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Title: Functional display of family 11 endoxylanases on the surface of phage M13
Authors: Belien, Tim ×
Hertveldt, Kirsten
Van den Brande, K
Robben, Johan
Van Campenhout, Steven
Volckaert, Guido #
Issue Date: 2005
Publisher: Elsevier science bv
Series Title: Journal of biotechnology vol:115 issue:3 pages:249-260
Abstract: Two family 11 endoxylanases (EC were functionally displayed on the surface of bacteriophage M13. The genes encoding endo-1,4-xylanase I from Aspergillus niger (Ex1A) and endo-1,4-xylanase A from Bacillus subtilis (XynA) were fused to the gene encoding the minor coat protein g3p in phagemid vector pHOS31. Phage rescue resulted in functional monovalent display of the enzymes as was demonstrated by three independent tests. Firstly, purified recombinant phage particles showed a clear hydrolytic activity in an activity assay based on insoluble, chromagenic arabinoxylan substrate. Secondly, specific binding of endoxylanase displaying phages to immobilized endoxylanase inhibitors was demonstrated by interaction ELISA. Finally, two rounds of selection and amplification in a biopanning procedure against immobilized endoxylanase inhibitor were performed. Phages displaying endoxylanases were strongly enriched from background phages displaying unrelated proteins. These results open perspectives to use phage display for analysing protein-protein interactions at the interface between endoxylanases and their inhibitors. In addition, this technology should enable engineering of endoxylanases into novel variants with altered binding properties towards endoxylanase inhibitors. (C) 2004 Elsevier B.V. All rights reserved.
ISSN: 0168-1656
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Division of Gene Technology (-)
Centre for Food and Microbial Technology
Biochemistry, Molecular and Structural Biology Section
× corresponding author
# (joint) last author

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