Growth-associated synthesis of recombinant human glucagon and human growth hormone in high-cell-density cultures of Escherichia coli.
Synthesis of two recombinant proteins (human glucagon and human growth hormone) was investigated in fed-batch cultures at high cell concentrations of recombinant Escherichia coli. The glucose-limited growth was achieved without accumulation of metabolic by-products and hence the cellular environment is presumed invariable during growth and recombinant protein synthesis. Via exponential feeding in the two-phase fed-batch operation, the specific cell growth rate was successfully controlled at the desired rates and the fed-batch mode employed is considered appropriate for examining the correlation between the specific growth rate and the efficiency of recombinant product formation in the recombinant E. coli strains. The two recombinant proteins were expressed as fusion proteins and the concentration in the culture broth was increased to 15 g fusion growth hormone 1(-1) and 7 g fusion glucagon 1(-1). The fusion growth hormone was initially expressed as soluble protein but seemed to be gradually aggregated into inclusion bodies as the expression level increased, whereas the synthesized fusion glucagon existed as a cytoplasmic soluble protein during the whole induction period. The stressful conditions of cultivation employed (i.e., high-cell-density cultivation at low growth rate) may induce the increased production of various host-derived chaperones and thereby enhance the folding efficiency of synthesized heterologous proteins. The synthesis of the recombinant fusion proteins was strongly growth-dependent and more efficient at a higher specific growth rate. The mechanism linking specific growth rate with recombinant protein productivity is likely to be related to the change in cellular ribosomal content.
Shin
C S
CS
Bioprocess Engineering Laboratory, Hanhyo Institutes of Technology, Taejon, South Korea.
Hong
M S
MS
Kim
D Y
DY
Shin
H C
HC
Lee
J
J
D015169:Colony Count, Microbial; D004926:Escherichia coli / Q000254:growth & development / Q000378:metabolism*; D005934:Glucagon / Q000096:biosynthesis*; D013006:Growth Hormone / Q000096:biosynthesis*; D006801:Humans; D008828:Microbiological Techniques; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*
D011993:Recombinant Fusion Proteins; D013006:Growth Hormone; D005934:Glucagon