METHOD FOR HIGH CONCENTRATION CULTIVATION OF RECOMBINANT ESCHERICHIA COLI CONTAINING GENE ENCODING T-20 PEPTIDE BY USING FED-BATCH CULTIVATION TECHNIQUE AND METHOD FOR SEPARATING AND PURIFYING THE T-20 PEPTIDE THEREFROM

摘要

PURPOSE: A method for high concentration cultivation of a recombinant Escherichia coli containing a gene encoding a T-20 peptide and a method for separating and purifying the T-20 peptide therefrom are provided, which high concentration cultivation of recombinant E. coli is accomplished by using a fed-batch cultivation technique, so that the T-20 peptide can be easily mass-produced. CONSTITUTION: The method for high concentration cultivation of a recombinant Escherichia coli containing a gene encoding a T-20 peptide comprises the steps of: (1) batch cultivating the recombinant E. coli containing the T-20 peptide gene in a cultivation medium of pH 6.8 to 7.2, added by a mineral solution; (2) repeatedly adding a nutrition medium into the cultivation medium without a change of pH in the cultivation medium during the cultivation process; and (3) adding lactose into the cultivation medium when the absorbance(OD) of the cultivation medium at 60nm is 30 to 50, and further cultivating the recombinant E. coli, wherein the recombinant E. coli is E. coli BmG3(KCCM 10506). The method for separating and purifying the T-20 peptide comprises destroying the cultured E. coli BmG3(KCCM 10506), and subjecting the destroyed biomass of E. coli BmG3(KCCM 10506) to C18 chromatography or Sepharose column using ketone solvent such as acetone.