Molecular engineering of ligand-binding proteins is commonly used for identification of variants that display novel specificities. Using this approach to introduce novel specificities into CBMs (carbohydrate-binding modules) has not been extensively explored. Here, we report the engineering of a CBM, CBM4-2 from the Rhodothermus marinus xylanase Xyn10A, and the identification of the X-2 variant. As compared with the wild-type protein, this engineered module displays higher specificity for the polysaccharide xylan, and a lower preference for binding xylo-oligomers rather than binding the natural decorated polysaccharide. The mode of binding of X-2 differs from other xylan-specific CBMs in that it only has one aromatic residue in the binding site that can make hydrophobic interactions with the sugar rings of the ligand. The evolution of CBM4-2 has thus generated a xylan-binding module with different binding properties to those displayed by CBMs available in Nature.
Skip Nav Destination
Article navigation
September 2007
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
- PDF Icon PDF LinkEditorial Board
Research Article|
August 13 2007
Novel xylan-binding properties of an engineered family 4 carbohydrate-binding module
Lavinia Cicortas Gunnarsson;
Lavinia Cicortas Gunnarsson
*Department of Immunotechnology, Lund University, BMC D13, SE-221 84 Lund, Sweden
Search for other works by this author on:
Cedric Montanier;
Cedric Montanier
†Institute for Cell and Molecular Biosciences, University of Newcastle upon Tyne, Framlington Place, Newcastle upon Tyne NE2 4HH, U.K.
Search for other works by this author on:
Richard B. Tunnicliffe;
Richard B. Tunnicliffe
‡Department of Molecular Biology and Biotechnology, University of Sheffield, Firth Court, Western Bank, Sheffield S10 2TN, U.K.
Search for other works by this author on:
Mike P. Williamson;
Mike P. Williamson
‡Department of Molecular Biology and Biotechnology, University of Sheffield, Firth Court, Western Bank, Sheffield S10 2TN, U.K.
Search for other works by this author on:
Harry J. Gilbert;
Harry J. Gilbert
†Institute for Cell and Molecular Biosciences, University of Newcastle upon Tyne, Framlington Place, Newcastle upon Tyne NE2 4HH, U.K.
Search for other works by this author on:
Eva Nordberg Karlsson;
Eva Nordberg Karlsson
§Department of Biotechnology, Lund University, P.O. Box 124, SE-221 00 Lund, Sweden
Search for other works by this author on:
Mats Ohlin
Mats Ohlin
1
*Department of Immunotechnology, Lund University, BMC D13, SE-221 84 Lund, Sweden
1To whom correspondence should be addressed (email mats.ohlin@immun.lth.se).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
January 24 2007
Revision Received:
May 09 2007
Accepted:
May 16 2007
Accepted Manuscript online:
May 16 2007
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2007 Biochemical Society
2007
Biochem J (2007) 406 (2): 209–214.
Article history
Received:
January 24 2007
Revision Received:
May 09 2007
Accepted:
May 16 2007
Accepted Manuscript online:
May 16 2007
Citation
Lavinia Cicortas Gunnarsson, Cedric Montanier, Richard B. Tunnicliffe, Mike P. Williamson, Harry J. Gilbert, Eva Nordberg Karlsson, Mats Ohlin; Novel xylan-binding properties of an engineered family 4 carbohydrate-binding module. Biochem J 1 September 2007; 406 (2): 209–214. doi: https://doi.org/10.1042/BJ20070128
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.