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Title 

Functional display of active tetrameric ß-galactosidase using Bacillus subtilis spore display system

Authors 

B Y HwangJae Gu PanB G KimJ H Kim

Publisher 

American Scientific Publishers

Issue Date 

2013

Citation 

Journal of Nanoscience and Nanotechnology, vol. 13, no. 3, pp. 2313-2319

Keywords 

ß-GalactosidaseBacillus subtilisSporeSurface display

Abstract 

For the functional bacterial surface display of active enzyme of multimeric form, which is generally impossible due to molecular assembly of the monomer subunit subsequent to the secretion of displayed target protein outside the cell, a new surface display system based on B. subtilis spore was developed. Using cotE and cotG of B. subtilis as anchoring motives, ß-galactosidase, which is active in tetrameric form, was functionally displayed on the surface of B. subtilis spore. The surface localization of ß-galactosidase was verified by Miller assay of purified spore, protease accessibility test of purified spore, and flow cytometric analysis of spore expressing ß-galactosidase. While B. subtilis spore wall integrity, examined by lysozyme and heat treatments, was affected by the incorporation of CotE-LacZ fusion protein, it was not affected by the incorporation of CotG-lacZ fusion. Heat stability of displayed protein was similar with that of free enzyme.

ISSN 

1533-4880

Link 

http://dx.doi.org/10.1166/jnn.2013.6889

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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