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A carbohydrate binding module as a diversity-carrying scaffold

Cicortas Gunnarsson, Lavinia LU ; Nordberg Karlsson, Eva LU ; Albrekt, Ann-Sofie LU ; Andersson, M; Holst, Olle LU and Ohlin, Mats LU (2004) In Protein Engineering Design & Selection 17(3). p.213-221
Abstract
The growing field of biotechnology is in constant need of binding proteins with novel properties. Not just binding specificities and affinities but also structural stability and productivity are important characteristics for the purpose of large-scale applications. In order to find such molecules, libraries are created by diversifying naturally occurring binding proteins, which in those cases serve as scaffolds. In this study, we investigated the use of a thermostable carbohydrate binding module, CBM4-2, from a xylanase found in Rhodothermus marinus, as a diversity-carrying scaffold. A combinatorial library was created by introducing restricted variation at 12 positions in the carbohydrate binding site of the CBM4-2. Despite the small size... (More)
The growing field of biotechnology is in constant need of binding proteins with novel properties. Not just binding specificities and affinities but also structural stability and productivity are important characteristics for the purpose of large-scale applications. In order to find such molecules, libraries are created by diversifying naturally occurring binding proteins, which in those cases serve as scaffolds. In this study, we investigated the use of a thermostable carbohydrate binding module, CBM4-2, from a xylanase found in Rhodothermus marinus, as a diversity-carrying scaffold. A combinatorial library was created by introducing restricted variation at 12 positions in the carbohydrate binding site of the CBM4-2. Despite the small size of the library (1.6x10(6) clones), variants specific towards different carbohydrate polymers (birchwood xylan, Avicel and ivory nut mannan) as well as a glycoprotein (human IgG4) were successfully selected for, using the phage display method. Investigated clones showed a high productivity (on average 69 mg of purified protein/l shake flask culture) when produced in Escherichia coli and they were all stable molecules displaying a high melting transition temperature (75.7 +/- 5.3degreesC). All our results demonstrate that the CBM4-2 molecule is a suitable scaffold for creating variants useful in different biotechnological applications. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Protein Engineering Design & Selection
volume
17
issue
3
pages
213 - 221
publisher
Oxford University Press
external identifiers
  • wos:000222797300003
  • scopus:2942748131
ISSN
1741-0126
DOI
10.1093/protein/gzh026
language
English
LU publication?
yes
id
4e49756a-b668-46bb-a83e-985eaa1513df (old id 140613)
date added to LUP
2007-07-02 11:14:02
date last changed
2017-09-03 04:35:35
@article{4e49756a-b668-46bb-a83e-985eaa1513df,
  abstract     = {The growing field of biotechnology is in constant need of binding proteins with novel properties. Not just binding specificities and affinities but also structural stability and productivity are important characteristics for the purpose of large-scale applications. In order to find such molecules, libraries are created by diversifying naturally occurring binding proteins, which in those cases serve as scaffolds. In this study, we investigated the use of a thermostable carbohydrate binding module, CBM4-2, from a xylanase found in Rhodothermus marinus, as a diversity-carrying scaffold. A combinatorial library was created by introducing restricted variation at 12 positions in the carbohydrate binding site of the CBM4-2. Despite the small size of the library (1.6x10(6) clones), variants specific towards different carbohydrate polymers (birchwood xylan, Avicel and ivory nut mannan) as well as a glycoprotein (human IgG4) were successfully selected for, using the phage display method. Investigated clones showed a high productivity (on average 69 mg of purified protein/l shake flask culture) when produced in Escherichia coli and they were all stable molecules displaying a high melting transition temperature (75.7 +/- 5.3degreesC). All our results demonstrate that the CBM4-2 molecule is a suitable scaffold for creating variants useful in different biotechnological applications.},
  author       = {Cicortas Gunnarsson, Lavinia and Nordberg Karlsson, Eva and Albrekt, Ann-Sofie and Andersson, M and Holst, Olle and Ohlin, Mats},
  issn         = {1741-0126},
  language     = {eng},
  number       = {3},
  pages        = {213--221},
  publisher    = {Oxford University Press},
  series       = {Protein Engineering Design & Selection},
  title        = {A carbohydrate binding module as a diversity-carrying scaffold},
  url          = {http://dx.doi.org/10.1093/protein/gzh026},
  volume       = {17},
  year         = {2004},
}