ISSN: 0973-7510

E-ISSN: 2581-690X

Haoran Yu1#, Yanjie Liu1#, Liujie Huo1, Xinya Liang2, Guangbo Kang3 and He Huang1
1School of Chemical Engineering and Technology, Tianjin University, Tianjin – 300072, China.
2Ningbo Green Biotechnology Development Corporation, Ningbo – 305040, China.
3College of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin – 300457, China.
J Pure Appl Microbiol. 2013;7(Spl. Edn.: November):679-685
© The Author(s). 2013
Received: 27/09/2013 | Accepted: 04/11/2013 | Published: 30/11/2013
Abstract

Luciferase catalyzes luciferin to emit yellow green fluorescence in the presence of ATP and Mg2+. Based on such a feature, it played an important role in many scientific and industrial fields. In the study, a simple and rapid expression and purification method of recombinant luciferase was developed. The high expression vector pET28a-luc was constructed and successfully induced to express soluble luciferase. The luciferase was separated and purified through immobilized metal ion affinity chromatography (IMAC). After optimizing expression conditions, 45 mg recombinant luciferase with high specific activity, 4.04±0.50×1012 RLU/mg was achieved per liter of cell culture. It turned out that lower shaking speed and longer induction time were more effective for high yield of soluble protein. Circular dichroism (CD) and fluorescence spectroscopy were performed to verify the proper fold of the recombinant target protein.

Keywords

Luciferase, Prokaryotic expression, Chromatography purification

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