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

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

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Received: 11 December 2012; in revised form: 8 January 2013 / Accepted: 18 January 2013 / Published: 24 January 2013
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Abstract: 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 which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

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.

AMA Style

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. International Journal of Molecular Sciences. 2013; 14(2):2230-2241.

Chicago/Turabian Style

Kong, Xianghui; Zhang, Jiechi; Han, Xue; Zhang, Piqi; Dai, Xiaodong; Liu, Jianing; Zhang, Xinxin; Lee, Imshik; Liu, Shenkui. 2013. "High-Yield Production in Escherichia coli of Fungal Immunomodulatory Protein Isolated from Flammulina velutipes and Its Bioactivity Assay in Vivo." Int. J. Mol. Sci. 14, no. 2: 2230-2241.


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