Mikroalglerin kitin kaynağı olarak kullanımı
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Dosyalar
Tarih
2022
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Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Kitin, N-asetil-D-glikozamin (GlcNAc) birimlerinin β1-β4 bağlarının bir araya gelmesi sonucu oluşan yapısal bir biyopolimerdir. Kimyasal yapısı selüloza ve en yaygın kullanılan türevi kitosana benzemektedir. Ancak bu üç polimer, ikinci karbonlarında farklı fonksiyonel grupların bulunması sebebiyle birbirlerinden ayrılmaktadır. Kitin, yapısında ikinci karbonuna bağlı asetamid (-NHCOCH3) grubu içermektedir. Geniş bir kullanım alanına sahip olan kitin; jelleşmiş formda da, katı formda da kullanılabilmektedir. Diatomlar; silika yapılı frustül adı verilen hücre duvarları olan tek hücreli mikroalglerdir ve β-kitin formunda kitin ürettiği kanıtlanmış bazı türleri içermektedir Bu alanda en çok çalışma bulunan diatom cinsleri Cyclotella ve Thalassiosira'dır. Bu tez çalışmasında, sentrik bir diatom türü ile çalışılmıştır. Tek hücre çekilen türden yoğun kültür oluşturulmuş ve bu kültürden kitin izolasyonu yapılmıştır. Elde edilen diatom türünün genetik karakterizasyonu, kitinin ise yapısal karakterizasyonu gerçekleştirilmiştir. Hücrelerin ve kitinin taramalı elektron mikroskobu (SEM) ve ters faz kontrast mikroskobu ile görüntüleri alınmıştır. Farklı konsantrasyonlardaki kitinin fibroblast hücreleri üzerindeki sitotoksik etkisi 3-(4,5-dimetiltiyazol-2-yl)-2-5-difeniltetrazolyum bromür (MTT) ve Nötral Kırmızı Alım (NKA) testleri ile yorumlanmıştır. Karakterizasyonlar sonrası; diatom hücresi Thalassiosirales takımına ait bir tür olarak tanımlanmış ve elde edilen malzemenin β formunda kitin olduğu sonucuna varılmıştır. Izole edilen kitin konsantrasyonu yaklaşık olarak 25 mg/L'dir. Sitotoksisite testleri, kitinin 50 μg/ml'ye kadar toksik etki göstermediğini ortaya koymuştur.
Chitin is a structural biopolymer formed as a result of β1-β4 bonds of N-acetyl-D-glucosamine (GlcNAc) units. Its chemical structure is similar to cellulose and its most widely used derivative, chitosan. However, these three polymers differ from each other due to the presence of different functional groups on their second carbon. Chitin contains acetamide (-NHCOCH3) group attached to its second carbon in its structure. The chitin, which has a wide range of uses can be used in gel form or solid form. Diatoms are single-celled microalgae with cell walls called frustules made of silica and contain some species proven to produce chitin in the form of β-chitin. The most studied diatom genus in this field are Cyclotella and Thalassiosira. In this thesis, a centric diatom species was studied. Intensive culture was formed from the single-celled strain, and chitin was isolated from this culture. The genetic characterization of the obtained diatom species and the structural characterization of chitin were performed. Images of cells and chitin were taken with scanning electron microscopy (SEM) and inverted phase contrast microscopy. The effect of chitin on the viability of fibroblast cells at different concentrations was interpreted by cytotoxicity tests 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and neutral red uptake assays. After characterizations; the diatom cell was defined as a species belonging to the order Thalassiosirales and it was concluded that the material obtained was chitin in the β form. The concentration of isolated chitin was approximately 25 mg/L. The cytotoxicity tests have resulted in chitin not showing any toxic effects up to 50 μg/ml.
Chitin is a structural biopolymer formed as a result of β1-β4 bonds of N-acetyl-D-glucosamine (GlcNAc) units. Its chemical structure is similar to cellulose and its most widely used derivative, chitosan. However, these three polymers differ from each other due to the presence of different functional groups on their second carbon. Chitin contains acetamide (-NHCOCH3) group attached to its second carbon in its structure. The chitin, which has a wide range of uses can be used in gel form or solid form. Diatoms are single-celled microalgae with cell walls called frustules made of silica and contain some species proven to produce chitin in the form of β-chitin. The most studied diatom genus in this field are Cyclotella and Thalassiosira. In this thesis, a centric diatom species was studied. Intensive culture was formed from the single-celled strain, and chitin was isolated from this culture. The genetic characterization of the obtained diatom species and the structural characterization of chitin were performed. Images of cells and chitin were taken with scanning electron microscopy (SEM) and inverted phase contrast microscopy. The effect of chitin on the viability of fibroblast cells at different concentrations was interpreted by cytotoxicity tests 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and neutral red uptake assays. After characterizations; the diatom cell was defined as a species belonging to the order Thalassiosirales and it was concluded that the material obtained was chitin in the β form. The concentration of isolated chitin was approximately 25 mg/L. The cytotoxicity tests have resulted in chitin not showing any toxic effects up to 50 μg/ml.
Açıklama
Anahtar Kelimeler
Diatom, kitin, β-kitin, biyoteknoloji, chitin, β-chitin, biotechnology
