Expression, purification, and characterization of bovine chymosin enzyme using an inducible pTOLT system      
Yazarlar (4)
Yakup Ulusu
Karamanoğlu Mehmetbey Üniversitesi, Türkiye
Doç. Dr. Sema BİLGİN Tokat Gaziosmanpaşa Üniversitesi, Türkiye
Hülya Kuduğ
Tokat Gaziosmanpaşa Üniversitesi, Türkiye
Prof. Dr. İsa GÖKÇE Tokat Gaziosmanpaşa Üniversitesi, Türkiye
Makale Türü Açık Erişim Özgün Makale
Makale Alt Türü SSCI, AHCI, SCI, SCI-Exp dergilerinde yayınlanan tam makale
Dergi Adı PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
Dergi ISSN 1082-6068 Wos Dergi Scopus Dergi
Dergi Tarandığı Indeksler SCI-Expanded
Dergi Grubu Q4
Makale Dili İngilizce
Basım Tarihi 01-2016
Cilt No 46
Sayı 6
Sayfalar 596 / 601
DOI Numarası 10.1080/10826068.2015.1085399
Makale Linki http://dx.doi.org/10.1080/10826068.2015.1085399
Özet
In recent years, various studies in the field of industrial enzymes of biotechnology have gained importance due to increasing development in enzyme technology. The different areas where enzymes are used and their economic value of biotechnological products further increases their importance. There are hundreds of different types of cheese but each is made by coagulating milk using rennet to give curds. Today, researchers have begun to develop alternative systems in the cheese industry related to milk-clotting enzymes. In this study, the nucleic acid sequence encoding the optimized chymosin enzyme was used and cloned by Not I and Mlu I restriction enzymes into pTOLT vector system. Then using this construct, the enzyme as a fusion with Tol-A-III protein was produced in Escherichia coli BL21 (DE3) cells. After disrupting the E. coli cell and separating from the constituents by high speed centrifugation, the enzyme was purified by affinity chromatography and fractions were analyzed by SDS-PAGE. Purified enzyme has shown its activity. Optimum temperature and pH of CHY-Tol-A-III protein were 40°C and 6.5, respectively.
Anahtar Kelimeler
Chymosin | Escherichia coli | inducible pTOLT system | recombinant protein production | rennet | TOL-A-III