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標題: A system for purification of recombinant proteins in Escherichia coli via artificial oil bodies constituted with their oleosin-fused polypeptides
作者: Peng, C.C.
Chen, J.C.F.
Shyu, D.J.H.
Chen, M.J.
Tzen, J.I.C.
關鍵字: artificial oil body;green fluorescent protein;oleosin;factor Xa;sesame;fusions;seeds;biogenesis;caleosin;surface;cloning;tag
Project: Journal of Biotechnology
期刊/報告no:: Journal of Biotechnology, Volume 111, Issue 1, Page(s) 51-57.
An expression/purification system was developed using artificial oil bodies (AOB) as carriers for producing recombinant proteins. A target protein, green fluorescent protein (GFP), was firstly expressed in Escherichia coli as an insoluble recombinant protein fused to oleosin, a unique structural protein of seed oil bodies, by a linker sequence susceptible to factor Xa cleavage. Artificial oil bodies were constituted with triacylglycerol, phospholipid, and the insoluble recombinant protein, oleosin-Xa-GFP. After centrifugation, the oleosin-fused GFP was exclusively found on the surface of artificial oil bodies presumably with correct folding to emit fluorescence under excitation. Proteolytic cleavage with factor Xa separated soluble GFP from oleosin embedded in the artificial oil bodies; thus after re-centrifugation, GFP of high yield and purity was harvested simply by concentrating the ultimate supernatant. (C) 2004 Elsevier B.V. All rights reserved.
ISSN: 0168-1656
DOI: 10.1016/j.jbiotec.2004.03.013
Appears in Collections:生物科技學研究所

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