Surface display of Zymomonas mobilis levansucrase by using the ice-nucleation protein of Pseudomonas syringae

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Title
Surface display of Zymomonas mobilis levansucrase by using the ice-nucleation protein of Pseudomonas syringae
Author(s)
Heung Chae Jung; Jean M Lebeault; Jae Gu Pan
Bibliographic Citation
Nature Biotechnology, vol. 16, no. 6, pp. 576-580
Publication Year
1998
Abstract
The ice-nucleation protein (Inp) is a glycosyl phosphatidylinositol- anchored outer membrane protein found in some Gram-negative bacteria. Using Pseudornonas syringae Inp as an anchoring motif, we investigated the functional display of a foreign protein, Zymomonas mobilis levansucrase (LevU), on the surface of Escherichia coli. The cells expressing Inp-LevU were found to retain both the ice-nucleation and whole-cell levansucrase enzyme activities, indicating the functional expression of Inp-LevU hybrid protein on the cell surface. The surface localization was further verified by immunofluorescence microscopy, fluorescence-activated cell sorting flow cytometry and immunogold electron microscopical examination. No growth inhibition or changes in the outer membrane integrity were observed upon the induction of fusion protein synthesis. Viability of the cells was also maintained over 48 hours in the stationary phase. Surface-displayed levansucrases were found to be resistant to the externally added proteases unless the cells were treated with EDTA. When the levansucrase-displayed cells were used as the enzyme source, levan (44 g/L) was efficiently synthesized from sucrose (130 g/L) with 34% (wt/wt) conversion yield, generating glucose (65 g/L) as a by-product.
Keyword
BioconversionGPI anchorProtein display
ISSN
0733-222X
Publisher
Springer-Nature Pub Group
DOI
http://dx.doi.org/10.1038/nbt0698-576
Type
Article
Appears in Collections:
Division of Bio Technology Innovation > SME Support Center > 1. Journal Articles
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