Welcome to International Network for Natural Sciences | INNSpub

Paper Details

Research Paper | July 1, 2015

VIEWS 1
| Download 4

Enhanced production of human epidermal growth factor (EGF) in Escherichia coli

Mohamad Mehdi Namvaran, Valiollah Babaeipour, Hosein Vahidi, Javad Ranjbari, Fatemeh Abarghooi Kahaki, Firouz Ebrahimi

Key Words:


Int. J. Biosci.7(1), 30-37, July 2015

DOI: http://dx.doi.org/10.12692/ijb/7.1.30-37

Certification:

IJB 2015 [Generate Certificate]

Abstract

Epidermal Growth Factor (EGF) is a monomer polypeptide consists of 53 amino acids with three intra-molecular disulfide bridges bonds. EGF is produced from different tissues in human and possesses valuable therapeutic effects. In the present study, we reported the high throughput expression of hEGF in Escherichia coli BL21 (DE3). For achieving this purpose, the designed vector was constructed based upon pET-28a (+) with T7 promoter. The synthetic hEGF gene was cloned in pET28a using NcoI and HindIІІ sites. Recombinant vector, pET28-hEGF, was transferred into E. coli BL21 (DE3) and induced for expression in a lab and bench scale. For this reason, at first effects of medium, temperature, and induction time in three-level were investigated on production enhancement of hEGF by using Taguchi experimental design in shake flask. Data processing by Qualitek-4 software was shown that maximum production acquires from TB medium at 28°C with IPTG concentration of 0.1mM. Under these conditions the final cell dry weight and the ultimate of hEGF concentration were 5.61g/l and 1.094 g/l respectively. Then, effects of induction time and glycerol concentration were examined at three levels in a bench bioreactor with full factorial approach. Under optimized conditions glycerol 15 g/l and induction time at OD600=5 with medium TB obtained final cell dry weight and the last of hEGF concentration were 10.58g/l and 2.28 g/l respectively. This amount of protein is one of the highest values which have been reported in non-continuous system.

VIEWS 1

Copyright © 2015
By Authors and International Network for
Natural Sciences (INNSPUB)
http://innspub.net
This article is published under the terms of the Creative
Commons Attribution Liscense 4.0

Enhanced production of human epidermal growth factor (EGF) in Escherichia coli

Babaeipour V, Shojaosadati SA, Maghsoudi N. 2013. Maximizing production of human interferon-γ in HCDC of recombinant E. coli. Iranian Journal of Pharmacutical Research 12, 563-572.

Baneyx F. 1999. Recombinant protein expression in Escherichia coli. Current Opinion in Biotechnology. 10, 411-421.

Barrow RE, Wang CZ, Evans MJ, Herndon DN. 1993. Growth factors accelerate epithelial repair in sheep trachea. Lung 171, 335–344. http://dx.doi.org/10.1007/BF00165699

Chem R. 2011. Bacterial expression systems for recombinant protein production: E. coil and beyond. Biotechnology Advances 30, 1102-1107. http://dx.doi.org/10.1016/j.biotechadv.2011.09.013.

Chia CM, Winston RM, Handyside AH. 1995. EGF, TGF-alpha and EGFR expression in human preimplantation embryos. Development 121, 299– 307.

Choi JH, Keum KC, Lee SY. 2006. Production of recombinant proteins by high cell density culture of Escherichia coli. Chemical Engineering Science 61, 876-885. http://dx.doi.org/10.1186/1475-2859-5-S1-P86

Cunningham F, Deber CM. 2007. Optimizing synthesis and expression of transmembrane peptides and proteins. Methods 41, 370–380. http://dx.doi.org/10.1016/j.ymeth.2006.07.003

Dreux AC, Lamb DJ, Modjtahedi H, Ferns GA. 2006. The epidermal growth factor receptors and their family of ligands: their putative role in atherogenesis. Atherosclerosis 186, 38–53. http://dx.doi.org/10.1016/j.clp.2004.03.015.

Groenen LC, Nice EC, Burgess AW. 1994. Structure-function  relationships  for  the  EGF/TGF-alpha family of mitogens. Growth factors 11, 235–257. http://dx.doi.org/10.3109/08977199409010997

Goldman B, Mach A, Wurzel J. 1996. Epidermal growth factor promotes a cardiomyoblastic phenotype in human fetal cardiac myocytes. Exp. Cell Research 228, 237–245. doi: http://dx.doi.org/10.1186/1475-2840-8-43

Gordon E, Horsefield R, Swarts HG, De Pont JJ, Neutze R, Snijder A. 2008. Effective high-throughput over production of membrane proteins in Escherichia coli. Protein Expression and Purification 62, 1 –8. http://dx.doi.org/10.1016/j.pep.2008.07.005.

Gupta C. 1996. The role of epidermal growth factor receptor (EGFR) in male reproductive tract differentiation: stimulation of EGFR expression and inhibition of Wolffian duct differentiation with anti-EGFR antibody. Endocrinology 137, 905–910. http://dx.doi.org/10.1210/en.

Harris RC, Chung E, Coffey RJ. 2003. EGF receptor ligands. Experimental Cell Research 284, 2– 13. http://dx.doi.org/10.1.1.457.4748&rep.

Lee SY. 1996. High cell density cultivation of Escherichia coli. Trends in Biotechnology 14, 98–105. http://dx.doi.org/10.1186/1475-2859-9-42.

Liu A, Davis RJ, Flores C, Menon M, Seethalakshmi L. 1992. Epidermal growth factor:receptor  binding  and  effects  on  the  sex  accessory organs  of  sexually  mature  male  mice.  Journal of Urology 148, 427–431. http://dx.doi.org/10.1093/humrep/des360

Lu HS, Chai JJ, Li M, Huang BR, He CH, Bi RC. 2001. Crystal structure of human epidermal growth factor and its dimerization. Journal of Biological Chemistry 276, 34913-34917. http://dx.doi.org/10.1016/S0092-8674(02)00963-7.

Kahaki AF, Babaeipour V, Rajabi MH, Mofid MR. 2014. High Overexpression and Purification of Optimized Bacterio-Opsin from Halobacterium Salinarum R1 in E. coli, Applied Biochemistry and Biotechnology 174, 1558-1571. http://dx.doi.org/10.1007/s12010-014-1137-2.

Kato M, Mizuguchi M, Takashima S. 1995. Developmental changes of epidermal growth factor-like immunoreactivity in the human fetal brain. Journal of Neurosciences Research 42, 486–492. http://dx.doi.org/10.1002/jnr.49042040

Klepsch MM, Persson JO, De Gier JL. 2011. Consequences of the over expression of a eukaryotic membrane protein, the human KDEL receptor, in Escherichia coli. Journal of Molecular Biology 407, 532 – 542. http://dx.doi.org/10.1016/j.jmb.

Ogiso H, Ishitani R, Nureki O. 2002. Crystal structure of the complex of human epidermal growth factor and receptor extracellular domains. Cell 110, 775–787.

Singh AB, Mukherjee K. 2013. Supplementation of substrate uptake gene enhances the expresson of rhIFN-β in high cell density fed-batch cultures of E. coli. Molecular Biotechnology 54, 692–702. http://dx.doi.org/10.1007/s12033-012-9611-y.

Stahlman MT, Orth DN, Gray ME. 1989. Immunocytochemical localization of epidermal growth factor in the developing human respiratory system and in acute and chronic lung disease in the neonate. Laboratory Investigation 60, 539–547. http://dx.doi.org/10.1203/00006450-199603000.

Thesleff I, Vaahtokari A, Partanen AM. 1995. Regulation of organogenesis. Common molecular mechanisms regulating the development of teeth and other organs. International Journal of Developmental Biology 39, 35–50. http://dx.doi.org/10.1007/s12011-014-0175-5

Thesleff I, Viinika L, Saxén L, Lehtonen E, Perheentupa J. 1988. The parotid gland is the main source of human salivary epidermal growth factor. Life Science, U. S. A 43, 13-18.

Van Zoelen EJ, Stortelers C, Lenferink AE,Van De Poll. 2000. The EGF domain: Requirements for binding to receptors of the ErbB family. Vitamins and Hormones 59, 99–131. http://dx.doi.org/10.1080/08977190902891010.

Venturi S, Venturi M. 2009. Iodine in evolution of salivary  glands  and  in  oral  health.  Nutrition  and Health 20, 119–134. http://dx.doi.org/10.1177/026010600902000204.

SUBMIT MANUSCRIPT

Style Switcher

Select Layout
Chose Color
Chose Pattren
Chose Background