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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/140613
- author
- Cicortas Gunnarsson, Lavinia
LU
; Nordberg Karlsson, Eva
LU
; Albrekt, Ann-Sofie
LU
; Andersson, M
; Holst, Olle
LU
and Ohlin, Mats
LU
- organization
- publishing date
- 2004
- 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
- project
- Designed carbohydrate binding modules and molecular probes
- language
- English
- LU publication?
- yes
- id
- 4e49756a-b668-46bb-a83e-985eaa1513df (old id 140613)
- date added to LUP
- 2016-04-01 16:19:39
- date last changed
- 2022-01-28 18:55:52
@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}},
doi = {{10.1093/protein/gzh026}},
volume = {{17}},
year = {{2004}},
}