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Molecular Genetics and Genomics

, Volume 275, Issue 2, pp 148–158 | Cite as

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

  • Kazuyuki Ojima
  • Jürgen Breitenbach
  • Hans Visser
  • Yutaka Setoguchi
  • Kazuyuki Tabata
  • Tatsuo Hoshino
  • Johan van den Berg
  • Gerhard SandmannEmail author
Original Paper

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.

Keywords

Astaxanthin synthase Cytochrome P450 Genetic complementation Monooxygenase 

Notes

Acknowledgements

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

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Kazuyuki Ojima
    • 1
  • Jürgen Breitenbach
    • 2
  • Hans Visser
    • 3
    • 5
  • Yutaka Setoguchi
    • 1
  • Kazuyuki Tabata
    • 1
  • Tatsuo Hoshino
    • 1
    • 4
  • Johan van den Berg
    • 3
  • Gerhard Sandmann
    • 2
    Email author
  1. 1.Department of Applied MicrobiologyNippon Roche Research CenterKamakura, KanagawaJapan
  2. 2.J. W. Goethe UniversitätFrankfurtGermany
  3. 3.Section of Fungal GenomicsWageningen UniversityWageningenThe Netherlands
  4. 4.Tamagawa University Research InstituteMachida, TokyoJapan
  5. 5.Dyadic Nederland BVZeistThe Netherlands

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