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

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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.

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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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Authors and Affiliations

  1. UMR IR2B, ENSAM-INRA, Université Montpellier II, 34060, Montpellier cedex 1, France

    Sandrine Mathieu, Frédéric Bigey, Jérôme Procureur & Ziya Günata

  2. UMR SPO, équipe Biologie Intégrative de la Vigne et du Raisin, ENSAM-INRA, 34060, Montpellier cedex 1, France

    Nancy Terrier

Authors
  1. Sandrine Mathieu
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  2. Frédéric Bigey
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  3. Jérôme Procureur
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  4. Nancy Terrier
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  5. Ziya Günata
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Correspondence to Ziya Günata.

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Mathieu, S., Bigey, F., Procureur, J. et al. Production of a recombinant carotenoid cleavage dioxygenase from grape and enzyme assay in water-miscible organic solvents. Biotechnol Lett 29, 837–841 (2007). https://doi.org/10.1007/s10529-007-9315-8

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  • DOI: https://doi.org/10.1007/s10529-007-9315-8

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