Cloning and purification of biosynthetic enzymes
Başlık:
Cloning and purification of biosynthetic enzymes
Yazar:
Gürelme, Ebru, author.
Yazar Ek Girişi:
Fiziksel Tanımlama:
xi, 63 leaves:+ 1 computer laser optical disc.
Özet:
Bioproducts have become prominent with their sustainable, eco-friendly and renewable features. In recent years, research and development studies focused on production of biodiesel and pharmaceuticals. Biodiesel can be synthesized in the form of fatty acid ethyl ester (FAEE) by in vivo activity. This synthesis is catalyzed by wax ester synthases (WS). This study aims cloning and purification of wax ester synthases from Psychrobacter arcticus 273-4 (PaWES) and Mus musculus C57BL/6 (MmWES). PaWES was cloned and expressed by Escherichia coli BL21(DE3) strain, at the proper conditions with using pET expression system. It was purified with approximately 1 mg yield. Cloning of the second wax ester synthase (MmWES) was achieved to Saccha-romyces cerevisiae and it was purified with lower than 1 mg yield. The other aim of this study is related to taxadien-5α-ol-O-acetyltransferase from Taxus cuspidata (TcT5AT). This enzyme belongs to the biosynthesis pathway of Taxol, which is the most commonly used chemotherapy drug. Cloning and purification studies of this enzyme were successfully performed. It was expressed by Escherichia coli BL21(DE3) Star strain and purified with the yield of 23 mg. Immobilized Metal Affinity Chromatog-raphy (IMAC) is used for all three enzymes as a purification strategy. This project can pave the way for structural studies of all biosynthetic enzymes mentioned above. In summary, the findings of this study will circuitously help for solving the relationship between function and structure of these enzymes. It may lead to increased generation of Taxol and FAEE based biodiesel.
Yazar Ek Girişi:
Tüzel Kişi Ek Girişi:
Tek Biçim Eser Adı:
Thesis (Master)--İzmir Institute of Technology: Biotechnology and Bioengineering.
İzmir Institute of Technology: Biotechnology and Bioengineering--Thesis (Master).
Elektronik Erişim:
Access to Electronic Versiyon.