title: Expression and Secretion of Endostar Protein by Escherichia Coli: Optimization of Culture Conditions Using the Response Surface Methodology.
abstract: Endostar as a specific drug in treatment of the nonsmall cell lung cancer is produced using Escherichia coli expression system. Plackett-Burman design (PBD) and response surface methodology (RSM) are statistical tools for experimental design and optimization of biotechnological processes. This investigation aimed to predict and develop the optimal culture condition and its components for expression and secretion of endostar into the culture medium of E. coli. The synthetic endostar coding sequence was fused with PhoA signal peptide. The nine factors involved in the production of recombinant protein-postinduction temperature, cell density, rotation speed, postinduction time, concentration of glycerol, IPTG, peptone, glycine, and triton X-100-were evaluated using PBD. Four significant factors were selected based on PBD results for optimizing culture condition using RSM. Endostar was purified using cation exchange chromatography and size exclusion chromatography. The maximum level of endostar was obtained under the following condition: 13.57-h postinduction time, 0.76 % glycine, 0.7 % triton X-100, and 4.87 % glycerol. The predicted levels of endostar was significantly correlated with experimental levels (R 2 = 0.982, P = 0.00). The obtained results indicated that PBD and RSM are effective tools for optimization of culture condition and its components for endostar production in E. coli. The most important factors in the enhancement of the protein production are glycerol, glycine, and postinduction time.
authors.affiliation: Tuberculosis and Lung Disease Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.|Department of Biotechnology, Zahravi pharmaceutical company, Tabriz, Iran.
authors.forename: Abbas
authors.initials: A
authors.lastname: Mohajeri
authors.affiliation: Drug Applied Research Center, Immunology Lab, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Jalal
authors.initials: J
authors.lastname: Abdolalizadeh
authors.affiliation: Tuberculosis and Lung Disease Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.|Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Younes
authors.initials: Y
authors.lastname: Pilehvar-Soltanahmadi
authors.affiliation: Department of Biotechnology, Zahravi pharmaceutical company, Tabriz, Iran.
authors.forename: Farhad
authors.initials: F
authors.lastname: Kiafar
authors.affiliation: Tuberculosis and Lung Disease Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. zarghami@tbzmed.ac.ir.|Drug Applied Research Center, Immunology Lab, Tabriz University of Medical Sciences, Tabriz, Iran. zarghami@tbzmed.ac.ir.|Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran. zarghami@tbzmed.ac.ir.
authors.forename: Nosratollah
authors.initials: N
authors.lastname: Zarghami
mesh_terms: D001431:Bacteriological Techniques; D002852:Chromatography, Ion Exchange; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D043169:Endostatins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005990:Glycerol / Q000494:pharmacology; D005998:Glycine / Q000494:pharmacology; D021382:Protein Sorting Signals / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D011994:Recombinant Proteins
chemical_list: D003470:Culture Media; D043169:Endostatins; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C522911:endostar protein; D005990:Glycerol; D005998:Glycine
keywords: Angiogenesis inhibitors; Endostar; Escherichia coli; Lung cancer