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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

Xianghui Kong ^1,2 , Jiechi Zhang ² , Xue Han ¹ , Piqi Zhang ² , Xiaodong Dai ² , Jianing Liu ² , Xinxin Zhang ¹ , Imshik Lee ^1,3  and Shenkui Liu ^1,*

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; in revised form: 8 January 2013 / Accepted: 18 January 2013 / Published: 24 January 2013

Download PDF Full-Text [665 KB, uploaded 24 January 2013 11:31 CET]

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

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