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Showing 2 results for Gholami tilko
Pouria Gholami tilko, Zahra Hajihassan, Navid Nazari, Hamid Moghimi,
Volume 8, Issue 2 (10-2017)
Abstract
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.
Z. Hajihassan, S.m. Sadat, P. Gholami tilko ,
Volume 10, Issue 1 (Winter 2019)
Abstract
Aims: Nerve growth factor (β-NGF) is an important therapeutic agent for the treatment of neurodegenerative diseases such as Alzheimer’s disease; so, recombinant production of it in industrial scale is of high importance. The aim of this study is to optimize the effective factors in achieving the highest rate of β-NGF protein production in the bioreactor.
Materials & Methods: As E. coli is a suitable host for industrial production of recombinant proteins, E. coli DE3 strain was used for production of recombinant β-NGF. Also, fermentation was performed in a 5-L bioreactor and % dissolved oxygen (%DO) and post-induction temperature values were optimized by response surface methodology (RSM). At first, the effects of these two variables on the level of total protein were studied. So, in every experiment, bacterial proteins were isolated and total protein concentration was determined by Bradford assay.
Findings: The results indicated that %DO and post-induction temperature of 30% and 28.5ºC were the best values for increased production of total protein; in these circumstances, total protein concentration was 9.6±0.61 mg/ml. Finally, the effects of these variables on recombinant β-NGF production were surveyed by dot blot analysis, indicating the maximum β-NGF expression level on the optimized condition.
Conclusion: In conclusion, %DO and post-induction temperature not only affect cell growth of recombinant E. coli, but also have a direct impact on recombinant protein expression and production, such as β-NGF.
