Method to improve expression of granulocyte colony stimulating factor by oxygenated DO-stat fed-batch culture

摘要

The present invention relates to a method for enhancing the expression of granulocyte colony stimulating factor (G-CSF) by oxygenated DO-stat fed-batch culture. More specifically, the present invention relates to DO-stat fed-batch culture of recombinant Escherichia coli transformed with G-CSF expression vector using L-arabinose operon of Salmonella strains. A method of maximizing efficiency and further enhancing expression of recombinant G-CSF from recombinant E. coli by inducing expression of the protein with L-arabinose at the time of induction of optimal expression, that is, at the end of batch culture and at the beginning of fed-batch culture. It is about. According to the present invention, even in the addition of oxygen, DO-stat fed-value cultivation, which is an indicator of the amount of dissolved oxygen, was efficiently performed, and was excellent in terms of cell mass, expression level of G-CSF, and stability of plasma. In addition, glucose added as a carbon source during the fed-batch culture was maintained below the minimum concentration of expression inhibition, and expressed a similar level of G-CSF as when using glycerol as a carbon source.