Molecular engineering of a thermostable carbohydrate-binding module

Summary, in English

Structure-function studies are frequently practiced on the very diverse group of natural carbohydrate-binding modules in order to understand the target recognition of these proteins. We have taken a step further in the study of carbohydrate-binding modules and created variants with novel binding properties by molecular engineering of one such molecule of known 3D-structure. A combinatorial library was created from the sequence encoding a thermostable carbohydrate-binding module, CBM4-2 from a Rhodothermus marinus xylanase, and phage-display technology was successfully used for selection of variants with specificity towards different carbohydrate polymers (birchwood xylan, Avicel (TM), ivory nut mannan and recently also xyloglucan), as well as towards a glycoprotein (human IgG4). Our work not only generated a number of binders with properties that would suite a range of biotechnological applications, but analysis of selected binders also helped us to identify residues important for their specificities.

• Författare: Lavinia Cicortas Gunnarsson; Eva Nordberg Karlsson; Mats Andersson; Olle Holst; Mats Ohlin
• Avdelning/ar: Institutionen för immunteknologi; Bioteknik
• Publiceringsår: 2006
• Språk: Engelska
• Sidor: 31-37
• Publikation/Tidskrift/Serie: Biocatalysis and Biotransformation
• Volym: 24
• Issue: 1-2
• Länkar: http://dx.doi.org/10.1080/10242420500518516
• Dokumenttyp: Konferensbidrag
• Förlag: Taylor & Francis
• Ämne: Immunology in the medical area; Industrial Biotechnology
• Nyckelord: carbohydrate-binding module; binding specificity; combinatorial; library; molecular engineering; phage-display; protein scaffold
• Conference name: Sixth Carbohydrate Bioengineering Meeting
• Conference date: 2005-04-03 - 2005-04-06
• Conference place: Barcelona, Spain
• Status: Published
• Projekt: Designed carbohydrate binding modules and molecular probes
• ISBN/ISSN/Övrigt: ISSN: 1024-2422; ISSN: 1029-2446