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Int. J. Mol. Sci. 2013, 14(2), 2230-2241; doi:10.3390/ijms14022230

High-Yield Production in Escherichia coli of Fungal Immunomodulatory Protein Isolated from Flammulina velutipes and Its Bioactivity Assay in Vivo

1 Key Laboratory of Saline-alkali Vegetation Ecology Restoration in Oil Field (SAVER), Ministry of Education, Alkali Soil Natural Environmental Science Center (ASNESC), Northeast Forestry University, Harbin Hexing Road, Harbin 150040, China 2 Institute of Microbiology, Heilongjiang Academy of Sciences, Harbin 150010, China 3 School of Physics, Nankai University, Tianjin 130071, China
* Author to whom correspondence should be addressed.
Received: 11 December 2012 / Revised: 8 January 2013 / Accepted: 18 January 2013 / Published: 24 January 2013
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A fungal immunomodulatory protein isolated from Flammulina velutipes (FIP-fve) has structural similarity to the variable region of the immunoglobulin heavy chain. In the present study, the recombinant bioactive FIP-fve protein with a His-tag in N-terminal of recombinant protein was expressed in transetta (DE3) at a high level under the optimized culturing conditions of 0.2 mM IPTG and 28 °C. The efficiency of the purification was improved with additional ultrasonication to the process of lysozyme lysis. The yield of the bioactive FIP-fve protein with 97.1% purity reached 29.1 mg/L with a large quantity for industrial applications. Enzyme-linked immunosorbent assay showed a maximum increase in interleukin-2 (IL-2) and gamma interferon (IFN-γ) for the mice serum group of 5 mg/kg body mass (p < 0.01) with three doses of His-FIP-fve. However, the production of IL-4 had no apparent difference compared to the control.
Keywords: protein expression; large-scale production; ELISA protein expression; large-scale production; ELISA
This is an open access article distributed under the Creative Commons Attribution License (CC BY) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Kong, X.; Zhang, J.; Han, X.; Zhang, P.; Dai, X.; Liu, J.; Zhang, X.; Lee, I.; Liu, S. High-Yield Production in Escherichia coli of Fungal Immunomodulatory Protein Isolated from Flammulina velutipes and Its Bioactivity Assay in Vivo. Int. J. Mol. Sci. 2013, 14, 2230-2241.

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