Production characteristics of interferon-alpha using an L-arabinose promoter system in a high-cell-density culture.
Using high-cell-density culture of Escherichia coli under the control of an L-arabinose promoter (ParaB), several factors affecting the production of recombinant protein and the formation of inclusion bodies were studied. The inducer, L-arabinose, showed a maximal induction level above 10.7 mM in the final concentration. The concentration of inducer also affected the partition of interferon-alpha (IFN-alpha) into the soluble form and inclusion bodies. Induction kinetics of the rate of accumulation of IFN-alpha on the ParaB promoter showed a slower rate than those of other promoter systems, for example T7, lac or tac. These innate characteristics of ParaB enabled cells to grow continuously in spite of the metabolic burden induced by the expression of foreign protein. The duration time of induction could control the expression of both soluble and insoluble protein. The ratio of yeast extract to glycerol (N/C ratio) in feeding media significantly affected both the production level of recombinant protein and inclusion body formation. The reason for decreasing specific bioactivity during induction can be explained by the increased proportion of inclusion bodies in the total expressed IFN-alpha.
Lim
H K
HK
Mogam Biotechnology Research Institute, Yongin-City, Kyounggi-Do, Korea.
Jung
K H
KH
Park
D H
DH
Chung
S I
SI
D000818:Animals; D001089:Arabinose / Q000235:genetics* / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D003470:Culture Media / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D005990:Glycerol; D002479:Inclusion Bodies / Q000378:metabolism; D016898:Interferon-alpha / Q000096:biosynthesis* / Q000235:genetics; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis; D013997:Time Factors
D003470:Culture Media; D016898:Interferon-alpha; D011994:Recombinant Proteins; D001089:Arabinose; D005990:Glycerol