RRC ID 19849
Author Kajiwara S, Kakizono T, Saito T, Kondo K, Ohtani T, Nishio N, Nagai S, Misawa N.
Title Isolation and functional identification of a novel cDNA for astaxanthin biosynthesis from Haematococcus pluvialis, and astaxanthin synthesis in Escherichia coli.
Journal Plant Mol. Biol.
Abstract We succeeded in isolating a novel cDNA involved in astaxanthin biosynthesis from the green alga Haematococcus pluvialis, by an expression cloning method using an Escherichia coli transformant as a host that synthesizes beta-carotene due to the Erwinia uredovora carotenoid biosynthesis genes. The cloned cDNA was shown to encode a novel enzyme, beta-carotene ketolase (beta-carotene oxygenase), which converted beta-carotene to canthaxanthin via echinenone, through chromatographic and spectroscopic analysis of the pigments accumulated in an E. coli transformant. This indicates that the encoded enzyme is responsible for the direct conversion of methylene to keto groups, a mechanism that usually requires two different enzymatic reactions proceeding via a hydroxy intermediate. Northern blot analysis showed that the mRNA was synthesized only in the cyst cells of H. pluvialis. E. coli carrying the H. pluvialis cDNA and the E. uredovora genes required for zeaxanthin biosynthesis was also found to synthesize astaxanthin (3S, 3'S), which was identified after purification by a variety of spectroscopic methods.
Volume 29(2)
Pages 343-52
Published 1995-10
PMID 7579184
MeSH Amino Acid Sequence Bacterial Proteins / genetics* Base Sequence Binding Sites / genetics Blotting, Northern Carotenoids / analysis Carotenoids / biosynthesis* Carotenoids / metabolism Chlorophyta / enzymology Chlorophyta / genetics* Chromatography, High Pressure Liquid Circular Dichroism Conserved Sequence DNA Mutational Analysis DNA, Complementary / genetics Escherichia coli / chemistry Escherichia coli / genetics Escherichia coli / metabolism Gene Library Molecular Sequence Data Oxygenases / genetics* Sequence Analysis, DNA Sequence Deletion Sequence Homology, Amino Acid Xanthophylls beta Carotene
IF 3.543
Times Cited 94
WOS Category PLANT SCIENCES BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Algae NIES-144