Cloning of the Astaxanthin Synthase Gene from Xanthophyllomyces Dendrorhous (Phaffia Rhodozyma) and Its Assignment As a Beta-carotene 3-hydroxylase/4-ketolase

Overview

Journal Mol Genet Genomics

Specialty Genetics

Date 2006 Jan 18

PMID 16416328

Citations 46

Authors

Kazuyuki Ojima

Jurgen Breitenbach

Hans Visser

Yutaka Setoguchi

Kazuyuki Tabata

Tatsuo Hoshino

Johan van den Berg

Gerhard Sandmann

Affiliations

Soon will be listed here.

Abstract

A gene has been cloned from Xanthophyllomyces dendrorhous by complementation of astaxanthin formation in a beta-carotene accumulating mutant. It consists of 3,166 bp and contains 17 introns. For the beta-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 beta-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 beta-carotene to astaxanthin formation by oxygenation of carbon 3 and 4. The reaction sequence is first 4-ketolation of beta-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 beta-ionone ends.

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