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標題: Functional expression of FIP-gts, a fungal immunomodulatory protein from Ganoderma tsugae in Sf21 insect cells
作者: Jinn, T.R.
Wu, C.M.
Tu, W.C.
Ko, J.L.
Tzen, J.T.C.
關鍵字: baculovirus expression system;fungal immunomodulatory protein;interleukin 2;lingzhi;nickel-affinity chromatography;ling zhi-8 lz-8;amino-acid-sequence;escherichia-coli;baculovirus;lymphocytes;mushroom;system;mice;proliferation;lucidium
Project: Bioscience Biotechnology and Biochemistry
期刊/報告no:: Bioscience Biotechnology and Biochemistry, Volume 70, Issue 11, Page(s) 2627-2634.
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 mu g/3 x 10(6) 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.
ISSN: 0916-8451
DOI: 10.1271/bbb.60232
Appears in Collections:生物科技學研究所

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