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Cloning of the astaxanthin synthase gene from Xanthophyllomyces dendrorhous (Phaffia rhodozyma) and its assignment as a β-carotene 3-hydroxylase/4-ketolase

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Abstract

A gene has been cloned from Xanthophyllomyces dendrorhous by complementation of astaxanthin formation in a β-carotene accumulating mutant. It consists of 3,166 bp and contains 17 introns. For the β-carotene mutant ATCC 96815, a single point mutation in the splicing sequence of intron 8 was found. The resulting improper splicing of the mRNA results in an inactive protein. The cDNA of this β-carotene oxygenase encodes a cytochrome P450 monooxygenase belonging to the 3A subfamily. P450-specific domains were identified including a cytochrome P450 and an oxygen binding motif. Electrons are provided by a cytochrome P450 reductase. Functional characterization of the enzyme by genetic modification of X. dendrorhous demonstrated that this P450 monooxygenase is multifunctional catalyzing all steps from β-carotene to astaxanthin formation by oxygenation of carbon 3 and 4. The reaction sequence is first 4-ketolation of β-carotene followed by 3-hydroxylation. A hydroxylation mechanism at allylic carbon atoms has been proposed for the generation of 4-keto and 3-hydroxy groups at both β-ionone ends.

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Acknowledgements

This work was supported by grant QLK1-CT-2001-00780 from the European Commission.

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Author notes

  1. Tatsuo Hoshino

    Present address: Tamagawa University Research Institute, 6-1-1 Tamagawa-Gakuen, Machida, Tokyo, Japan

  2. Hans Visser

    Present address: Dyadic Nederland BV, Utrechtseweg 48, 3704, Zeist, HE, The Netherlands

Authors and Affiliations

  1. Department of Applied Microbiology, Nippon Roche Research Center, 247-8530, Kamakura, Kanagawa, Japan

    Kazuyuki Ojima, Yutaka Setoguchi, Kazuyuki Tabata & Tatsuo Hoshino

  2. J. W. Goethe Universität, Molecular Biosciences 213, P.O. Box 111932, 60054, Frankfurt, Germany

    Jürgen Breitenbach & Gerhard Sandmann

  3. Section of Fungal Genomics, Wageningen University, Dreijenlaan 2, 6703, Wageningen, HA, The Netherlands

    Hans Visser & Johan van den Berg

Authors
  1. Kazuyuki Ojima
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  2. Jürgen Breitenbach
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Correspondence to Gerhard Sandmann.

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Communicated by P. Punt

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Ojima, K., Breitenbach, J., Visser, H. et al. Cloning of the astaxanthin synthase gene from Xanthophyllomyces dendrorhous (Phaffia rhodozyma) and its assignment as a β-carotene 3-hydroxylase/4-ketolase. Mol Genet Genomics 275, 148–158 (2006). https://doi.org/10.1007/s00438-005-0072-x

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  • DOI: https://doi.org/10.1007/s00438-005-0072-x

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