Subscribe to thousands of academic journals for just $40/month
Read and share the articles you need for your research, all in one place.

Rational and Combinatorial Engineering of the Glucan Synthesizing Enzyme Amylosucrase

Biocatalysis and Biotransformation , Volume 21 (4-5) – Jan 1, 2003

Details

Publisher
Informa UK Ltd
Copyright
© 2003 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted
Subject
ORIGINAL ARTICLE
ISSN
1024-2422
eISSN
1029-2446
D.O.I.
10.1080/10242420310001618537
Publisher site
See Article on Publisher Site

Preview Only

Expand Tray Hide Tray

Rational and Combinatorial Engineering of the Glucan Synthesizing Enzyme Amylosucrase

Abstract

Rational engineering of amylosucrase required detailed investigations of the molecular basis of catalysis. Biochemical characterization of the enzyme coupled to structural analyses enabled the polymerization mechanism to be elucidated. This provided key information for successfully changing amylosucrase to a hydrolase or an improved polymerase using site-directed mutagenesis. In parallel, a combinatorial approach was developed to further improve the catalyst. The method, based on random mutagenesis and recombination of parent sequences, has generated libraries of variants. These were searched for improved polymer formation using a first step of selection followed by a screening test including a double detection method. Several mutants with desired properties have been isolated, their sequences revealed that they could not have been designed following a rational approach.
Loading next page...

Preview Only. This article cannot be rented because we do not currently have permission from the publisher.

 
/lp/informa-healthcare/rational-and-combinatorial-engineering-of-the-glucan-synthesizing-dWwZyTuMLV