Enhanced production of human epidermal growth factor (EGF) in Escherichia coli
Paper Details
Enhanced production of human epidermal growth factor (EGF) in Escherichia coli
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.
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Mohamad Mehdi Namvaran, Valiollah Babaeipour, Hosein Vahidi, Javad Ranjbari, Fatemeh Abarghooi Kahaki, Firouz Ebrahimi , 2015. Enhanced production of human epidermal growth factor (EGF) in Escherichia coli. Int. J. Biosci., 7(1), 30-37.
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