Cloning, heterologous expression and purification of various wax ester synthases in Escherichia coli
Başlık:
Cloning, heterologous expression and purification of various wax ester synthases in Escherichia coli
Yazar:
Ovacık, Kamil, author.
Yazar Ek Girişi:
Fiziksel Tanımlama:
xii, 45 leaves: illustrarions, charts;+ 1 computer laser optical disc.
Özet:
Biodiesel, known all around theWorld, is a diesel fuel containing fatty acid methyl esters (FAMEs) and fatty acid ethyl esters (FAEEs) with different molecular weights. The recent studies which are about the development of FAEE focused on production of FAEEs in vivo syntheses. This synthesis is catalyzed by wax ester synthases (WS). Bifunctional wax ester synthase/acyl-coenzyme-A (acyl-CoA): diacylglycerol acyltransferase (WS/DGAT) synthesizes wax ester by processing a certain range of fatty alcohols and fatty acyl-CoAs. It is considered as the final enzyme in biosynthetic process of wax ester production. Aim of the research is cloning, heterologous expression, purification and crystallization trial of was ester synthases from M. aquaeolei VT8 (MaWES) and R. opacus PD630 (RoWES). MaWES was cloned into pET expression vector and heterologous expression of MaWES was carried out in E.coli BL21 (DE3) strain. Three chromatography systems were used for purification of MaWES. After Immobilized Metal Affinity Chromatography (IMAC), buffer exchange and gel filtration chromatography, enzyme was purified with approximately 100 mg yield. This project can pave the way for structural studies WS/DGAT 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 FAEE based biodiesel.
Yazar Ek Girişi:
Tüzel Kişi Ek Girişi:
Tek Biçim Eser Adı:
Thesis (Master)--İzmir Institute of Technology: Molecular Biology and Genetics.
İzmir Institute of Technology: Molecular Biology and Genetics.--Thesis (Master).
Elektronik Erişim:
Access to Electronic Versiyon.