Rational and combinatorial engineering of the glucan synthesizing enzyme amylosucrase

C. Albenne; B.A. Van Der Veen; G. Potocki-Véronèse; G. Joucla; L. Skov; Osman Asghar Mirza; M. Gajhede; P. Monsan; M. Remaud-Simeon

doi:10.1080/10242420310001618537

Rational and combinatorial engineering of the glucan synthesizing enzyme amylosucrase

C. Albenne, B.A. Van Der Veen, G. Potocki-Véronèse, G. Joucla, L. Skov, Osman Asghar Mirza, M. Gajhede, P. Monsan, M. Remaud-Simeon

3 Citations (Scopus)

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.

Original language	English
Journal	Biocatalysis and Biotransformation
Volume	21
Issue number	4-5
Pages (from-to)	271-277
Number of pages	7
ISSN	1024-2422
DOIs	https://doi.org/10.1080/10242420310001618537
Publication status	Published - 1 Aug 2003

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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.",

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AU - Albenne, C.

AU - Van Der Veen, B.A.

AU - Potocki-Véronèse, G.

AU - Joucla, G.

AU - Skov, L.

AU - Mirza, Osman Asghar

AU - Gajhede, M.

AU - Monsan, P.

AU - Remaud-Simeon, M.

PY - 2003/8/1

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N2 - 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.

AB - 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.

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Rational and combinatorial engineering of the glucan synthesizing enzyme amylosucrase

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