1Department of pharmaceutical biotechnology, Faculty of pharmacy, Shahid Beheshti University of medical sciences, Tehran, Iran
2Department of Bioscience and Biotechnology, Malek Ashtar University of technology, Tehran, Iran
31-Department of pharmaceutical biotechnology, Faculty of pharmacy, Shahid Beheshti University of medical sciences, Tehran, Iran
4Department of pharmaceutics, Faculty of pharmacy, Shahid Beheshti University of medical sciences, Tehran, Iran
5Department of Biochemistry, Faculty of pharmacy, Isfahan University of medical sciences, Isfahan, Iran
6Department of Life Science Engineering, Faculty of New Science and Technologies, University of Tehran, Iran,
Abstract
Abstract Human insulin-like growth factor I (hIGF-I) is a kind of growth factor with clinical significance in medicine. The major objective of this study is over- production of recombinant human insulin-like growth factor I( rhIGF-I) through a developed process by recruiting effective factors in order to achieve the most recombinant protein. Up to now E. coli expression system has been widely used as a host to produce rhIGF-1 with high yields. Batch cultures as non-continuous fermentation were carried out to overproduce rhIGF-I in E. coli. The effects of culture medium type, induction temperature and amount of inducer on cell growth and IGF-I production were investigated in shaking flask. Taguchi design of experiments (DOE) method was used as the statistical method. Analysis of experimental data showed that maximum production of rhIGF-I was occurred in 32y culture medium at 28°C and 0.05 Mm IPTG. Under this condition, 0.7 g/L of rhIGF-I was produced as the inclusion bodies. Following optimization of these three factors, we have also optimized the amount of glucose and induction time in 5 liter top bench bioreactor. Full factorial design of experiment method was used for these two factors as the statistical method. 10 g/l and OD600=5 were selected as the optimum point of Glucose amount and induction time, respectively. Finally we have had 1.26 g/l rhIGF-1 production as the final product that is one of the best reported amounts.
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