Cloning and expression of the pseudomonas KE38 extra-cellular lipase gene in E. coli
Karakaş, Fulya.

Cloning and expression of the pseudomonas KE38 extra-cellular lipase gene in E. coli

Karakaş, Fulya.

Yazar Ek Girişi
Karakaş, Fulya.

Yayın Bilgileri
[s.l.]: [s.n.], 2013.

Fiziksel Tanımlama
ix, 33 leaves.: ill. + 1 computer laser optical disc.

Lipases are serine hydrolases that catalyze both the hydrolysis and synthesis of insoluble or poorly soluble long-chain triacylglycerols with an acyl chain length ≥ 10 carbon atoms based on the presence or absence of water. Lipases are produced and secreted by all kingdoms of life that are eukaryotes including plants, animals, fungi and prokaryotes including bacteria and archaea. However, microbial lipases, especially from bacteria, more useful than their plant and animal derivatives because of several important properties. Because of their acitivities in both aqueous and nonaqueous environments, lipases have specific applications in industry and medicine. The primary goals of this thesis were to clone and express the extra-cellular lipase gene from Pseudomonas sp. KE38, isolated from soil samples of Erciyes mountain in Kayseri, in E. coli and partial purification of the gene product. To achieve this aim, genome walking technique was used to obtain lipase gene from Pseudomonas sp. KE38, that gene was then cloned into pET28a expression vector and expressed in E. coli. The lipase expression of E. coli BL21 and its activity was screened with olive oil-Rhodamin B plate assay. After expression recombinant lipase was partially purified via inclusion body isolation. Moreover the optimum lipase production time of E. coli BL21 cells were determined and analyzed with SDS-PAGE. According to SDS-PAGE analysis the recombinant lipase was approximately 64 kDa and lipase production reached to the highest level after two hours of IPTG induction. As conclusion, recombinant lipase from Pseudomonas sp. KE38 was cloned into E. coli, expressed and partially purified.

Konu Başlığı
Lipase -- Biotechnology
Molecular cloning.
Escherichia coli.

Yazar Ek Girişi
Arslanoğlu, Alper.

Tüzel Kişi Ek Girişi
İzmir Institute of Technology. Biotechnology and Bioengineering.

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.

LibraryMateryal TürüDemirbaş NumarasıYer NumarasıDurumu/İade Tarihi
IYTE LibraryTezT001133TP248.65.E59 K18 2013Tez Koleksiyonu