METHOD OF FED-BATCH CULTIVATION AND RECOMBINANT PROTEIN EXPRESSION IN ESCHERICHIA COLI BL21(DE3) IN DEUTERATED MEDIUM

摘要

A method for cultivation of E.coli BL21 and its derivatives in deuterated media in fed-batch culture and expression of recombinant DNA products is described. The method is based on the usage of inexpensive deuterated carbon sources succinate and acetate together. The simultaneous supply of these substrates at molar ratio of 4-8 (mole/mole) acetate/succinate to the culture enables good growth to high cell densities and expression in fed-batch culture. A specific fed-batch control algorithm is applied to automatically adjust the feeding rate to the rate of the utilization of carbon sources based on dissolved oxygen control.