Optimization of soluble human interferon-γ production in Escherichia coli using SUMO fusion partner.
Interferon-γ (IFN-γ) is a broad-spectrum antiviral glycoprotein that produced by lymphatic T cells and natural killer cells those who had stimulated by antigen. Human IFN-γ (hIFN-γ) often used in clinical research and practice because of its bioactivity, for example, antivirus, antitumor, controlling cell apoptosis, and the strict selectivity. However, due to the difficulties of Escherichia coli expression system meet in protein folding, the hIFN-γ often existed as inclusion body. The production of soluble hIFN-γ can be developed to shorten the production cycle and decrease the cost. In this study, small ubiquitin-related modifier fusion technology was used to express and purify recombinant hIFN-γ. Expression induced by adding 50 mM arginine and 1 % (w/v) glycerol into the culture at 24 °C existed as a soluble form of 70 % in total protein. Finally, about 62 mg recombinant hIFN-γ was obtained from 1 L fermentation culture with no less than 96 % purity. Determined by cytopathic effect inhibition assay, the specific activity of the recombinant hIFN-γ achieved at 7.78 × 10(5) IU/mL.
Jiangsu Province Key Laboratory for Molecular and Medicine Biotechnology, College of Life Science, Nanjing Normal University, No. 1 Wenyuan Road, Nanjing 210046, Jiangsu, People's Republic of China.
Fenfen
F
Zhu
Qi
Q
Wang
Hefang
H
Pu
Shasha
S
Gu
Lan
L
Luo
Zhimin
Z
Yin
D000998:Antiviral Agents / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D025841:Small Ubiquitin-Related Modifier Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D014721:Vesicular stomatitis Indiana virus / Q000187:drug effects
D000998:Antiviral Agents; D011993:Recombinant Fusion Proteins; D025841:Small Ubiquitin-Related Modifier Proteins; D007371:Interferon-gamma