Year 2020, Volume 23 , Issue 5, Pages 1135 - 1143 2020-10-31

Trichophyton sp. Suşundan Keratinaz Üretimi, Saflaştırılması ve Kısmi Karakterizasyonu
Isolation and Partial Characterizatıon of Keratinase from Trichophyton sp.

Dönay PARLAK [1] , Hüseyin TANIŞ [2] , Ashabil AYGAN [3]


Keratinaz aktivitesi gösteren Trichophyton sp. Tr-9 suşu keratin tozu- feather meal içeren minimal besiyerinde büyütürek enzim üretimi gerçekleştirilmiştir. Süpernatanttan Amonyum sülfat çöktürme sonrası, Sephadeks G-100 ve DEAE Sepharose kolon kromatografi uygulamaları ile enzim saflaştırılması yapılmıştır. Maksimum keratinolitik aktivite pH 7.5 ve 37°C’de gözlenmiştir. Enzim pH 5.5-8.0 ve 20°C-40°C sıcaklık aralıklarında stabilite gösterirken, enziminin moleküler ağırlığı SDS page ve zimogram analizlerinde yaklaşık 34 kDa olarak hesaplandı. CaCI2 (5mM) keratinaz aktivitesini (%148) stimüle edici etki gösterdi. Diğer taraftan EDTA (5mM) ve SDS (%1) sırasıyla %49, % 49 etki göstererek kısmen inhibe etti, PMSF (1mM ve 5mM) ile tam inaktivasyon elde edildi. Sonuç olarak, enzimin fiziko-kimyasal özellikleri birçok endüstriyel ve biyoteknolojik uygulamalarda kullanışlı olabileceğini göstermiştir.

Keratinase enzyme production was accomplished from Trichophyton sp. Tr-9 in minmal medium containing feather meal. Enzyme was purified with Ammonium sulfate, Sephadeks G-100 and DEAE Sepharose column. Maxiumum keratinolitik activity was obtained at pH 7.5 and 37°C. Enzyme was higly stabil between pH 5.5-7.5 and 20-40oC. With the SDS-Page analysis of the enzyme, molecular weight of the enzyme was calculated as 34 kDa. CaCI2 (5mM) had a stimultory effect (148%) on enzyme. On the other hand, EDTA (5mM) and SDS (%1) inhibited enzyme acitivity up to 49% and 49%, respectively. PMSF (1- 5mM) has strongly inhibited enzyme. As a result, physico-chemical properties of the enzymeshowed that Trichophyton sp. Tr-9 could be useful in various industrial and biotechnological applications.

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Primary Language tr
Subjects Biology
Journal Section RESEARCH ARTICLE
Authors

Orcid: 0000-0002-8940-5842
Author: Dönay PARLAK
Institution: Kahramanmaras Sütcü Imam University
Country: Turkey


Orcid: 0000-0002-2012-7864
Author: Hüseyin TANIŞ (Primary Author)
Institution: KSÜ Fen Edebiyat Fakültesi
Country: Turkey


Orcid: 0000-0003-4936-9872
Author: Ashabil AYGAN
Institution: KAHRAMANMARAŞ SÜTÇÜ İMAM ÜNİVERSİTESİ
Country: Turkey


Supporting Institution K.S.Ü. BAP
Project Number 2009/6-4M
Thanks Bu çalışma KSU Bilimsel araştırmalar birimi tarafından 2009/4-6M Nolu proje ile desteklenmiştir.
Dates

Application Date : January 22, 2020
Acceptance Date : March 13, 2020
Publication Date : October 31, 2020

APA Parlak, D , Tanış, H , Aygan, A . (2020). Trichophyton sp. Suşundan Keratinaz Üretimi, Saflaştırılması ve Kısmi Karakterizasyonu . Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi , 23 (5) , 1135-1143 . DOI: 10.18016/ksutarimdoga.vi.678756