Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 428589
Title Increasing the thermostability of sucrose phosphorylase by a combination of sequence- and structure-based mutagenesis
Author(s) Cerdobbel, A.; Winter, K. De; Aerts, D.; Kuipers, R.K.P.; Joosten, H.J.; Soetaert, W.; Desmet, T.
Source Protein Engineering, Design & Selection 24 (2011)11. - ISSN 1741-0126 - p. 829 - 834.
DOI https://doi.org/10.1093/protein/gzr042
Department(s) Microbiological Laboratory
VLAG
Publication type Refereed Article in a scientific journal
Publication year 2011
Keyword(s) enhanced protein thermostability - directed evolution - rational design - stability - enzyme - mutations
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.
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