Full factorial design experiments for investigating the effect of calculated amounts of amino acids on cell density and production of human interleukin-2 in recombinant E.coli BL21.

The expression of recombinant proteins can lead to the depletion of amino acid pools of the host. This phenomenon can have adverse effects on growth and production of the cell. The enrichment of culture medium with certain amounts of amino acids can be considered as an effective way to overcome this problem. In this study a stoichiometric model was used to estimate the necessary amounts of amino acids needed to enhance the production of human interleukin-2 by recombinant Escherichia coli BL21 [pET21a-rhil2]. The model avoids dealing with all the metabolic reactions by simply lumping them into a single reaction. The most important amino acids namely Leucine, Glutamine, Aspartic acid and Glycine were chosen for further investigations. Full factorial design (Yates table) was used in order to investigate the effect of the addition of predetermined amounts of amino acids on cell growth and production of human interleukin-2. Glutamine had the most promising effect on growth (OD600=3.76) which showed 50 percent increase comparing to the standard medium.

کلیدواژه ها:

Recombinant E.coli ، amino acid ، stoichiometric model ، full factorial design