Daedaleopsis tricolor

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1 ~ Mycosystema Daedaleopsis tricolor HPLC (2- ) ,8-3 2 Q939.5 A The antifungal components from Daedaleopsis tricolor XIANG Yun-Bin 1, 2 LI Er-Wei 2 DING Gang 2 ZHANG Xiao-Qing 2 YU Zhi-He 1* ( 1 College of Life Science, Yangtze University, Jingzhou , China; 2 Institute of Microbiology, Chinese Academy of Sciences, Beijing , China) Abstract: Through screening antifungal components from secondary metabolites of polypores, a strain of Daedaleopsis tricolor was found to produce metabolites that showed antifungal activities against Candida albicans AS2.2086, Saccharomyces cerevisiae AS2.1882, Aspergillus flavus AS and Aspergillus fumigatus AS The solid culture of D. tricolor was extracted with methyl ethyl ketone and the extracts were fractionated by vacum liquid chromatography, Sephadex LH-20 and HPLC. Three isocoumarins (1-3) were identified as 4-(2-hydroxyethanol)-8-hydroxyisocoumarin, 4-(2-hydroxyacetyl)-8-hydroxyisocoumarin and 4-(2-hydroxyacetyl)-5, 8-hydroxyisocoumarin by analyses of their ESIMS and NMR data. These compounds were isolated for the first time from the genus Daedaleopsis and compound 2 showed antifungal activity. Key words: polypores, secondary metabolites, isococumarin, antifungal activity Daedaleopsis tricolor(bull.: Fr.)Boudartsev et Singer Dembitsky et al Rösecke & König 2000 Kim (29)-lupen-3-one * Corresponding author. zhiheyu@hotmail.com

2 % 90% AS Bruker Esquire 3000 plus Bruker Avance-400 Sephadex LH-20 Sigma HPLC Aglient 1100 C 18 25cm 10µm 1.3 PDA 258d 0.4%1%0.4%25 160r/min 7d 10%10150mL 20g 60g 7.5g 7g MgS 4 7H 2 2.5g FeS 4 7H 2 1g ZnS 4 7H 2 10mg CaC 3 2.5g 200g 1000mL 500mL25 35d 1.4 Escherichia coli AS Pseudomonas aeruginosa AS Staphylococcus aureus AS Micrococcus luteus AS Candida albicans AS Saccharomyces cerevisiae AS Aspergillis fumigatus AS Aspergillus flavus AS mueller-hinton agar sabouraud dextrose agar PDA CPU/mL 4 Wicklow et al mL 3 20mg/mL 10µL5mm 371d252d25

3 4 Daedaleopsis tricolor 567 3d 20mg/mL1mg/mL : 100:1 0:100 v/v:100:1 40:60 v/v TLC 20mg/mLSephadex LH-20 : 1:4 v/v:4:1 v/v: 2:3 v/v TLC20mg/mL Sephadex LH-20 HPLC 20mg/mL g 1.5g 7mg HPLC 0.5g Sephadex LH-20fr4-fr8 fr4-fr6fr7 fr8 Sephadex LH-20 HPLC 1 20mg 2 2mg 3 5mg oospoglycol C 11 H 10 5 ESIMSm/z 220.7[M-H] [M+N a ] + 1 H NMR 400M Hz CD 3 CCD 3 δ H m H H ddd H H t H d J=3.88Hz H m H H d J=8.16Hz H H d J=7.92Hz H H s H H dd J=7.92Hz 8.16Hz H H s 8-13 CNMR 400M Hz CD 3 CCD 3 δ66.5 C C C-8a C C C C-4a C C C C-1 13 CNMRS Sonnenbichler et al (2- )-8- oosponol C 11 H 8 5 ESIMSm/z 219[M-H] - 1 H NMR 400M Hz CD 3 CCD 3 δ H brs H s H H d J=8.16Hz H H t J=8.16Hz H H d J=8.16Hz H H s H H s 8-13 CNMR 400M Hz CD 3 CCD 3 δ67.0 C C-8a C C C C-4a C C C C C-9 1 H NMR Nakajima et al ,8-C 11 H 10 6

4 ESIMSm/z 237[M-H] - 1 H NMR 400M Hz CD 3 D δ H dd H H d H H H H d J=8.20Hz H H d J=8.20Hz H H s H-3 H H Compound 1 Compound 2 Compound fr8 Sephadex LH fr4-fr8 fr7 fr8 1~3 1mg/mL 2 1 Table 1 The antibiotic activities of the crude extract, the fractions and the compounds isolated from the fermentation of Daedaleopsis tricolor Antibiotic zone (mm) m fr1 fr2 fr3 fr4 fr5 fr6 fr7 fr8 fr9 fr10 fr Escherichia coli Pseudomonas aeruginosa Staphylococcus aureus Micrococcus luteus Candida albicans Saccharomyces cerevisiae Aspergillus flavus Aspergillus fumigatus m fr1-11 Sephadax LH Note m: crude extract; fr1-11: the fractions isolated by Sephadex LH-20; 1-3: the compounds 1-3.

5 4 Daedaleopsis tricolor Trametes versicolor Trametes robiniophila Phellinus igniarius Taiwanofungus camphoratus Wolfiporia cocos (2- )-8- osponol Trichophyton asteroides 1.56µg/mL 1.56µg/mL 3.12µg/mL Nakajima et al osponol [REFERENCES] Dembitsky VM, Rezanka T, Shubina EE, Unusual hydroxy fatty acids from some higher fungi. Phytochemistry, 34: 1057~1059 Kim EM, Jung HR, Min TJ, Purification, structure determination and biological activities of 20 (29)-lupen-3-one from Daedaleopsis tricolor (Bull. ex Fr.) Bond. et Sing. Bull Korean Chem Soc, 22: 59~62 Mao XL, Macrofungi in China. Henan Science and Technology Press, Henan.1~719 (In Chinese) Nakajima S, Kawai KI, Yamada S, The identification of Lenzitin as oosponol. Phytochemistry, 15: 327 Rösecke J, König WA, Constituents of the fungi Daedalea quercina and Daedaleopsis confragosa var. tricolor. Phytochemistry, 54: 757~762 Sonnenbichler J, Sonnenbichler I, Schwarz D, Biosynthesis of oospoglycol elucidated by 13 CNMR. Phytochemistry, 44: 267~269 Wicklow DT, Joshi BK, Gamble WR, Gloer JB, Dowd PF, Antifungal metabolites (Monorden, Monocillin IV, and Cerebrosides) from Humicola fuscoatra Traaen NRRL 22980, a Mycoparasite of Aspergillus flavus Sclerotia. Appl Environ Microbiol, 64: 4482~ ~719