title: CYP175A1 from Thermus thermophilus HB27, the first beta-carotene hydroxylase of the P450 superfamily.
abstract: The biological function of thermostable P450 monooxygenase CYP175A1 from Thermus thermophilus HB27 was studied by functional complementation in Escherichia coli. The gene product of CYP175A1 added hydroxyl groups to both beta rings of beta-carotene to form zeaxanthin (beta,beta-carotene-3,3'-diol) in E. coli, which produces beta-carotene due to the Erwinia uredovora carotenoid biosynthesis genes. In addition, spectroscopic methods revealed that E. coli carrying CYP175A1 and the cDNA of the Haematococcus pluvialis carotene ketolase was able to synthesise hydroxyechinenone. The predicted amino acid sequence of the enzyme from T. thermophilus does not show substantial similarity with other known beta-carotene hydroxylases, but 41% with the cytochrome P450 monooxygenase from Bacillus megaterium (CYP102A1, P450 BM3). It is concluded that CYP175 A1 represents a new type of beta-carotene hydroxylase of the P450 superfamily.
authors.affiliation: Institute of Technical Biochemistry, University of Stuttgart, 70569, Stuttgart, Germany.
authors.forename: F
authors.initials: F
authors.lastname: Blasco
authors.forename: I
authors.initials: I
authors.lastname: Kauffmann
authors.forename: R D
authors.initials: RD
authors.lastname: Schmid
mesh_terms: D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004885:Erwinia / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005816:Genetic Complementation Test; D007527:Isoenzymes; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism*; D016963:Thermus thermophilus / Q000201:enzymology* / Q000235:genetics; D014170:Transformation, Genetic / Q000235:genetics; D019207:beta Carotene / Q000378:metabolism*
chemical_list: D004269:DNA, Bacterial; D007527:Isoenzymes; D019207:beta Carotene; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; C079232:beta-carotene hydroxylase