: The 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.
Methods: Thirty 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 Conclusion: Computational 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.
(Copyright © 2019 Elsevier Ltd. All rights reserved.)
