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Effect of chitosan on production of insulin-like growth factor i protein in Escherichia coli

Javad Ranjbari, HamidrezaMoghimi, Hossein Vahidi, Valiollah Babaeipour, Abbas Alibakhshi, Roghayeh Arezumand

Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, ShaheedBeheshti University of Medical Sciences, Tehran, Iran

Students Research Committee, Faculty of Pharmacy, Shaheed Beheshti University of Medical Sciences, Tehran, Iran

Department of Life Science Engineering, Faculty of New Science and Technologies, University of Tehran, Iran

Department of Pharmaceutics, Faculty of Pharmacy, Shaheed Beheshti University of medical Sciences, Tehran, Iran,

Department of Medical Biotechnology, Faculty of medicin, Shaheed Beheshti University of medical Sciences, Tehran, Iran

Department of Molecular Medicine, Pasteur Institute of Iran, Tehran, Iran

Key words: RhIGF-I, Chitosan, E. coli, non- continuous fermentation, MIC.

Abstract

ecoliHuman insulin-like growth factor 1 (hIGF-1) is a kind of growth factor with clinical significance in medicine. 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 fermentations were carried out to overproduce rhIGF-I in E. coli. Chitosan and its derivatives, which are known to possess multiple functional properties, have attracted considerable interest due to their biological activities and potential applications in the pharmaceutical, food, agricultural and environmental industries. Many researchers have focused on Chitosan as a potential source of bioactive material in the past few decades. This study focuses on the Chitosan activity on increasing the cell growth and rh-IGF-1concentration in batch culture by affecting on food uptake. The major objective of this study is over- production of recombinant human insulin-like growth factor 1(rhIGF-1) through a developed process by Chitosan as aneffective factor, in order to achieve a higher recombinant protein. According to our previous study analysis of experimental data showed that maximum production of rhIGF-I (1.26 g/l) was occurred in 32y culture medium at 32°C with 0.05 Mm IPTG as inducer and 10 g/l glucose concentration. Under this condition, we have also optimized the amount of Chitosan. Regarding to Minimum Inhibitory Concentration (MIC) of Chitosan (2 μg/ml) on E.coli (B/DE3), three concentrations of Chitosan (0.5, 1 and 1.5 μg/ml) were selected. Finally, 1.5μg/ml was selected as the optimum point of Chitosan amount and we reached to a concentration of 1.51 g/l rhIGF-1 at this point.

Source/In English: http://www.innspub.net/wp-content/uploads/2015/01/IJB-V6No2-p172-179.pdf

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