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Increasing the thermostability of sucrose phosphorylase by a combination of sequence- and structure-based mutagenesis

An Cerdobbel UGent, Karel De Winter UGent, Dirk Aerts UGent, Remko Kuipers, Henk-Jan Joosten, Wim Soetaert UGent and Tom Desmet UGent (2011) PROTEIN ENGINEERING DESIGN & SELECTION. 24(11). p.829-834
abstract
Sucrose phosphorylase is a promising biocatalyst for the glycosylation of a wide variety of acceptor molecules, but its low thermostability is a serious drawback for industrial applications. In this work, the stability of the enzyme from Bifidobacterium adolescentis has been significantly improved by a combination of smart and rational mutagenesis. The former consists of substituting the most flexible residues with amino acids that occur more frequently at the corresponding positions in related sequences, while the latter is based on a careful inspection of the enzyme's crystal structure to promote electrostatic interactions. In this way, a variant enzyme could be created that contains six mutations and whose half-life at the industrially relevant temperature of 60 degrees C has more than doubled compared with the wild-type enzyme. An increased stability in the presence of organic co-solvents could also be observed, although these effects were most noticeable at low temperatures.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
DIRECTED EVOLUTION, ENHANCED PROTEIN THERMOSTABILITY, thermostability, sucrose phosphorylase, enzyme engineering, solvent stability, RATIONAL DESIGN, STABILITY, ENZYME, MUTATIONS
journal title
PROTEIN ENGINEERING DESIGN & SELECTION
Protein Eng. Des. Sel.
volume
24
issue
11
pages
829 - 834
Web of Science type
Article
Web of Science id
000296132800003
JCR category
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
JCR impact factor
2.937 (2011)
JCR rank
50/157 (2011)
JCR quartile
2 (2011)
ISSN
1741-0126
DOI
10.1093/protein/gzr042
project
Biotechnology for a sustainable economy (Bio-Economy)
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
2121933
handle
http://hdl.handle.net/1854/LU-2121933
date created
2012-05-30 13:59:05
date last changed
2013-02-27 13:06:46
@article{2121933,
  abstract     = {Sucrose phosphorylase is a promising biocatalyst for the glycosylation of a wide variety of acceptor molecules, but its low thermostability is a serious drawback for industrial applications. In this work, the stability of the enzyme from Bifidobacterium adolescentis has been significantly improved by a combination of smart and rational mutagenesis. The former consists of substituting the most flexible residues with amino acids that occur more frequently at the corresponding positions in related sequences, while the latter is based on a careful inspection of the enzyme's crystal structure to promote electrostatic interactions. In this way, a variant enzyme could be created that contains six mutations and whose half-life at the industrially relevant temperature of 60 degrees C has more than doubled compared with the wild-type enzyme. An increased stability in the presence of organic co-solvents could also be observed, although these effects were most noticeable at low temperatures.},
  author       = {Cerdobbel, An and De Winter, Karel and Aerts, Dirk and Kuipers, Remko and Joosten, Henk-Jan and Soetaert, Wim and Desmet, Tom},
  issn         = {1741-0126},
  journal      = {PROTEIN ENGINEERING DESIGN \& SELECTION},
  keyword      = {DIRECTED EVOLUTION,ENHANCED PROTEIN THERMOSTABILITY,thermostability,sucrose phosphorylase,enzyme engineering,solvent stability,RATIONAL DESIGN,STABILITY,ENZYME,MUTATIONS},
  language     = {eng},
  number       = {11},
  pages        = {829--834},
  title        = {Increasing the thermostability of sucrose phosphorylase by a combination of sequence- and structure-based mutagenesis},
  url          = {http://dx.doi.org/10.1093/protein/gzr042},
  volume       = {24},
  year         = {2011},
}

Chicago
Cerdobbel, An, Karel De Winter, Dirk Aerts, Remko Kuipers, Henk-Jan Joosten, Wim Soetaert, and Tom Desmet. 2011. “Increasing the Thermostability of Sucrose Phosphorylase by a Combination of Sequence- and Structure-based Mutagenesis.” Protein Engineering Design & Selection 24 (11): 829–834.
APA
Cerdobbel, A., De Winter, K., Aerts, D., Kuipers, R., Joosten, H.-J., Soetaert, W., & Desmet, T. (2011). Increasing the thermostability of sucrose phosphorylase by a combination of sequence- and structure-based mutagenesis. PROTEIN ENGINEERING DESIGN & SELECTION, 24(11), 829–834.
Vancouver
1.
Cerdobbel A, De Winter K, Aerts D, Kuipers R, Joosten H-J, Soetaert W, et al. Increasing the thermostability of sucrose phosphorylase by a combination of sequence- and structure-based mutagenesis. PROTEIN ENGINEERING DESIGN & SELECTION. 2011;24(11):829–34.
MLA
Cerdobbel, An, Karel De Winter, Dirk Aerts, et al. “Increasing the Thermostability of Sucrose Phosphorylase by a Combination of Sequence- and Structure-based Mutagenesis.” PROTEIN ENGINEERING DESIGN & SELECTION 24.11 (2011): 829–834. Print.