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Hurmadan Izole Edilen Mayaların Bazı Probiyotik

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Hurmadan Izole Edilen Mayaların Bazı Probiyotik HURMADAN İZOLE EDİLEN MAYALARIN BAZI PROBİYOTİK ÖZELLİKLERİNİN ARAŞTIRILMASI Jaafar Nozad Aakef AAKEF YÜKSEK LİSANS TEZİ BİYOLOJİ ANABİLİM DALI GAZİ ÜNİVERSİTESİ FEN BİLİMLERİ ENSTİTÜSÜ NİSAN 2018 Jaafar Nozad Aakef AAKEF tarafından hazırlanan “HURMADAN İZOLE EDİLEN MAYALARIN BAZI PROBİYOTİK ÖZELLİKLERİNİN ARAŞTIRILMASI” adlı tez çalışması aşağıdaki jüri tarafından OY BİRLİĞİ ile Gazi Üniversitesi Biyoloji Anabilim Dalında YÜKSEK LİSANS TEZİ olarak kabul edilmiştir. Danışman: Prof. Dr. Yavuz BEYATLI Biyoloji Anabilim Dalı, Gazi Üniversitesi ...………………… Bu tezin, kapsam ve kalite olarak Yüksek Lisans Tezi olduğunu onaylıyorum. Başkan: Prof. Dr. Gönül DÖNMEZ Biyoloji Anabilim Dalı, Ankara Üniversitesi …………………... Bu tezin, kapsam ve kalite olarak Yüksek Lisans Tezi olduğunu onaylıyorum. Üye: Prof. Dr. Zehra YÜKSEKDAĞ Biyoloji Anabilim Dalı, Gazi Üniversitesi Bu tezin, kapsam ve kalite olarak Yüksek Lisans Tezi olduğunu onaylıyorum. …………………... Tez Savunma Tarihi: 27/04/2048 Jüri tarafından kabul edilen bu tezin Yüksek Lisans Tezi olması için gerekli şartları yerine getirdiğini onaylıyorum. …………………….……. Prof. Dr. Sena YAŞYERLİ Fen Bilimleri Enstitüsü Müdürü ETİK BEYAN Gazi Üniversitesi Fen Bilimleri Enstitüsü Tez Yazım Kurallarına uygun olarak hazırladığım bu tez çalışmasında; Tez içinde sunduğum verileri, bilgileri ve dokümanları akademik ve etik kurallar çerçevesinde elde ettiğimi, Tüm bilgi, belge, değerlendirme ve sonuçları bilimsel etik ve ahlak kurallarına uygun olarak sunduğumu, Tez çalışmasında yararlandığım eserlerin tümüne uygun atıfta bulunarak kaynak gösterdiğimi, Kullanılan verilerde herhangi bir değişiklik yapmadığımı, Bu tezde sunduğum çalışmanın özgün olduğunu, bildirir, aksi bir durumda aleyhime doğabilecek tüm hak kayıplarını kabullendiğimi beyan ederim. Jaafar Nozad Aakef AAKEF 27/04/2018 iv HURMADAN İZOLE EDİLEN MAYALARIN BAZI PROBİYOTİK ÖZELLİKLERİNİN ARAŞTIRILMASI (Yüksek Lisans Tezi) Jaafar Nozad AakefA AKEF GAZİ ÜNİVERSİTESİ FEN BİLİMLERİ ENSTİTÜSÜ Nisan 2018 ÖZET Araştırmada, Libya, Irak ve Türkiye’den satışa sunulan toplam 16 adet hurma çeşidi kullanılmıştır. Bütün hurma örneklerinde mayalara rastlanmış ve sayıları 26,50 - 89,17 log kob/g arasında bulunmuştur. Hurma örneklerinden toplam 16 adet maya suşu izole edilmiştir. Bu suşların koloni ve morfolojik incelemeleri yapıldıktan sonra, moleküler tanımlamaları 18S rRNA dizi analizi ile gerçekleştirilmiştir. Analizler, RefGen Gen Araştırma ve Biyoteknoloji Şirketinde yaptırılmıştır. Tanımlama sonuçlarına göre, bu suşların 14 adedinin Pichia kudriavzeviive 2 adedinin Kluyveromyces marxianusolduğu belirlenmiştir. Tanımlanan türlerin birincil probiyotik seleksiyonlarında yer alan mide asit dirençliliği (pH 2) ve (%0,3) ince bağırsak safra toleranslılıkları belirlenmiştir. Maya suşlarının pH 2 ve %0,3 de inhibisiyonları sırası ile; (%33,41- 96,78) ve (%37,16 -%97,02)) arasında bulunmuştur. Tüm suşların ekzopolisakarit (EPS) üretimleri (108,53 - 45,2 g/L) arasında bulunmuştur. Bu suşların diğer probiyotik özelliklerden olan: agregasyon (otoagregasyon ve koagregasyon), hidrofibosite, antimikrobiyal aktivite ve antibiyotik duyarlılıkları tespit edilmiştir. Tüm suşların otoagregasyon değerleri (%12,47 - 70,96) arasında bulunmuştur. Bu suşların koagregasyon yetenekleri, probiyotik Lactobacillus acidophilus ATCC 4356ve patojen Escherichia coli ATCC 25922 ve Candida albicans ATCC 90028 test mikroorganizmaları ile tespit edilmiş ve sırası ile; (%23,42 - 48,09), (%17,31-%53,38) ve (%16,43- 54,06) arasında bulunmuştur. Suşların hidrofobisite özellikleri, nötral çözücü olan p-kesilen, asidik çözücü olan kloroform ve bazik çözücü olan etil asetatlı ortamlarında incelenmiştir. Tüm suşların bu ortamlarda yüzde hirofobisite değerleri, sırası ile; (%55,78-86,74), (%81,42-84,24) ve (%52,50- 72,23) arasında bulunmuştur. Bu suşların antimikrobiyal etkileri, L. acidophilus ATCC 4356, C. albicans ATCC 10239 ve Pseudomonas auroginosa ATCC, test mikroorganizmaları üzerinde araştırılmıştır. Suşların hiç birinin C. albicans üzerinde antimikrobiyal aktivitesinin olmadığı gözlenirken, L. acidophilus ve P. auroginosa bakterileri üzerinde inhibisyonlarının sırası ile; 8,01- 19,23 mm ve 11,01-24,17 mm zon çapında inhibisiyonları olduğu tespit edilmiştir. Suşların çoğunun test edilen Amoxicillin, Ampicilin, Gentamisin, Tetrasiklin,Trimethoprim antibiyotiklere karşı dirençli oldukları belirlenirken, bazı suşların bazı antibiyotiklere karşı duyarlı oldukları gözlemlenmiştir.Araştırmada, P. kudriavzeii ve K.marxianus suşlarının probiyotik özelliklere sahip oldukları, özelliklerinin cins, tür ve suşlar arasında farklılık gösterdiği gözlemlenmiştir. Bu suşlaradn, P. Kudriavzeii JD1 ve JD2 suşlarının diğer suşlara kıyasla daha üsütün probiyotik özelliklere sahip oldukları belirlenmiştir. Bilim Kodu : 20309 Anahtar Kelimeler : Hurma, Maya, Probiyotik özellikler Sayfa Adedi : 101 Danışman : Prof. Dr. Yavuz BEYATLI v INVESTIGATION OF SOME PROBIOTIC PROPERTIES OF YEASTS ISOLATED FROM DATES (M. Sc. Thesis) Jaafar Nozad Aakef AAKEF GAZİ UNIVERSITY GRADUATE SCHOOL OF NATURAL AND APPLIED SCIENCES April 2018 ABSTRACT In the present study, a total (16) different date samples obtained from Libya, Iraq and Turkey were used. All date samples were presentes yeasts ranging between (26-50 × 89,17 log kob/g). A total, (16) yeast strains were isolated and identified by molecular 18S rRNA sequence analysis. The analyzes were carried out at the RefGen Gen Research and Biotechnology Company. Molecular identification results showed that (14) strains were Pichia kudriavzeii and (2) strains were Kluyveromyces marxianus. The primary probiotic selections of the strains were tested according to the gastric acid (pH 2) and intestinal (%0,3) bile salts tolerances. The inhibition rates of the strains in pH 2 and %0,3 bile saltes ranged between (%33,41-96,78) and (%37,16 -%97,02) respectively. All the strains were produced Exopolysaccharide (EPS) ranging between (108.53 - 452.2 g/L). The other secondary probiotic properties of the strains were tested according to activities related with (aggregation (otoagregation, coagregation), hydrophobicity, antimicrobial activity and antibiotic susceptibilities). The otoaggregation of the strains were ranged between (%12.47 - 70.96). The coaggregation of the strains with probiotic L. acidophilus ATCC 4356, pathogenic E. coli 25922 and C. albicans 90028 were tested and estimated with ranging between (%23,42 - 48,09), (%17,31- %53,38) and (%16,43-54,06) respectively. The hydrophobic activities of the strains were determined in neutral solvent p-ksilen, acidic solvent chloroform and basic solvent ethyl acetate. The hidrophobosity of the strains were estimated and ranged between (%55.78-86.74), (%81.42- 84.24) and (%52.50-72.23) respectively. The antimicrobial activites of the strains on L. acidophilus ATCC 4356, C. albicans ATCC 10239 vand P. auroginosa ATCC were tested. The antimicrobial activites of the strains on L. acidophilus and P. auroginosa were ranged between (8,01-19,23 mm) and (11,01-24,17 mm) inhibition zones. None of the of the strains had showed any antimicrobial activities on C. albicans. Sensitivities of the strains to antibiotics (Amoxicillin, Ampisillin, Gentamycin, Tetracyclin, Trimethoprim) were determined. Generally, all strains were found to be resistant to all antibiotics while few of them were susceptible to some of antibiotics. In this study, all P. kudriavzeii and K.marxianus strains has showed probiotic properties and the characteristics were different between genus, species and strains. P. kudriavzeii JD1 and JD2 strains were found to have highest probiotic properties than the other strains. Science Code : 20309 KeyWords : Dates, Yeast, Probiotic Properties Page Number : 101 Supervisor : Prof. Dr. Yavuz BEYATLI vi TEŞEKKÜR Bu çalışmanın gerçekleştirilmesinde, değerli bilgilerini benimle paylaşan, güler yüzünü ve samimiyetini benden esirgemeyen, her sorun yaşadığımda yanına çekinmeden gidebildiğim ve kullandığı her kelimenin hayatıma kattığı önemini asla unutmayacağım saygıdeğer danışman hocam Sayın Prof. Dr. Yavuz BEYATLI’ya teşekkürü bir borç biliyor ve şükranlarımı sunuyorum. Bu günlere gelmemde büyük öneme sahip olan, desteklerini benden hiçbir zaman esirgemeyen babam ve annem ve kardeşlerime teşekkür ederim, her zaman bana yardımcı olan arkadaşlarım Ümmügülsüm TÜKENMEZ, Kübra ÇELIK, Maihebubai ABUDUREYIMU, Selin UYSAL teşekkür ederim. vii İÇİNDEKİLER Sayfa ÖZET .............................................................................................................................. iv ABSTRACT .................................................................................................................... v TEŞEKKÜR .................................................................................................................... vi İÇİNDEKİLER ............................................................................................................... vii ÇİZELGELERİN LİSTESİ ............................................................................................. x ŞEKİLLERİN LİSTESİ .................................................................................................. xii RESİMLERİN LİSTESİ ................................................................................................. xiii SİMGELER VE KISALTMALAR................................................................................. xiv 1. GİRİŞ .......................................................................................................................
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