Ege bölgesi kaplıca kaynakları ve çevresinden izole edilen termofilik aktinomisetlerden yeni antimikrobiyal moleküllerin elde edilmesi
Isolation of new antimicrobial molecules from thermophilic actinomycetes in living hot spring sources and environments of aegean region
Yazar: ÇAĞLA AKMEMİŞ KORKMAZ
Danışman: DOÇ. DR. ERDAL BEDİR
Ege Üniversitesi / Fen Bilimleri Enstitüsü / Biyomühendislik Ana Bilim Dalı
Derece: Yüksek Lisans, 2006
Konu: Biyomühendislik = Bioengineering

Özet:
Bu çal mada Ege Bölgesindeki 59 s cak su kayna ndan 63 toprak,50 su ve 28 sediment örne i (toplam 141) toplanm ve 146 termofilikAktinomiset kültürü izole edilmi tir. Bir dizi testler yap larak izolatlar ns cakl k (650C), pH (11) ve tuz (5%) toleranslar belirlenmi tir. Safkültürlerin antibakteriyel aktiviteleri Agar-Plak metodu kullan larakStaphylococcus aureus ATCC 6538 P, Enterecoccus faecalis ATCC 29212,Klebsiella pneumoniae CCM 2318, Escherichia coli ATCC 29998,Escherichia coli O157H7 ve metisilin dirençli Staphylococcus aureus(MRSA) organizmalar na kar test edilmi tir.146 termofilik Aktinomisetten 98 izolat dar ve/veya geni spektrumaral nda antimikrobiyal etki göstermi tir. MRSA ya kar en yüksekaktiviteyi gösteren 66-2 nolu izolat biyoaktivite rehberli izolasyon içinseçilmi tir. Fermentasyon ve devam nda solvan ekstraksiyonu (H2O-Hekzan, H2O-EtOAc, H2O-n-BuOH) yap ld nda en yüksek aktivite EtOAcekstresinde görülmü tür. Biyoaktivite rehberli izolasyon çal malar EtOAcfaz nda kromatografik metotlar (Preparatif .T.K., VSK, FlashKromatografi)) kullan larak gerçekle tirilmi ve sonuçta aktif metabolitsafla t r lamamas na ra men, ikisi yeni ve üçü bilinen olmak üzere besekonder metabolit elde edilmi tir. Yeni bile iklerin yap s 1D- ve 2D-NMRve LC-ESI spektral metodlar kullan larak N-(2-(1H-indol-3-il)etil)-2-(1-aminyumetil)tiazol-4-karboksamid ve N-(2-(1H-indol-3-il)etil)-2-(1-(propiyonamido)etil)tiazol-4-karboksamid olarak belirlenmi tir.Anahtar sözcükler: Termofilik Aktinomiset, fermentasyon, sekondermetabolit, antibiyotikler.

Abstract:
In this study, 63 soil, 50 water and 28 sediment samples were (total of141) collected from 59 hot spring sources of the Aegean Region and 146thermophilic Actinomycetes cultures were isolated. Temperature (650C), pH(11) and saline (5%) tolerance of the isolates were determined by some tests.The antibacterial activities of pure isolates were tested against Staphylococcusaureus ATCC 6538 P, Enterecoccus faecalis ATCC 29212, Klebsiellapneumoniae CCM 2318, Escherichia coli ATCC 29998, Escherichia coliO157H7 and methicillin resistant Staphylococcus aureus (MRSA) by usingagar-plaque method.Ninety eight isolates were found to possess antimicrobial activityshowing from broad to narrow spectrum. Based on the highest antibacterialactivity versus MRSA, the isolate 66-2 was selected for bioassay-guidedisolation. Fermentation followed by solvent partition studies (H2O-Hexane,H2O-EtOAc, H2O-n-BuOH) showed that the highest activity was present inEtOAc extract. Bioassay guided isolation studies performed on EtOAcextract by using chromatographic methods [Preparative TLC, Open ColumnChromatography, VLC, Flash Chromatography], resulted in the isolation oftwo new and three known compounds, but active metabolite. The structuresof the new compounds were elucidated as N-(2-(1H-indol-3-yl)ethyl)-2-(1-aminiumethyl)thiazole-4-karboxamide and N-(2-(1H-indol-3-yl)ethyl)-2-(1-(propionamido)ethyl)thiazole-4-karboxamide by using spectralmethods, 1D and 2D NMR and LC-ESI-MS.Key words: Thermophilic Actinomycetes, fermentation, secondermetabolite, antibiotics.