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Plant and Food Carotenoids pp 63–74Cite as

Determination of In Vitro and In Vivo Activities of Plant Carotenoid Cleavage Oxygenases

Part of the Methods in Molecular Biology book series (MIMB,volume 2083)

Abstract

Carotenoid cleavage products, apocarotenoids, are biologically active compounds exerting important functions as chromophore, hormones, signaling molecules, volatiles, and pigments. Apocarotenoids are generally synthesized by the carotenoid cleavage dioxygenases (CCDs) that comprise a ubiquitous family of enzymes. The activity of plant CCDs was unraveled more than 20 years ago, with the characterization of the maize VP14, the first identified CCD. The protocol developed to determine the activity of this enzyme in vitro is still being used, with minor modifications. In addition, in vivo procedures have been developed during these years, mainly based on the exploitation of Escherichia coli cells engineered to produce specific carotenoid substrates. Further, technological developments have led to significant improvements, contributing to a more efficient detection of the reaction products. This chapter provides an updated set of detailed protocols suitable for the in vitro and in vivo characterization of the activities of CCDs, starting from well-established methods.

Key words

  • Apocarotenoids
  • Carotenoids
  • CCD
  • Enzymatic activity

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Authors and Affiliations

  1. Departamento de Ciencia y Tecnología Agroforestal y Genética, Instituto Botánico, Universidad de Castilla-La Mancha, Albacete, Spain

    Lourdes Gómez-Gómez & Oussama Ahrazem

  2. Italian National Agency for New Technologies, Energy, and Sustainable Development, Casaccia Research Centre, Rome, Italy

    Gianfranco Diretto

  3. The Bioactives Lab, Division of Biological and Environmental Science and Engineering, King Abdullah University of Science and Technology, Thuwal, Saudi Arabia

    Salim Al-Babili

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  1. Lourdes Gómez-Gómez
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  2. Gianfranco Diretto
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  3. Oussama Ahrazem
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  4. Salim Al-Babili
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Correspondence to Lourdes Gómez-Gómez .

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Editors and Affiliations

  1. CSIC-IRTA-UAB-UB, Centre for Research in Agricultural Genomics (CRAG), Barcelona, Spain

    Manuel Rodríguez-Concepción

  2. Institute for Biology II, University of Freiburg, Freiburg, Germany

    Ralf Welsch

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Gómez-Gómez, L., Diretto, G., Ahrazem, O., Al-Babili, S. (2020). Determination of In Vitro and In Vivo Activities of Plant Carotenoid Cleavage Oxygenases. In: Rodríguez-Concepción, M., Welsch, R. (eds) Plant and Food Carotenoids. Methods in Molecular Biology, vol 2083. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9952-1_5

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