Extracellular expression and efficient purification of a functional recombinant Volvariella volvacea immunomodulatory protein (FIP-vvo) using Pichia pastoris system

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• 作者：Xilin Sun ; Wei Huang ; Sijia Xiao ; Chongyang Liang ; Shuqin Zhang ; Zhiyi Liu ; Fei Sun
• 关键词：reFIP-vvo ; Volvariella volvacea ; Pichia pastoris ; Immunomodulatory
• 刊名：Protein Expression and Purification
• 出版年：February, 2014
• 年：2014
• 卷：94
• 期：Complete
• 页码：95-100
• 全文大小：962 K

文摘

The fungal immunomodulatory proteins (FIPs) are a new protein family identified from several edible and medical mushrooms and play an important role in antitumor, anti-allergy and immunomodulating activities. A gene encoding the FIP-vvo was cloned from the mycelia of Volvariella volvacea and recombinant expressed in the Pichia pastoris expression system. SDS-PAGE, amino acid composition and circular dichroism analyses of the recombinant FIP-vvo (reFIP-vvo) indicated that the gene was correctly and successfully expressed. In vitro assays of biological activities revealed that the reFIP-vvo exhibited similar immunomodulating capacities as native form. The reFIP-vvo significantly stimulated the proliferation of mouse spleen lymphocytes and apparently enhanced the expression level of IFN-纬 released from the mouse splenocytes. Taken together, the FIP-vvo gene from V. volvacea has been integrated into the yeast genome and expressed effectively at a high level (about 410 mg/L), it was capable of agglutinating sheep and rat red blood cells. The reFIP-vvo possessed very similar biological activities to native FIPs, suggesting its potential application as a food supplement or immunomodulating agent in pharmaceuticals and even medical studies.