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|Title:||Functional display of family 11 endoxylanases on the surface of phage M13||Authors:||Beliën, T.
Van den Brande, K
Van Campenhout, S
|Issue Date:||2005||Publisher:||ELSEVIER SCIENCE BV||Source:||JOURNAL OF BIOTECHNOLOGY, 115(3). p. 249-260||Abstract:||Two family 11 endoxylanases (EC 184.108.40.206) 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.||Notes:||Katholieke Univ Leuven, Lab Gene Technol, B-3001 Heverlee, Belgium. Limburgs Univ Ctr, Inst Biomed Res, B-3590 Diepenbeek, Belgium.Van Campenhout, S, Katholieke Univ Leuven, Lab Gene Technol, Kasteelpk Arenberg 21, B-3001 Heverlee, Belgium.firstname.lastname@example.org||Keywords:||Aspergillus niger endoxylanase; Bacillus subtilis endoxylanase; endoxylanase inhibitors; phage display||Document URI:||http://hdl.handle.net/1942/2139||ISSN:||0168-1656||e-ISSN:||1873-4863||DOI:||10.1016/j.jbiotec.2004.08.013||ISI #:||000226457400003||Category:||A1||Type:||Journal Contribution||Validations:||ecoom 2006|
|Appears in Collections:||Research publications|
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