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.
Int J Mol Sci. 2013 Jan 24;14(2):2230-41. doi: 10.3390/ijms14022230.
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.
从金针菇（Flammulina velutipes）中分离得到的一种真菌免疫调节蛋白（FIP-fve）与免疫球蛋白重链的可变区具有结构相似性。在本研究中，在最佳培养条件（0.2 mM IPTG 和 28°C）下，在 transetta（DE3）中高表达了带有 N 端 His 标签的重组生物活性 FIP-fve 蛋白。通过在溶菌酶裂解过程中增加超声处理，提高了蛋白的纯化效率。生物活性 FIP-fve 蛋白的产量达到 29.1mg/L，纯度为 97.1%，具有大量的工业应用潜力。酶联免疫吸附试验显示，5mg/kg 体重的 His-FIP-fve 组小鼠血清中白细胞介素-2（IL-2）和γ干扰素（IFN-γ）的含量最大增加（p<0.01），与对照组相比，IL-4 的产生没有明显差异。
