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-<I>gts</I>, a fungal immunomodulatory protein found in Song-Shan Lingzhi (<I>Ganodera tsugae</I>) has been proposed to possess therapeutic effects on cancer and autoimmune diseases. To produce active FIP-<I>gts</I> for evaluation of oral administration, a recombinant FIP-<I>gts</I> (rFIP-<I>gts</I>) 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-<I>gts</I> was obtained by one-step nickel-affinity chromatography. The correctness of the harvested rFIP-<I>gts</I> was verified by Western blot and MALDI-MS analyses. Optimal expression of rFIP-<I>gts</I> 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/3×10<SUP>6</SUP> infected cells. The immunomodulatory activity of the purified rFIP-<I>gts</I> was detected as the induction of interleukin 2 released from murine splenocytes. Compared with the rFIP-<I>gts</I> produced in <I>Escherichia coli</I> cells, the rFIP-<I>gts</I> produced in Sf21 cells possessed evidently higher specific immunomodulatory activity.
- Agricultural and Biological Chemistry
Agricultural and Biological Chemistry 70(11), 2627-2634, 2006-11-23
Japan Society for Bioscience, Biotechnology, and Agrochemistry