15. New Applications of Transketolase: Cascade Reactions for Assay Development

  1. Sergio Riva3 and
  2. Wolf-Dieter Fessner4
  1. Laurence Hecquet1,
  2. Wolf-Dieter Fessner2,
  3. Virgil Hélaine1 and
  4. Franck Charmantray1

Published Online: 27 JUN 2014

DOI: 10.1002/9783527682492.ch15

Cascade Biocatalysis : Integrating Stereoselective and Environmentally Friendly Reactions

Cascade Biocatalysis : Integrating Stereoselective and Environmentally Friendly Reactions

How to Cite

Hecquet, L., Fessner, W.-D., Hélaine, V. and Charmantray, F. (2014) New Applications of Transketolase: Cascade Reactions for Assay Development, in Cascade Biocatalysis : Integrating Stereoselective and Environmentally Friendly Reactions (eds S. Riva and W.-D. Fessner), Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany. doi: 10.1002/9783527682492.ch15

Editor Information

  1. 3

    Istituto di Chimica del, Riconoscimento Molecolare, C. N. R., Via Mario Bianco 9, 20131 Milano, Italy

  2. 4

    Technische Universität, Darmstadt, Department of Organic, Chemistry and Biochemistry, Petersenstr. 22, D-64287 Darmstadt, Germany

Author Information

  1. 1

    Université Blaise Pascal, Institut de Chimie de Clermont-Ferrand (ICCF), UMR CNRS 6296, BP 10448, F-63177, Aubière, France

  2. 2

    Technische Universität Darmstadt, Department of Organic Chemistry and Biochemistry, Petersenstr. 22, D-64287, Darmstadt, Germany

Publication History

  1. Published Online: 27 JUN 2014
  2. Published Print: 23 JUL 2014

ISBN Information

Print ISBN: 9783527335220

Online ISBN: 9783527682492

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Keywords:

  • Transketolase;
  • enzyme;
  • pentose phosphate pathway;
  • thiamine pyrophosphate;
  • ketoses;
  • screening, and selection assays;
  • biocatalysis

Summary

Transketolase (TK; EC 2.2.1.1), a thiamine diphosphate (ThDP)-dependent enzyme, is a key enzyme in the nonoxidative branch of the pentose phosphate pathway. TK catalyzes the stereospecific formation of a C–C bond by a reversible transfer of the C1–C2 ketol unit from a ketose phosphate to an aldose phosphate. TK occurs ubiquitously in all organisms. Microbial TKs have been largely used as biocatalysts for the synthesis of D-threo (3S,4R) ketoses and analogs from nonnatural substrates such as hydroxypyruvate (HPA) as donor and various aldehydes as acceptors. Under these conditions, the reaction becomes irreversible because of the release of carbon dioxide. The modification of the substrate specificity of TKs by rational or random mutagenesis promises to broaden their application range. The screening of large libraries of mutant enzymes requires efficient in vitro high throughput screening (HTS) assays or the direct detection of enzyme activity in host cells using in vivo assays, to obviate the need for cell lysis and purification of enzymes. In addition, some recent studies have shown that human TK is a target in various diseases, such as neurodegenerative diseases, diabetes, and cancer, suggesting new therapeutic approaches. Therefore, the development of rapid, sensitive, and efficient methods for TK activity profiling is also highly desirable to enable the identification of suitable inhibitors. In this context, we have recently developed new assays for TK activity detection. We present the recent in vivo and in vitro TK assays on the basis of cascade reactions involving one or more auxiliary enzymes.