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1993 1950 1970 1993 2001 2005 25 (Chang, 2005) 1980 1987 1990 1999 2000 2001 1. 1971 Sasaki (G. applanatum) (Sasaki et al, 1971) -2(IL-2) T (Lai and Lin, 1992) T α DNA T (Lai and Lin, 1991) IL-1,IL-2,IL-6,INF-α,INF-γ (Lieu et al, 1992) 2005 37

(AIDS) β (1-3) β (1-6) G. applanatum G. lucidum G. tsugae (G. applanatum) G. lucidum G. tsugae (Wang et al,1993) T TNF-α IFN-γ (Wang et al, 1997) F3, GLPS, PS-G -1(IL-1) TLR4 (Toll-like receptor-4) (Hsu et al, 2004, Lin et al, 2005) 2-1-2 1982 Kubota G. lucidum A.B. (ganoderic acid A. B.) (Kobota et al,1982) 1999 Kim HIV (Kim and Kim,1999) 1989 Kino G. lucidum LZ- (Ling Zhi-8) LZ-8 110 12,420 Da (Tanaka, 1989) LZ-8 (homodimmer) (systemic anaphylaxis reaction) (Arthus reaction) LZ-8 (Kino, 1989) LZ-8 β-d β-b α-a β-a α-b β-f β-g β-e β-c t-a 1 10 20 30 SDTALIFRLAWDVKKLSFDYTPNWGRGNPN 31 40 50 60 NFIDTVTFPKVLTDKAYTYRVAVSGRNLGV 61 70 80 90 KPSYAVESDGSQKVNFLEYNSGYGIADTNT 91 100 110 IQVFVVDPDTNNDFIIAQWN LZ-8 (Lin, 1997) 38 2005

FIP-gts FIP-fve FIP-vvo (Lin et al. 1997) 1989 Kino LZ-8 LZ-8 (nonobese diabetic, NOD) (Kino, 1990) LZ-8 (immunomodulatory drug) : CsA (cyclosporin A) R FK506 tacrolimus LZ-8 (ven der Hem, 1995) LZ-8 (G. tsugae) 13 kd FIP-gts (fungal immunomodulatory protein-gts) LZ-8 LZ-8 (human peripheral lymphocytes) 3 H-thymidine 5 µg/ml RT-PCR LZ-8 (IL-2, IL-4) (IFN-γ) (TNF-α) 1997 FIP-gts LZ-8 2003 13 kd FIP-fve (Ko et al. 1995) FIP-vvo (Hsu et al. 1997) LZ-8 FIP-fve FIP-vvo 51 2005 G. applanatum G. boninense G. formosamun G. fornicatum G. lucidum G. microsporum G. neojaponicum G. oerstedii G. resinaceum G. sinense G. tropicum G. tsugae G. valesiacum G. weberianum LZ-8 lz-8 LZ-8 genome walking G. microsporum G. fornicatum gmi gfo-1 gfo-2 lz-8 gmi gfo-1 Pichia pastoris KM71 replz regmi regfo-1 MALDI- TOF 2005 39

300 mg/l BALB/c (dendritic cells, DCs) IL-12 J774A.1 TNF-α T Jurkat cells IL-2 regmi 5 µg/ml DCs IL-12 replz 2005 Pichia pastoris (laccase, 1.10.3.2) (manganese peroxidase, 1.11.1.13) (lignin peroxidase, 1.11.1.14) (Servili et al., 2000) (Leonowicz et al., 2001) (Abadulla et al., 2000) (Mougin et al., 2000) S. cerevisiae S. cerevisiae (Larsson et al., 2001) Li Steffens (Li and Steffens, 2002) 1990 Ko (G. lucidum 7071-10) (isozymes) Galc3 ph3-ph10 (Ko et al., 2001) D'souza LZ-8 ( 2005) 40 2005

Veratryl alcohol 0.2U/ml (D'souza et al., 1999) 26 (intron) G. lucidum G. tsugae G. fornicatum RZ.lac4 0814.lac1 1109.lac1 2121 bp 2019 bp 2110bp 9 intron 520 521 521 21 signal peptide Cysteine Phenylalanine AOX1 1109.lac1 Pichia pastoris KM71 ph 3.0 65 C BMMHY 30 C 0.5 662 607 965 AY450404 (Pleurotus ostreatus ) AY176230 (Polyporous ciliatus lcc3) 36146.lac2 933 974 0814.lac1 AY485825 577 1109.lac1 997 998 1109.lac2 947 RZ.lac2 975 999 RZ-2.lac1 RZ-3.lac1 0815.lac1 997 RZ.lac1 RZ-34a.lac1 995 36291.lac1 1.5K 999 36291.lac2 909 0109.lac1 1000 521 615 989 AY364841 (G.sp.BS-1 lac2) 0109.lac2 977 0705.lac1 AF185275 963 0815.lac2 987 RZ.lac3 RZ.lac4 RS.lac1 974 566 864 976 36146.lac1 999 AY414807 (T. pubescens lap2) AY081188 (T. Versicolr lcc3) AY364840 (G.sp.BS-1 lac1) 0109.lac3 997 36291.lac3 1.5K 0705.lac2 0705.lac3 574 36122.lac2 RS.lac3 2005 41

6.6U/ml 81.7U/mg re1109.lac1 55 C 24 100 2005 1970 1995 Global Industry Analysis 2000 27 21 2004 30 2010 60 11.2% 1995 2002 2003 2010 β 2010 (Molecular pharming, biopharming pharming) 80 (Aspergillus spp.) (chymosin) B (virus-like particle) (Burns et al, 2005, Chen et al, 2000) (green fluorescent protein) (Hirano et al, 2000) pan7-1 hygromycin B hph - hygromycin B (1998) 2001 GPD promoter (2001 Sun et al,) 2004 restriction enzyme-mediated integration (2004 Kim et al,) 42 2005

1. 2001 2. 1966 3. 2005 GMI GFO-1 Pichia pastoris 4. 1993. 5. 1990 6. 1998 7. 2005 8. 2003 9. Abadulla E, Tzanov T, Costa S, Robra K.H., Cavaco-Paulo A, Gubitz G.M. 2000. Decolorization and detoxification of textile dyes with a laccase from Trametes hirsuta. Appl Environ Microbiol 66(8):3357-62. 10. Burns, C., K. E. Gregory, M. Kirby, M. K. Cheung, M. Riquelme, T. J. Elliott, M. P. Challen, A. Bailey, Foster G. D. 2005. Efficient GFP expression in the mushrooms Agaricus bisporus and Coprinus cinereus requires introns. Fungal Genet Biol 42:191-9. 11. Chang S.T. 2005. Ganoderma lucidum A prominent source for the healthcare market in the 21th century. Proceedings of the first Symposium on development of China's medicinal fungi industry. 15-27. 12. Chen, X., M. Stone, C. Schlagnhaufer, Romaine C. P. 2000. A fruiting body tissue method for efficient Agrobacterium-mediated transformation of Agaricus bisporus. Appl Environ Microbiol 66:4510-3. 13. D'Souza T.M., Merritt C.S., Reddy C.A. 1999. Lignin-modifying enzymes of the white rot basidiomycete Ganoderma lucidum. Appl Environ Microbiol 65(12):5307-13. 14. Frendscho M.H., Kino, Sone T. and Jardieu P. 1993. Ling Zhi-8:A novel T cell mitogen includes cytokine production and upregulation of ICAM-1 expression. Cell immunol.150,101-133. 15. Hirano, T., T. Sato, K. Yaegashi, Enei H. 2000. Efficient transformation of the edible basidiomycete Lentinus edodes with a vector using a glyceraldehyde-3-phosphate dehydrogenase promoter to hygromycin B resistance. Mol Gen Genet 263:1047-52. 16. Hsu H.C., Hsu C.I., Lin R.H., Kao C.L., Lin J.Y.. 1997. Fip-vvo, a new fungal immunomodulatory protein isolated from Volvariella volvacea. Biochem J 323 (Pt 2):557-65. 17. Hsu H.Y., HUA K.F., Lin C.C., Hsu J., Wong C.H. 2004. Extract of Reishi polysaccharides induces cytokine expression via TLR4-modulated protein kinase signaling pathways. J Immunol. 173(10): 5989-5999. 18. Janusz M.J., Austen K.F. and Czop J.K 1989. Isolation of a yeast heptaglucoside that inhibets monocyte phagocytosis of zymosan particles. J. Immunol. 142: 959-965. 19. Kim S., Song J., Choi H.T. 2004. Genetic transformation and mutant isolation in Ganoderma lucidum by restriction enzymemediated integration. FEMS Microbiology letters 233: 201-204. 20. Kim H.W. and Kim B. K 1999. Biomedicinal triterpenoids of Ganoderma lucidum (Curt.:Fr.)P Karst. (Aphyllophoromycetideae). Intl. J. Med. Mushrooms 1:121-138. 21. Ko E.M., Leem Y.E., Choi H.T. 2001. Purification and characterization of laccase isozymes from the white-rot basidiomycete Ganoderma lucidum. Appl Microbiol Biotechnol 57(1-2):98-102. 2005 43

22. Kino K., Yamsshita A., Yamaoka K., Watanabe J., Tanaka S., Ko K. and Tsunoo H. 1989. Isolation and characterization of a new immunomodulatory protein, Ling Zhi-8(LZ-8),form Ganiderma lucidum. J.Biol. Chem. 264, 472-478. 23. Kino K., Mizumoto K., Sone T., Yamaoka J., Watanabe A., Yamashita K., Yamaoka K., Ko K., Tsunoo H. 1990. An immunomodulatory protein, Ling Zhi-8, prevents insulitis in non-obese diabetic mice. Diabetologia. 33, 713. 24. Ko J.L., Hsu C.I., Lin R.H., Kao C.L., Lin J.Y. 1995. A new fungal immunomodulatory protein, FIP-fve isolated from the edible mushroom, Flammulina velutipes and its complete amino acid sequence. Eur J Biochem 228(2):244-9. 25. Kubota T., Asaka Y., Miura I. and Mori H. 1982. Structures of ganoderic acids A and B, two new lanostane type bitter triterpenes from Ganoderma lucidum(fr.)karst. Helv Chim Acta, 65, 611-619. 26. Larsson S, Cassland P, Jonsson L.J. 2001. Development of a Saccharomyces cerevisiae strain with enhanced resistance to phenolic fermentation inhibitors in lignocellulose hydrolysates by heterologous expression of laccase. Appl Environ Microbiol 67(3):1163-70 27. Leonowicz A, Cho N.S., Luterek J, Wilkolazka A, Wojtas-Wasilewska M, Matuszewska A, Hofrichter M, Wesenberg D, Rogalski J. 2001. Fungal laccase: properties and activity on lignin. J Basic Microbiol 41(3-4):185-227. 28. Lei L.S. and Lin Z.B. 1992. Effect of Ganoderma polysaccharides on T cell subpopulations and production and production of interleukin2 in mixed lymphocyte response. Acta Pharmaceuica Sinica 27(5): 331-335. 29. Lei L.S. and Lin Z.B. 1991. Effect of Ganoderma polysaccharides on the activity of DNA polymerase in -spleen cells stimulated by alloantigens in mice in vitro. T. Beijing Medical University 23(4): 329-333. 30. Li L, Steffens J.C. 2002. Overexpression of polyphenol oxidase in transgenic tomato plants results in enhanced bacterial disease resistance. Planta 215(2):239-47 31. Lieu C.W., Lee S.S. and Wang S.Y. 1992. The effect of Ganoderma lucidum on induction of differentiation in leulsemic U937 cells Anticancer Research 12:1211-1216. 32. Lin Y.L., Liang Y.C., Lee S.S., Chiang B.L. 2005 Polysaccharide purified from Ganoderma lucidum induced activation and maturation of human monocyte-derived dendritic cells by the NF-kappaB and p38 mitogen-activated protein kinase pathways. J Leukoe Biol.78(2):533-543. 33. Miyasaka N., Inoue H., Sone T. and Jardieu P. 1993 Ling Zhi-8:facilitates cellular interaction through modulation of adhesion molecules. Biochem. Biophys. Res. Commun. 186,358-390. 34. Mizuno T., Sakai T. and Chihara G. 1995. Health foods and medicinal usages of mushrooms. Food Review International 11 (1) 69-81. 35. Mougin C, Boyer F.D., Caminade E, Rama R. 2000. Cleavage of the diketonitrile derivative of the herbicide isoxaflutole by extracellular fungal oxidases. J Agric Food Chem 48(10):4529-34. 36. Servili M, DeStefano G, Piacquadio P, Sciancalepore V. 2000. A novel method for removing phenols from grape must. Am. J. Enol. Vitic. 51:357-361. 37. Sun L., Cai H.Q., Xu W.H., Hu Y.L., Gao Y., Lin Z.P. 2001. Efficient transformation of the medicinal mushroom Ganoderma lucidum. Plant Molecular Biology Reporter 19: 383a-383j. 38. van der Hem L.G., van der Vliet J.A., Bocken C.F., Kino K, Hoitsma A.J., Tax W.J. 1995. Ling Zhi-8: studies of a new immunomodulating agent. Transplantation 60(5):438-43. 39. Wang G., Zhang J., Mizuno T., Zhuang C., Ito H., Mayuzumi H., Okamoto H. and Li J. 1993. Antitumor active polysaccharides from the Chinese mushroom Song Shan Lingzhi, the fruiting body of Ganoderma tsugae. Biosci.Biotech. Biochem. 57 (6):894-900. 40. http://www.americamember.org/usa/agriculture.htm 44 2005