Functional Expression of FIP-<i>gts</i>, a Fungal Immunomodulatory Protein from<i>Ganoderma Tsugae</i>in Sf21 Insect Cells

  • JINN Tzyy-Rong
    Graduate Institute of Biotechnology, National Chung Hsing University Biopesticide Department, Taiwan Agricultural Chemicals and Toxic Substances Research Institute
  • WU Chi-Ming
    Biopesticide Department, Taiwan Agricultural Chemicals and Toxic Substances Research Institute
  • TU Wu-Chun
    Department of Entomology, National Chung Hsing University
  • KO Jiunn-Liang
    Institute of Medical and Molecular Toxicology, Chung Shan Medical University
  • TZEN Jason T. C.
    Graduate Institute of Biotechnology, National Chung Hsing University

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  • Functional Expression of FIP-gts, a Fungal Immunomodulatory Protein from Ganoderma Tsugae in Sf21 Insect Cells

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The mushrooms of diverse Lingzhi species have been traditionally consumed as luxurious functional food supplements in Chinese society. FIP-gts, a fungal immunomodulatory protein found in Song-Shan Lingzhi (Ganodera tsugae) has been proposed to possess therapeutic effects on cancer and autoimmune diseases. To produce active FIP-gts for evaluation of oral administration, a recombinant FIP-gts (rFIP-gts) fused with a 6His-tag at its C-terminus was expressed in Sf21 insect cells by the baculovirus expression system. High yield (about 70%) and purity (about 90%) of rFIP-gts was obtained by one-step nickel-affinity chromatography. The correctness of the harvested rFIP-gts was verified by Western blot and MALDI-MS analyses. Optimal expression of rFIP-gts was observed when the Sf21 cells were infected with multiplicity of infection of 10 for 72 h, and the yield was up to 47.2 μg/3×106 infected cells. The immunomodulatory activity of the purified rFIP-gts was detected as the induction of interleukin 2 released from murine splenocytes. Compared with the rFIP-gts produced in Escherichia coli cells, the rFIP-gts produced in Sf21 cells possessed evidently higher specific immunomodulatory activity.

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