KMID : 0380020060210030204
|
|
Korean Journal of Biotechnology and Bioengineering 2006 Volume.21 No. 3 p.204 ~ p.211
|
|
Expression and Purification of Recombinant Human Interferon-gamma Produced by Escherichia coli
|
|
Park Jung-Ryeol
Kim Sung-Woo Kim Jae-Bum Jung Woo-Hyuk Han Myung-Wan Jo Young-Bae Jung Joon-Ki
|
|
Abstract
|
|
|
For the production of the recombinant human interferon-gamma (rhIFN-¥ã) in Escherichia coli, human glucagon and ferritin heavy chain were used as fusion partners. Even though rhIFN-¥ã is expressed as an inclusion body form in E. coli because of strong hydrophobicity of itself, over 50% of fused rhIFN-¥ã was expressed as soluble form in E. coli Origami(TM) (DE3) harboring pT7FH(HE)-IFN-¥ã which encodes ferritin heavy chain-fused rhIFN-¥ã. In the case of using glucagon-ferritin heavy chain hybrid mutant as a fusion partner, 6X His-tag was additionally introduced to N-terminus of GFHM(HE)-IFN-¥ã for enhancing purification yields of rhIFN-¥ã. Fusion protein HGFHM(HE)-IFN-¥ã with two 6X His-tag was more effectively bound to Ni-NTA agarose bead than GFHM(HE)-IFN-¥ã with a 6X His-tag. rhIFN-¥ã was completely purified from enterokinase-treated HGFHM(HE)-IFN-¥ã by Ni-NTA affinity column. For high-level production of rhIFN-¥ã, glucose was used as the sole carbon source with simple exponential feeding rate (2.4~7.2 g/h) in fed-batch process. The effective lactose concentration for the expression of the rhIFN-¥ã was 10~20 mM. Under the fed-batch culture conditions, rhIFN-¥ã production yield reached 11 g DCW/L for 6 hours after lactose induction.
|
|
KEYWORD
|
|
Recombinant human interferon-gamma, expression, purification, glucagon, ferritin heavy chain
|
|
FullTexts / Linksout information
|
|
|
|
Listed journal information
|
|
|