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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 14, No. 3, 2011
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Bioline Code: ej11029
Full paper language: English
Document type: Review Article
Document available free of charge
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Electronic Journal of Biotechnology, Vol. 14, No. 3, 2011
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Recombinant expression and refolding of the c-type lysozyme from Spodoptera litura in E. coli
Kim, Jong-Wan & Yoe, Sung Moon
Abstract
The chicken-type lysozyme of the insect Spodoptera litura (SLLyz) is a polypeptide of 121 amino acids containing four disulfide bridges and 17 rare codons and participates in innate defense as an anti-bacterial enzyme. The recombinant S. litura lysozyme (rSLLyz) expressed as a C-terminal fusion protein with glutathione S-transferase (GST) in Rosetta(DE3) Singles. The protein was produced as an inclusion body which was solubilized in 8 M urea, renatured by on-column refolding, and purified by reversed-phase chromatography to 95% purity. The purified rSLLyz demonstrated antibacterial activity against B. megateriumI confirmed by inhibition zone assay. The overexpression and refolding strategy described in this study will provide a reliable technique for maximizing production and purification of proteins expressed as inclusion bodies in E. coli.
Keywords
antibacterial activity, inclusion body, lysozyme, on-column refolding, recombinant expression, Spodoptera litura
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© Copyright 2011 by Universidad Católica de Valparaíso -- Chile Alternative site location: http://www.ejbiotechnology.info
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