Novel xylan-binding properties of an engineered family 4 carbohydrate-binding module

Cicortas Gunnarsson, LaviniaLund University,Lunds universitet,Institutionen för immunteknologi,Institutioner vid LTH,Lunds Tekniska Högskola,Department of Immunotechnology,Departments at LTH,Faculty of Engineering, LTH,Department of Immunotechnology, Lund University, Lund, Sweden (författare)

Funding Agency:Biotechnology and Biological Sciences Research Council BB/C005074/1
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, CBM42 from the Rhodothermits marinus xylanase Xyn10A, and the identification of the X-2 variant. As compared with the wildtype 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.

NATURVETENSKAP Biologi Biokemi och molekylärbiologi hsv//swe
NATURAL SCIENCES Biological Sciences Biochemistry and Molecular Biology hsv//eng
thermodynamics
molecular engineering
carbohydrate-binding module
aromatic residue
binding specificity
xylan
aromatic residue; binding specificity; carbohydrate-binding module; molecular engineering; thermodynamics; xylan

Montanier, CedricInstitute for Cell and Molecular Biosciences, University of Newcastle upon Tyne, Newcastle upon Tyne, UK (författare)
Tunnicliffe, Richard B.Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield, UK (författare)
Williamson, Mike R. (författare)
Gilbert, Harry J.Institute for Cell and Molecular Biosciences, University of Newcastle upon Tyne, Newcastle upon Tyne, UK (författare)
Nordberg Karlsson, EvaLund University,Lunds universitet,Bioteknik,Centrum för tillämpade biovetenskaper,Kemiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Biotechnology,Center for Applied Life Sciences,Department of Chemistry,Departments at LTH,Faculty of Engineering, LTH,Department of Biotechnology, Lund University, Lund, Sweden(Swepub:lu)biot-eno (författare)
Ohlin, MatsLund University,Lunds universitet,Institutionen för immunteknologi,Institutioner vid LTH,Lunds Tekniska Högskola,Department of Immunotechnology,Departments at LTH,Faculty of Engineering, LTH,Department of Immunotechnology, Lund University, Lund, Sweden(Swepub:lu)immt-moh (författare)
Williamson, Mike PDepartment of Molecular Biology and Biotechnology, University of Sheffield, Sheffield, UK (författare)
Institutionen för immunteknologiInstitutioner vid LTH (creator_code:org_t)

Av författaren/redakt...: Cicortas Gunnars ...; Montanier, Cedri ...; Tunnicliffe, Ric ...; Williamson, Mike ...; Gilbert, Harry J ...; Nordberg Karlsso ...; visa fler...; Ohlin, Mats; Williamson, Mike ...; visa färre...

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