---

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.