Production of soluble human interleukin-6 in cytoplasm by fed-batch culture of recombinant E-coli

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The major objective of this study is to identify fed-batch culture conditions optimal for the production of human interleukin-6 (hIL-6) in a soluble form. Five different expression vectors were constructed for the expression of hIL-6 and hIL-6s fused with NusA, maltose binding protein (MBP), thioredoxin (Trx) or ubiquitin (Ubi). A series of flask cultures were conducted in LB medium at 37 degrees C. The intact hIL-6 was expressed mostly in the form of inclusion body. More than 95% of the hIL-6 fused with NusA (NusA/hIL-6) and about 90% of MBP/hIL-6 were expressed in a soluble form, whereas Trx/hIL-6 and Ubi/hIL-6 were expressed mostly in the form of inclusion body. Based on this result, NusA was selected as the fusion partner for the production of hIL-6 in the subsequent experiments. A series of pH-stat fed-batch cultures of an E. coli BL21(DE3) transformed with a NusA/hIL-6 expression vector were conducted in a bioreactor with a working volume of about 3 L. As the amount of nitrogen source was increased in the feeding medium, more soluble NusA/hIL-6 was produced, while the total amount was not significantly changed. Under the best conditions tested, about 90% of NusA/hIL-6 was produced in the soluble form. In this case, the concentration of soluble NusA/hIL-6 was 7.5 g/L with a volumetric productivity of 0.43 g/L-h.
Publisher
AMER CHEMICAL SOC
Issue Date
2005-03
Language
English
Article Type
Article
Keywords

HIGH-CELL-DENSITY; SECRETORY PRODUCTION; EXPRESSION; GROWTH; PURIFICATION; PROTEINS; FERMENTATION; CULTIVATION; STRAINS; CLONING

Citation

BIOTECHNOLOGY PROGRESS, v.21, no.2, pp.524 - 531

ISSN
8756-7938
DOI
10.1021/bp049645j
URI
http://hdl.handle.net/10203/88850
Appears in Collection
CBE-Journal Papers(저널논문)
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