Volume 8, Issue 2 (2017)                   JMBS 2017, 8(2): 53-63 | Back to browse issues page

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Gholami tilko P, Hajihassan Z, Nazari N, Moghimi H. Optimization of the effective factors in E.coli growth producing recombinant β-NGF using response surface methodology. JMBS 2017; 8 (2) :53-63
URL: http://biot.modares.ac.ir/article-22-3829-en.html
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Abstract:   (11112 Views)
Production of recombinant proteins in Escherichia coli has been very common in recent decades. Many studies and experiments have been done in order to optimize the production and expression of recombinant proteins in E.coli. One strategy is using high cell density to increase recombinant protein production such as β-NGF in the cell. Therefore, in this study for the first time bacterial cell culture in high cell density was done using glycerol and yeast extract as carbon and nitrogen sources and MgCl2 as a growth effective factor. Also the effects of overnight culture conditions on bacterial growth were evaluated. Meanwhile culture conditions were optimized using response surface methodology (RSM) and the optimum conditions were as follows: 18/23 g/lit glycerol, 14.44 g/lit yeast extract and 10mM MgCl2. Also the obtained results indicated that the 14 hours incubation at 37 °C and 180 rpm were optimum conditions for the overnight culture. Our results showed that the rate of cell growth and recombinant β-NGF production in optimized condition is significantly higher than in basic medium.
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Article Type: _ | Subject: Agricultural Biotechnology
Received: 2015/12/28 | Accepted: 2017/09/23 | Published: 2018/01/27

1. Levi‐Montalcini, R. (1987) the nerve growth factor: 35 years later (Nobel lecture), Angewandte Chemie International Edition in English 26, 707-716.
2. Bax, B., Ferguson, G., Blaber, M., Sternberg, M. J., and Walls, P. H. (1993) Prediction of the three‐dimensional structures of the nerve growth factor and epidermal growth factor binding proteins (kallikreins) and an hypothetical structure of the high molecular weight complex of epidermal growth factor with its binding protein, Protein Science 2, 1229-1241
3. Sahdev, S., Khattar, S. K., and Saini, K. S. (2008) Production of active eukaryotic proteins through bacterial expression systems: a review of the existing biotechnology strategies, Molecular and cellular biochemistry 307, 249-264
4. Sun, W., Sun, C., Lin, H., Zhao, H., Wang, J., Ma, H., Chen, B., Xiao, Z., and Dai, J. (2009) The effect of collagen-binding NGF-β on the promotion of sciatic nerve regeneration in a rat sciatic nerve crush injury model, Biomaterials30, 4649-4656.
5. Fischer, W., Sirevaag, A., Wiegand, S. J., Lindsay, R. M., and Björklund, A. (1994) Reversal of spatial memory impairments in aged rats by nerve growth factor and neurotrophins 3 and 4/5 but not by brain-derived neurotrophic factor, Proceedings of the National Academy of Sciences 91, 8607-8611.
6. Jakubowska-Doğru, E., and Gümüşbaş, U. (2005) chronic intracerebroventricular NGF administration improves working memory in young adult memory deficient rats, Neuroscience letters 382, 45-50
7. Althaus, H. H. (2004) Remyelination in multiple sclerosis: a new role for neurotrophins?, Progress in brain research 146, 415-432
8. Heese, K., Low, J. W., and Inoue, N. (2006) Nerve growth factor, neural stem cells and Alzheimer’s disease, Neurosignals 15, 1-12
9. Kawamoto, K., and Matsuda, H. (2004) Nerve growth factor and wound healing, Progress in brain research 146, 369-384
10. Lambiase, A., Sacchetti, M., and Bonini, S. (2012) Nerve growth factor therapy for corneal disease, Current opinion in ophthalmology 23, 296-302
11. Bruce, G., and Heinrich, G. (1989) Production and characterization of biologically active recombinant human nerve growth factor, Neurobiology of aging 10, 89-94
12. Sahdev, S., Khattar, S. K., and Saini, K. S. (2008) Production of active eukaryotic proteins through bacterial expression systems: a review of the existing biotechnology strategies,Molecular and cellular biochemistry 307, 249-264
13. Wang, H., Xiao, Y., Fu, L., Zhao, H., Zhang, Y., Wan, X., Qin, Y., Huang, Y., Gao, H., and Li, X. (2010) High-level expression and purification of soluble recombinant FGF21 protein by SUMO fusion in Escherichia coli, BMC biotechnology 10, 14
14. Sarduy, E. S., Muñoz, A. C., Trejo, S. A., and Planes, M. d. l. A. C. (2012) High-level expression of Falcipain-2 in Escherichia coliby codonoptimization and auto-induction, Protein expression and purification 83, 59-69.
15. Williams, D. C., Van Frank, R. M., Muth, W. L., and Burnett, J. P. (1982) Cytoplasmic inclusion bodies in Escherichia coliproducing biosynthetic human insulin proteins, Science 215, 687-689
16. Lee, S. Y. (1996) Plastic bacteria? Progress and prospects for polyhydroxyalkanoate production in bacteria, Trends in Biotechnology 14, 431-438
17. Yee, L., and Blanch, H. (1992) Recombinant protein expression in high cell density fed-batch cultures of Escherichia coli, Nature Biotechnology 10, 1550-1556
18. Kramer, W., Elmecker, G., Weik, R., Mattanovich, D., and Bayer, K. (1996) Kinetic Studies for the Optimization of Recombinant Protein Formationa, Annals of the New York Academy of Sciences 782, 323-333
19. Alibolandi, M., and Mirzahoseini, H. (2011) Chemical assistance in refolding of bacterial inclusion bodies, Biochemistry research international 2011
20. Leandro, P., Lechner, M. C., de Almeida, I. T., and Konecki, D. (2001) Glycerol increases the yield and activity of human phenylalanine hydroxylase mutant enzymes produced in a prokaryotic expression system, Molecular genetics and metabolism 73, 173-178.
21. Savari, M., Esfahani, S. H. Z., Edalati, M., and Biria, D. (2015) Optimizing conditions for production of high levels of soluble recombinant human growth hormone using Taguchi method, Protein expression and purification 114, 128-135.
22. Sato, S., Ward, C. L., Krouse, M. E., Wine, J. J., and Kopito, R. R. (1996) Glycerol reverses the misfolding phenotype of the most common cystic fibrosis mutation, Journal of Biological Chemistry 271, 635-638.
23. Lin, E. (1976) Glycerol dissimilation and its regulation in bacteria, Annual Reviews in Microbiology 30, 535-578
24. Cao, W., Li, H., Zhang, J., Li, D., Acheampong, D. O., Chen, Z., and Wang, M. (2013) Periplasmic expression optimization of VEGFR2 D3 adopting response surface methodology: Antiangiogenic activity study, Protein expression and purification 90, 55-66
25. Shin, C. S., Hong, M. S., Bae, C. S., and Lee, J. (1997) Enhanced production of human mini‐proinsulin in fed‐batch cultures at high cell density of Escherichia coliBL21 (DE3)[pET‐3aT2M2], Biotechnology progress 13, 249-257
26. Lee, C., Sun, W., Burgess, B., Junker, B., Reddy, J., Buckland, B., and Greasham, R. (1997) Process optimization for large-scale production of TGF-α-PE40 in recombinant Escherichia coli: effect of medium composition and induction timing on protein expression, Journal of Industrial Microbiology and Biotechnology 18, 260-266
27. Nierhaus, KH. (2014) Mg2+, K+, and the Ribosome, Journal of Bacteriology 196, 3817–3819.
28. Vuillemin, M., Malbert, Y., Laguerre, S., Remaud-Siméon, M., and Moulis, C. (2014) Optimizing the production of an α-(1→2) branching sucrase in Escherichia coliusing statistical design, Applied microbiology and biotechnology 98, 5173-5184
29. Yang, Y., Zhang, D., Liu, S., Jia, D., Du, G., and Chen, J. (2012) Expression and fermentation optimization of oxidized polyvinyl alcohol hydrolase in E. coli, Journal of industrial microbiology & biotechnology 39, 99-104.
30. Yang, Y., Zhang, D., Liu, S., Jia, D., Du, G., and Chen, J. (2012) Expression and fermentation optimization of oxidized polyvinyl alcohol hydrolase in E. coli, Journal of industrial microbiology & biotechnology 39, 99-104.

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