Functional Expression of FIP-gts, a Fungal Immunomodulatory Protein from Ganoderma Tsugae 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

Bibliographic Information

Other Title

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

Abstract

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.

Journal

• Bioscience, Biotechnology, and Biochemistry

  Bioscience, Biotechnology, and Biochemistry 70 (11), 2627-2634, 2006

  Japan Society for Bioscience, Biotechnology, and Agrochemistry

Keywords

• baculovirus expression system
• fungal immunomodulatory protein
• interleukin 2
• lingzhi
• nickel-affinity chromatography

Details

• CRID: 1390282681454600832
• NII Article ID: 10018524613
• NII Book ID: AA10824164
• DOI: 10.1271/bbb.60232
• ISSN: 13476947
• ISSN: 09168451
• NDL BIB ID: 8560301
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I8560301
• Web Site: http://www.tandfonline.com/doi/pdf/10.1271/bbb.60232
• Web Site: https://search.jamas.or.jp/link/ui/2007132341
• Text Lang: en
• Data Source:
  • JaLC
  • NDL
  • Crossref
  • CiNii Articles
• Abstract License Flag: Disallowed