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Production of Soluble Human Interleukin‐6 in Cytoplasm by Fed‐Batch Culture of Recombinant E. coli
Author(s) -
Kim Tae Wan,
Chung Bong Hyun,
Chang Yong Keun
Publication year - 2008
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp049645j
Subject(s) - recombinant dna , cytoplasm , escherichia coli , biology , microbiology and biotechnology , chemistry , biochemistry , gene
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 °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.

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