Saccharomyces cerevisiae kaynaklı invertaz enzimi üretimine ultrasonikasyon parametrelerinin etkisinin araştırılması
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Tarih
2021
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Tekirdağ Namık Kemal Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
İnvertaz enzimi bilimsel dilde ß-fruktofuranozidaz ya da sükraz olarak bilinmektedir. Hidrolaz sınıfına ait olan bu enzim kaynak olarak hayvan, bitki ya da mikroorganizmalardan yararlanılarak elde edilebilmektedir. İnvertaz enzimi intrasellüler yani hücre içerisinde faaliyet gösteren bir yapıya sahiptir. Protein yapısında olan invertaz enzimi herhangi bir dış faktör olmadıkça tüm faaliyetini hücre duvarından geçemediği için hücre içerisinde gerçekleştirir. Bu çalışmada Saccharomyces cerevisia kaynaklı ekmek mayası fabrikası atığı olan eleküstü mayadan invertaz üretimi gerçekleştirilmiştir. S. cerevisia'dan intraellüler yapıda olan invertaz enzimini üretebilmek için ultrasonikasyon cihazı kullanılarak ses dalgaları yardımı ile hücre yapısına zarar verilmiştir. Elde edilen enzimin protein miktarı ve enzim aktivitesinin maksimum düzeyde olabilmesi için ultrasonikasyon cihazı için süre, güç (amplitüt) ve maya miktarı parametreleri gerçekleştirilen deneylerde araştırılmıştır. Cevap yüzey yöntemi kullanılarak rastgele oluşturulan 20 farklı deneme modeli elde edilmiştir. Gerçkeleştirilen deneyler sonucunda en yüksek protein miktarının 23,92 mg/ml'de, 45 dakika, %70 amplitüt ve % 36,82 gram değerlerinde olduğu, enzim aktivitesinin ise en yüksek olduğu değerlerin 1720 IU/ml, 45 dakika, % 70 amplitüt ve % 36,82 gram olduğu tespit edilmiştir. Çalışmadaki amaç en yüksek protein miktarı ve invertaz enzimi aktivitesine ulaşmaktır. Bu sebeple kullanılan parametre aralıklarında belirlenen model denklemin tahmin başarısını ölçmede daha anlamlı sonuçlar vereceğinden kullanılan paket programda "response optimizer" seçeneği ile en yüksek değerleri sağlayan parametreler belirlenmiştir. Protein tayini optimizasyonu için % 49,5, enzim aktivitesi için %52 başarı sağlanabilmiştir.
Invertase enzyme is scientifically known as ß-fructofuranosidase or sucrase. This enzyme, which belongs to the hydrolase class, can be produced by animals, plants, or microorganisms. Invertase is an intracellular enzyme and it cannot pass through the cell wall. Invertase was produced by a waste product including sieve yeast (Saccharomyces cerevisiae), obtained from a baker yeast factory. In order to extract the invertase enzyme, the cell was disrupted by an ultrasonication device using sound waves. Since the protein amount and enzyme activity of the obtained enzyme should be at maximum level, the parameters of time, power (amplitude) of ultrasonication, and yeast amount were investigated in the experiments. In the study, 20 different experiments were conducted which were randomly generated using the response surface methodology. As a result of the experiments, the highest protein amount was found as 23.92 mg protein/ml crude extract using following parameters: 45 min ultrasonication time, 70% amplitude, and 36.82% yeast concentration. Also, the highest enzyme activity was found to be 1720 IU/ml crude extract using following parameters: 45 min ultrasonication time, 70% amplitude, and 36.82% yeast concentration. The aim of the study was to reach the highest protein amount and invertase enzyme activity. The model equations were used to predict the amount of protein and invertase activity in crude extract at optimum conditions: ultrasonication time, amplitude and yeast concentration. Finally, it was recorded that the accuracy of the model for prediction of protein amount was 49.5% and that of invertase activity was 52%.
Invertase enzyme is scientifically known as ß-fructofuranosidase or sucrase. This enzyme, which belongs to the hydrolase class, can be produced by animals, plants, or microorganisms. Invertase is an intracellular enzyme and it cannot pass through the cell wall. Invertase was produced by a waste product including sieve yeast (Saccharomyces cerevisiae), obtained from a baker yeast factory. In order to extract the invertase enzyme, the cell was disrupted by an ultrasonication device using sound waves. Since the protein amount and enzyme activity of the obtained enzyme should be at maximum level, the parameters of time, power (amplitude) of ultrasonication, and yeast amount were investigated in the experiments. In the study, 20 different experiments were conducted which were randomly generated using the response surface methodology. As a result of the experiments, the highest protein amount was found as 23.92 mg protein/ml crude extract using following parameters: 45 min ultrasonication time, 70% amplitude, and 36.82% yeast concentration. Also, the highest enzyme activity was found to be 1720 IU/ml crude extract using following parameters: 45 min ultrasonication time, 70% amplitude, and 36.82% yeast concentration. The aim of the study was to reach the highest protein amount and invertase enzyme activity. The model equations were used to predict the amount of protein and invertase activity in crude extract at optimum conditions: ultrasonication time, amplitude and yeast concentration. Finally, it was recorded that the accuracy of the model for prediction of protein amount was 49.5% and that of invertase activity was 52%.
Açıklama
Anahtar Kelimeler
İnvertaz Enzimi, Ultrasonikasyon, Optimizasyon, Invertase Enzyme, Ultrasonication, Optimization, Biyoteknoloji, Biotechnology
