A fluorogenic assay for transketolase from Saccharomyces cerevisiae

In order to generate transketolase (TK) with new or improved properties by in vitro evolution, an efficient screening system is an absolute prerequisite for identifying the evolved enzyme variants. We report here an assay allowing us to detect wild type TK activity in vitro by fluorescence. We exami...

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Published inTetrahedron letters Vol. 44; no. 4; pp. 827 - 830
Main Authors Sevestre, Aurélie, Hélaine, Virgil, Guyot, Ghislain, Martin, Christine, Hecquet, Laurence
Format Journal Article
LanguageEnglish
Published OXFORD Elsevier Ltd 2003
Elsevier
Subjects
Chemistry
Chemistry, Organic
Life Sciences
Physical Sciences
Science & Technology
ENANTIOSELECTIVITY
ALDOLASE
DIRECTED EVOLUTION
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Abstract In order to generate transketolase (TK) with new or improved properties by in vitro evolution, an efficient screening system is an absolute prerequisite for identifying the evolved enzyme variants. We report here an assay allowing us to detect wild type TK activity in vitro by fluorescence. We examined the use of the fluorogenic compound 1 as donor substrate of TK, which is itself non fluorescent but releases a fluorescent product: umbelliferone. Graphic
AbstractList In order to generate transketolase (TK) with new or improved properties by in vitro evolution, an efficient screening system is an absolute prerequisite for identifying the evolved enzyme variants. We report here an assay allowing us to detect wild type TK activity in vitro by fluorescence. We examined the use of the fluorogenic compound I as donor substrate of TK, which is itself non fluorescent but releases a fluorescent product: umbelliferone. (C) 2003 Elsevier Science Ltd. All rights reserved.
In order to generate transketolase (TK) with new or improved properties by in vitro evolution, an efficient screening system is an absolute prerequisite for identifying the evolved enzyme variants. We report here an assay allowing us to detect wild type TK activity in vitro by fluorescence. We examined the use of the fluorogenic compound I as donor substrate of TK, which is itself non fluorescent but releases a fluorescent product: umbelliferone.
In order to generate transketolase (TK) with new or improved properties by in vitro evolution, an efficient screening system is an absolute prerequisite for identifying the evolved enzyme variants. We report here an assay allowing us to detect wild type TK activity in vitro by fluorescence. We examined the use of the fluorogenic compound 1 as donor substrate of TK, which is itself non fluorescent but releases a fluorescent product: umbelliferone. Graphic
ArticleNumber 0040
Author Sevestre, Aurélie
Guyot, Ghislain
Hélaine, Virgil
Martin, Christine
Hecquet, Laurence
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  givenname: Ghislain
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  givenname: Christine
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  organization: Université Blaise Pascal, Laboratoire de Synthèse et Etude de Systèmes à Intérêt Biologique, UMR 6504, 63177 Aubière Cedex, France
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Issue 4
Keywords ENANTIOSELECTIVITY
ALDOLASE
DIRECTED EVOLUTION
Language English
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Snippet In order to generate transketolase (TK) with new or improved properties by in vitro evolution, an efficient screening system is an absolute prerequisite for...
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SubjectTerms Chemistry
Chemistry, Organic
Life Sciences
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Title A fluorogenic assay for transketolase from Saccharomyces cerevisiae
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