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Biotechnology Letters

, Volume 29, Issue 5, pp 837–841 | Cite as

Production of a recombinant carotenoid cleavage dioxygenase from grape and enzyme assay in water-miscible organic solvents

  • Sandrine Mathieu
  • Frédéric Bigey
  • Jérôme Procureur
  • Nancy Terrier
  • Ziya Günata
Original Research Paper

Abstract

A recombinant carotenoid cleavage dioxygenase from Vitis vinifera L. was produced by Escherichia coli as a fusion with the glutathione-S-transferase (GST) protein under different bacterial growth conditions. The enzyme production was monitored by a GST assay. Addition of Triton X-100 prior to bacterial cell disruption doubled the release of soluble protein. A simple spectrophotometric enzyme assay was developed to measure carotenoid cleavage activity using lutein as substrate. Enzyme activity showed a 26-fold increase with the addition of 10% (v/v) acetone in the reaction mixture.

Keywords

Carotenoid cleavage dioxygenase C13-norisoprenoids Spectrophotometric assay Water-miscible solvents 

Notes

Acknowledgement

We gratefully thank F.X. Sauvage and R. Ratomahenina for their helpful suggestions. This work was supported by a grant from the French “Ministère de l’Education Nationale, de l’Enseignement Supérieur et de la Recherche”.

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Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Sandrine Mathieu
    • 1
  • Frédéric Bigey
    • 1
  • Jérôme Procureur
    • 1
  • Nancy Terrier
    • 2
  • Ziya Günata
    • 1
  1. 1.UMR IR2BENSAM-INRA, Université Montpellier IIMontpellier cedex 1France
  2. 2.UMR SPO, équipe Biologie Intégrative de la Vigne et du RaisinENSAM-INRAMontpellier cedex 1France

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