Functional expression of an alkaline lipase inEscherichia coli

Autor:	Zahra Etemadifar, Koorosh Goodarzvand Chegini, Mohsen Ani, Mohammed Rabbani, Hamid Mirmohammad-Sadeghi, Fatemeh Moazen
Rok vydání:	2009
Předmět:	chemistry.chemical_classification biology Triacylglycerol lipase Substrate (chemistry) medicine.disease_cause Applied Microbiology and Biotechnology Enzyme assay law.invention Enzyme Biochemistry chemistry law biology.protein medicine Recombinant DNA Lipase Escherichia coli Thermostability
Zdroj:	Annals of Microbiology. 59:763-769
ISSN:	1869-2044 1590-4261
Popis:	The aim of the present study was to express and evaluate a previously cloned lipase gene. In this study, the cloned gene was subcloned in the pET15bEscherichia coli BL21(DE3) expression system. The expression of the recombinant lipase was induced using 1 mM IPTG for 3 hours. The enzyme activity was measured using p-nitrophenyl-decanoate as substrate. The recombinant lipase showed a molecular weight of 26 kDa by SDS-PAGE. Maximum activity was found at pH 9-10 and 40-50 °C. ZnCl2 at 1, 0.3, 0.1, 0.03, and 0.01 mM concentrations were found to be inhibitory to the enzyme activity and did not improve enzyme thermostability. The recombinant lipase showed an optimum temperature higher than lipase ofBacillus subtilis (its closest relative in primary structure) while similar activity in the alkaline pH.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::63a33e87289b45ce358a5b22d56a8478 https://doi.org/10.1007/bf03179221 Zobrazit plný text záznamu Full text from SpringerLink