Cloning and expression inEscherichia coli of the gene encoding β-C-4-oxygenase, that converts β-carotene to the ketocarotenoid canthaxanthin inHaematococcus pluvialis

Tamar Lotan; Joseph Hirschberg

doi:10.1016/0014-5793(95)00368-J

Cloning and expression inEscherichia coli of the gene encoding β-C-4-oxygenase, that converts β-carotene to the ketocarotenoid canthaxanthin inHaematococcus pluvialis

Tamar Lotan, Joseph Hirschberg

Research output: Contribution to journal › Article › peer-review

Abstract

In the green algaHaematococcus pluvialis the ketocarotenoid astaxanthin accumulates under stress conditions. Astaxanthin is a red carotenoid pigment which possess antioxidative activity. We have cloned the gene for β-C-4 oxygenase (β-carotene ketolase) from the green algaeH. pluvialis. The cloning method took advantage of a strain ofE. coli which was genetically engineered to produce β-carotene. An expression cDNA library ofH. pluvialis was transfected to cells of this strain and visually screened for brown-red pigmented colonies. One colony out of 100,000 transformants showed color change due to accumulation of canthaxanthin. The cDNA clone in this transformant colony encodes the enzyme β-C-4 oxygenase that catalyzes the conversion of β carotene to canthaxanthin via echinenone. This enzyme does not convert zeaxanthin to astaxanthin. It is concluded that inH. pluvialis astaxanthin is synthesized via canthaxanthin and therefore an additional enzyme is predicted, which converts canthaxanthin to astaxanthin.

Original language	English
Pages (from-to)	125-128
Number of pages	4
Journal	FEBS Letters
Volume	364
Issue number	2
DOIs	https://doi.org/10.1016/0014-5793(95)00368-J
State	Published - 8 May 1995
Externally published	Yes

Bibliographical note

Funding Information:
Acknowledgments: This research was supported by Grant 034-7342 from the Ernest D. Bergman Fund for Science. T. Lotan is supported by the L. Eshkol grant of the Ministry of Science, National Council for Research and Development. We wish to thank Dr. A. Young for supplying the carotenoid standards and Dr. A. Mandelbaum and Dr. J. Katzir for the mass spectrometry analysis.

Keywords

Astaxanthin
Biosynthesis
Canthaxanthin
Carotenoid
Cloning
Haematococcus pluvialis

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1016/0014-5793(95)00368-J

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title = "Cloning and expression inEscherichia coli of the gene encoding β-C-4-oxygenase, that converts β-carotene to the ketocarotenoid canthaxanthin inHaematococcus pluvialis",

abstract = "In the green algaHaematococcus pluvialis the ketocarotenoid astaxanthin accumulates under stress conditions. Astaxanthin is a red carotenoid pigment which possess antioxidative activity. We have cloned the gene for β-C-4 oxygenase (β-carotene ketolase) from the green algaeH. pluvialis. The cloning method took advantage of a strain ofE. coli which was genetically engineered to produce β-carotene. An expression cDNA library ofH. pluvialis was transfected to cells of this strain and visually screened for brown-red pigmented colonies. One colony out of 100,000 transformants showed color change due to accumulation of canthaxanthin. The cDNA clone in this transformant colony encodes the enzyme β-C-4 oxygenase that catalyzes the conversion of β carotene to canthaxanthin via echinenone. This enzyme does not convert zeaxanthin to astaxanthin. It is concluded that inH. pluvialis astaxanthin is synthesized via canthaxanthin and therefore an additional enzyme is predicted, which converts canthaxanthin to astaxanthin.",

keywords = "Astaxanthin, Biosynthesis, Canthaxanthin, Carotenoid, Cloning, Haematococcus pluvialis",

author = "Tamar Lotan and Joseph Hirschberg",

note = "Funding Information: Acknowledgments: This research was supported by Grant 034-7342 from the Ernest D. Bergman Fund for Science. T. Lotan is supported by the L. Eshkol grant of the Ministry of Science, National Council for Research and Development. We wish to thank Dr. A. Young for supplying the carotenoid standards and Dr. A. Mandelbaum and Dr. J. Katzir for the mass spectrometry analysis.",

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AU - Lotan, Tamar

AU - Hirschberg, Joseph

N1 - Funding Information: Acknowledgments: This research was supported by Grant 034-7342 from the Ernest D. Bergman Fund for Science. T. Lotan is supported by the L. Eshkol grant of the Ministry of Science, National Council for Research and Development. We wish to thank Dr. A. Young for supplying the carotenoid standards and Dr. A. Mandelbaum and Dr. J. Katzir for the mass spectrometry analysis.

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Y1 - 1995/5/8

N2 - In the green algaHaematococcus pluvialis the ketocarotenoid astaxanthin accumulates under stress conditions. Astaxanthin is a red carotenoid pigment which possess antioxidative activity. We have cloned the gene for β-C-4 oxygenase (β-carotene ketolase) from the green algaeH. pluvialis. The cloning method took advantage of a strain ofE. coli which was genetically engineered to produce β-carotene. An expression cDNA library ofH. pluvialis was transfected to cells of this strain and visually screened for brown-red pigmented colonies. One colony out of 100,000 transformants showed color change due to accumulation of canthaxanthin. The cDNA clone in this transformant colony encodes the enzyme β-C-4 oxygenase that catalyzes the conversion of β carotene to canthaxanthin via echinenone. This enzyme does not convert zeaxanthin to astaxanthin. It is concluded that inH. pluvialis astaxanthin is synthesized via canthaxanthin and therefore an additional enzyme is predicted, which converts canthaxanthin to astaxanthin.

AB - In the green algaHaematococcus pluvialis the ketocarotenoid astaxanthin accumulates under stress conditions. Astaxanthin is a red carotenoid pigment which possess antioxidative activity. We have cloned the gene for β-C-4 oxygenase (β-carotene ketolase) from the green algaeH. pluvialis. The cloning method took advantage of a strain ofE. coli which was genetically engineered to produce β-carotene. An expression cDNA library ofH. pluvialis was transfected to cells of this strain and visually screened for brown-red pigmented colonies. One colony out of 100,000 transformants showed color change due to accumulation of canthaxanthin. The cDNA clone in this transformant colony encodes the enzyme β-C-4 oxygenase that catalyzes the conversion of β carotene to canthaxanthin via echinenone. This enzyme does not convert zeaxanthin to astaxanthin. It is concluded that inH. pluvialis astaxanthin is synthesized via canthaxanthin and therefore an additional enzyme is predicted, which converts canthaxanthin to astaxanthin.

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KW - Carotenoid

KW - Cloning

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Cloning and expression inEscherichia coli of the gene encoding β-C-4-oxygenase, that converts β-carotene to the ketocarotenoid canthaxanthin inHaematococcus pluvialis

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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