Bioactive Substances from Thai Insect Pathogenic Fungi

Transcription

1 Bioactive Substances from Thai Insect Pathogenic Fungi Masahiko Isaka Kanyawim Kirtikara igel L. ywel-jones Morakot Tanticharoen Yodhathai Thebtaranonth ational Center for Genetic Engineering and Biotechnology (BITEC) ational Science and Technology Development Agency (STDA)

2 Biodiversity in Thailand ca 7-10 % of world records Birds World = 9,000 Thailand = >930 Plants World = 250,000 Thailand = 12,000 Thailand Mammals World = 4,000 Thailand =>250 ther Microbes World = 84,000 Expected o. = 1,600,000 Thailand =???? Expected o. = 160,000? Fungi World = 80,000 Expected o. = 1,500,000 Thailand = 3,000 Expected o. = 150,000?

3 MICRRGAISMS SURCES F: 1. MDER MEDICIES: ATIBITICS 2. PRBITICS 3. BIPESTICIDES 4. (IDUSTRIAL) EZYMES etc.

4 e.g. CP 263,114 Isolated from an unidentified fungus isolated from juniper twigs in Texas : exceedingly difficult to synthesize American Type Culture Collection : ATCC Lowering serum cholesterol Isolation : Spencer, P. et al., rg. Lett. 2001, 3, 1443 Synthesis: K.C icolaou group (Angew. Chem. Int. Ed. 2002, 41, 2678) and others

5 CMMERCIALIZATI CEMISTRY STRUCTURES F BIACTIVE CMPUDS BIACTIVITY SCREEIG FERMETATI CDITIS PYLGEETIC CULTURE CLLECTI TAXMY SPECIME (ADDED) VALUE

6 What is an insect (pathogenic) fungus? Cuticle invasion Infection Mycelium Saprophyte (insect s remain = source of food) Producing metabolites To inhibit insect s immune system To act as antibiotic (against competing microorganisms) To modify insect s behavior

7 Winter worm-summer grass 3000 m above sea level snow in winter igel L. ywel-jones field trip to Bhutan

8 April - August

9 Cordyceps sinensis

10 Cordyceps sinensis on Lepidoptera larvae

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13 ther Products from Insect Pathogenic Fungi

14 Cordyceps dipterigena on housefly

15 Cordyceps martialis on butterfly larvae

16 Stroma on Cicada Colony on PDA Cordyceps sp. BCC 1961 Source of isolation : cicada, Thailand Isolated and identified by : Dr. igel L. ywel-jones

17 Cordyceps Khaoyaiensis BCC 1376 Source of isolation : Lepidoptera Larva, Khao Yai ational Park Isolated and identified by : Dr. igel L. ywel-jones

18 Aschersonia tubulata Petch BCC 1785 Source of isolation : omoptera-scale insect, Thailand Isolated and identified by : Dr. igel L. ywel-jones

19 Cordyceps pseudomilitaris BCC 1620 Source of isolation : Lepidoptera Larva, Thailand Isolated and identified by : Dr. igel L. ywel-jones

20 stroma on lepidoptera-pupa Colony on PDA Paecilomyces tenuipes BCC 1614 (Isaria japonica) Source of isolation : Lepidoptera-pupa, Thailand Isolated and identified by : Dr. igel L. ywel-jones

21 Cordyceps sp. on Coleoptera (elaterid larva) BCC 1681 Collected from Khao Soi Dao Wildlife Sanctuary, Chantaburi Cordyceps sp. on Coleoptera (elaterid larva)

22 From Cordyceps sp. BCC 1681 Cordytropolone 1 Anti-malaria (IC 50 ; µg/ml) 2.2 Anticancer (BC / KB; IC 50 ; µg/ml) 2.2 / 17 Cytotoxicity (Vero; IC 50 ; µg/ml) 11 Seephonkai, P., et al., J. Antibiot., 2001, 54,

23 Cordyceps nipponica on ant lions Cordyceps nipponica BCC 1389, Collected from Khao Yai ational Park, Central Thailand

24 Cordypyridones X 2 Atropisomers Antimalarial IC 50 µg/ml : X = 5 : X = >20 / 7.8 Vero Cell IC 50 µg/ml ot tested / >20 1) MCPBA CDCl 3 6 From Cordyceps nipponica BCC ) K 2 C 3 (s) DMF Isaka, M., et al., J. rg. Chem., 2001, 66, R 7 : R = 8 : R = p-bromobenzoyl

25 Cordyceps pseudomilitaris on a Lepidoptera larva Cordyceps pseudomilitaris BCC 1620 Collected from Sam Lan ational Park

26 From Cordyceps pseudomilitaris BCC 1620 Cordyanhydrides 10 9 Compound anti-malaria Cytotoxicity (IC 50 ; µg/ml) (IC 50 ; µg/ml) 9 >100 > >100 >100 Isaka, M., et al., Tetrahedron Lett., 2000, 41,

27 Bioxanthracenes R 1 11 : R 1 = 12 : R 1 = 13 : R 2 = 14 : R 2 = R 2 Compound anti-malaria Cytotoxicity (IC 50 ; µg/ml) (IC 50 ; µg/ml) > > > Atropisomers Atropisomers R 1 R 2 15 : R 1 =, R 2 = 16 : R 1 =, R 2 = 17 : R 1 = Ac, R 2 = 18 : R 1 = Ac, R 2 = Ac 19 : R 1 = Ac, R 2 = 20 : R 1 =, R 2 = From Cordyceps pseudomilitaris BCC 1620

28 From Cordyceps pseudomilitaris BCC 1620 Bioxanthracenes R 1 21 Atropisomers 22 : R 1 = 23 : R 1 = Compound anti-malaria (IC 50 ; µg/ml) Cytotoxicity Vero; (IC 50 ; µg/ml) > >100 Isaka, M., et al., J. Antibiot., 2001, 54, 36-43

29 Paecilomyces tenuipes on a Lepidoptera pupa Paecilomyces tenuipes BCC 1614 Collected from Khlong akha Wildlife Sanctuary, Ranong

30 Verticillium hemipterigenum Verticillium hemipterigenum BCC 1449 Collected from Khlong akha Wildlife Sanctuary, Ranong

31 From Paecilomyces tenuipes BCC 1614 From Verticillium hemipterigenum BCC 1449 From unidentified fungus BCC 2629 R 6 R 3 R S R 1 S R R R 5 S R 4 R 1, R 2, R 3 = Alkyl R 1, R 2, R 3 = Benzyl R 2 Enniatins Cyclodepsipeptides Beauvericins / Allobeauvaricins

32 From Paecilomyces tenuipes BCC 1614 Beauvericins / Allobeauvericins R 6 R 3 R 1 R 2 R 3 R 4 R 5 R 6 R S S R 2 R R S R 5 R 4 R 1 Anti-malaria IC 50 ; µg/ml Anti-TB IC 50 ; µg/ml beauvericin (24) beauvericin A (25) Et beauvericin B (26) Et beauvericin C (27) Et allobeauvericin A (28) allobeauvericin B (29) allobeauvericin C (30) Et Et Et Et Et Et Et Et Et ilanonta, C. et al., Tetrahedron, 2002, 58,

33 From Verticillium hemipterigenum BCC 1449 Enniatins R 6 R 3 S R 1 R 1 R 2 R 3 R 4 R 5 R 6 enniatin B (31) R S R R 5 R 4 R S R 2 Anti-malaria Anti-TB IC 50 ; µg/ml IC 50 ; µg/ml enniatin B 4 (32) enniatin G (33) enniatin C (34) i-bu i-bu i-bu i-bu i-bu i-bu enniatin (35) enniatin I (36) MK1688 (37) s-bu s-bu s-bu s-bu s-bu s-bu enniatin 1 (38a) enniatin 2 (38b) enniatin 3 (38c) i-bu i-bu i-bu s-bu s-bu s-bu ilanonta, C., et al., Tetrahedron, 2003, 59,

34 From unidentified fungus (Acremonium sp.) BCC 2629 R 2 R 1 R Anti-malaria IC 50 ; µg/ml Enniatins Anti-TB IC 50 ; µg/ml 39 : R 1 = R 2 = 40a : R 1 =, R 2 = 40b : R 1 =, R 2 = 41 : R 1 = R 2 = Vongvilai, P., et al., elv. Chim. Acta, 2004, 87,

35 Manuscript ew Enniatins from the Filamentous Fungus Acremonium sp. BCC 2629 Taridaporn Bunyapaiboonsri, * Pornrapee Vongvilai, attapat Lartpornmatulee and Masahiko Isaka Eight new enniatins, P, Q 1, Q 2, Q 3, R 1, R 2, R 3, and S, together with nine known enniatins, L, M 1, M 2,, B,, I, B 4, and MK 1688, were isolated from the filamentous fungus Acremonium sp. BCC 2629.

36 Verticillium hemipterigenum Verticillium hemipterigenum BCC 1449 Collected from Khlong akha Wildlife Sanctuary, Ranong

37 From Verticillium hemipterigenum BCC 1449 Diketopiperazines S R S S 4 42 : R = 44 : R = 43 Compound Anti-malaria Anti-TB Cytotoxicity IC 50 ; µg/ml IC 50 ; µg/ml vero; IC 50 µg/ml 42 > > not tested not tested ilanonta, C., et al., J. Antibiot., 2003, 56,

38 From Verticillium hemipterigenum BCC 1449 Vertihemiptellides S S R S S : R = 46 : R = Compound Anti-TB Cytotoxicity IC 50 ; µg/ml vero; IC 50 µg/ml Isaka, M., et al., rg. Lett., 2005, 7 (11),

39 irsutella nivea BCC 2594 on leaf hopper Collected from Khao Yai ational Park

40 From irsutella nivea BCC 2594 irsutellones R R 47: R = 50 : R = 48 : R = 51 : R = 49 Compound Anti-TB Cytotoxicity IC 50 ; µg/ml vero; IC 50 µg/ml > > not tested Isaka, M., et al., Tetrahedron Lett., 2005, 61,

41 Acknowledgement Biodiversity Research and Training Program (BRT) [co-sponsored by the Thailand Research Fund (TRF) and ational Center for Genetic Engineering and Biotechnology (BITEC)]

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