Muhammad Bahi ___________________________________________________ Bandamycin as New Antifungal Agent and further Secondary Metabolites from Terrestrial and Marine Microorganisms H N O 2 O O CH HO 3 O NH HO OH OH OH OH O NH2 OH OH O CH3 OH OH OH O OH Cl OH O O CH3 Dissertation Bandamycin as New Antifungal Agent and further Secondary Metabolites from Terrestrial and Marine Microorganisms Dissertation zur Erlangung des Doktorgrades der Mathematisch-Naturwissenschaftlichen Fakultäten der Georg-August-Universität zu Göttingen vorgelegt von Muhammad Bahi aus Banda Aceh (Indonesien) Göttingen 2012 D7 Referent: Prof. Dr. H. Laatsch Korreferent: Prof. Dr. A. Zeeck Tag der mündlichen Prüfung: 17.04.2012 Die vorliegende Arbeit wurde in der Zeit von März 2007 bis November 2011 im Institut für Organische Chemie der Georg-August-Universität zu Göttingen unter der Leitung von Herrn Prof. Dr. H. Laatsch angefertigt. Herrn Prof. Dr. H. Laatsch danke ich für die Möglichkeit zur Durchführung dieser Ar- beit sowie die ständige Bereitschaft, auftretende Probleme zu diskutieren. For my parents, my wife (Anizar) and my sons (Fiqhan, Amirul, Syauqal) Content I Table of Content 1 Introduction .................................................................................................... 1 1.1 Natural products in modern therapeutic use .............................................. 1 1.2 Secondary metabolites from bacteria ......................................................... 5 1.3 Natural products from fungi ..................................................................... 19 2 Scope of the present work............................................................................ 28 3 General techniques ....................................................................................... 30 3.1 Collection of strains ................................................................................. 30 3.2 Strategic procedure in evaluating the selected strains ............................. 30 3.3 Pre-screening ............................................................................................ 32 3.4 Biological screening ................................................................................. 33 3.5 Chemical screening .................................................................................. 34 3.6 Cultivation and scaling-up ....................................................................... 34 3.7 Isolation and purification work-up ........................................................... 35 3.8 Dereplication technique ........................................................................... 35 4 Investigation of selected bacterial strains .................................................. 36 4.1 Marine Streptomyces sp. B8289 .............................................................. 36 4.1.1 2-[5-(2-Oxo-propyl)-tetrahydrofuran-2-yl]-propionic acid ..................... 37 4.1.2 2-[5-(2-Oxo-propyl)-tetrahydrofuran-2-yl]propionic acid methyl ester .. 39 4.1.3 Homononactic acid .................................................................................. 42 4.1.4 Homononactic acid methyl ester .............................................................. 44 4.2 Terrestrial Streptomyces sp. Ank86 ......................................................... 46 4.2.1 4-Hydroxy-2-methoxyacetanilide ............................................................ 47 4.2.2 Actinomycin D ......................................................................................... 48 4.3 Terrestrial Streptomyces sp. Ank68 ......................................................... 51 4.3.1 Fungichromin ........................................................................................... 52 4.4 Terrestrial Streptomyces sp. Ank248 ....................................................... 54 4.4.1 1-Acetyl--carboline ................................................................................ 54 4.5 Terrestrial Streptomyces sp. Ank181 ....................................................... 56 4.5.1 Reductiomycin ......................................................................................... 57 4.6 Ruminal bacterium Enterobacter amnigenus ZIA ................................... 59 4.6.1 Poly-(hydroxybutyric acid) (PHB) .......................................................... 60 II Content 4.6.2 Butyl glycoside ........................................................................................ 61 4.7 Terrestrial Streptomyces sp. Ank75 ......................................................... 63 4.7.1 Benadrostin .............................................................................................. 64 4.7.2 2-O-Methylpyrogallol .............................................................................. 66 4.7.3 N-Aminocarbonyl-2,3-dihydroxybenzamide ........................................... 67 4.7.4 2,3-Dihydroxybenzamide ......................................................................... 69 4.7.5 3-[(1Z)-1-Hexenyl-ONN-azoxyl]-1,2-butanediol .................................... 71 4.7.6 Bandamycin.............................................................................................. 72 4.7.7 N6, N6-Dimethyl-adenosine...................................................................... 77 4.8 Marine Streptomyces sp. B5798 ............................................................... 78 4.8.1 p-Hydroxyphenylacetic acid .................................................................... 79 4.8.2 Macrolactin A........................................................................................... 80 4.9 Terrestrial Streptomyces sp. GT-2005/049 .............................................. 82 4.9.1 Lumichrome ............................................................................................. 83 4.10 Terrestrial Streptomyces sp. GW 4723 .................................................... 86 4.10.1 Celastramycin B ....................................................................................... 86 5 Metabolites from selected endophytic fungi .............................................. 91 5.1 Endophytic fungus LAF20 ....................................................................... 91 5.1.1 Isosclerone................................................................................................ 92 5.1.2 4, 6, 8-Trihydroxy-1-tetralone ................................................................. 94 5.2 Endophytic fungus NP32-A ..................................................................... 96 5.2.1 Cerevisterol .............................................................................................. 96 5.2.2 Fusaproliferin ........................................................................................... 98 5.2.3 Beauvericin ............................................................................................ 100 5.2.4 Trichosetin.............................................................................................. 103 5.2.5 Cerebroside B ......................................................................................... 105 5.3 Endophytic fungus FT44 ........................................................................ 107 5.3.1 1233-A ................................................................................................... 108 5.3.2 Ergosterol peroxide ................................................................................ 109 5.4 Endophytic fungus FTM1 ...................................................................... 112 5.4.1 8-Hydroxy-6,7-dimethoxy-3-methylisocoumarin .................................. 113 5.4.2 5-Methylochracin ................................................................................... 115 Content III 5.4.3 Alternariol 5-methyl ether (AME) ......................................................... 117 6 Plant metabolites ........................................................................................ 119 6.1 Adansonia digitata ................................................................................. 120 6.1.1 Garcinon D ............................................................................................. 120 6.1.2 Rubraxanthone ....................................................................................... 123 6.1.3 Acetylaleuritolic acid ............................................................................. 125 6.2 Dillenia pentagyna ................................................................................. 127 6.2.1 Betulinic acid ......................................................................................... 127 6.2.2 Betulin .................................................................................................... 129 6.3 Kaempferia parviflora ........................................................................... 131 6.3.1 5,7-Dimethoxyapigenin ......................................................................... 131 6.3.2 Ganschisandrine ..................................................................................... 133 6.3.3 Stigmasterol glucoside ........................................................................... 135 6.3.4 Oleanolic acid ........................................................................................ 137 6.3.5 2-Methylanthraquinone .......................................................................... 139 6.3.6 Scopoletin .............................................................................................. 140