Cloning and expression of beta,beta-carotene 15,15'-dioxygenase.
beta,beta-Carotene 15,15'-dioxygenase cleaves beta-carotene into two molecules of retinal and is therefore the key enzyme in beta-carotene metabolism to vitamin A. In the present study, it was possible to enrich the chicken beta,beta-carotene 15,15'-dioxygenase to such an extent that partial amino acid sequence information could be obtained to design degenerate oligonucleotides. With RT-PCR a cDNA fragment could be obtained and used subsequently in a radioactive screening of a chicken duodenal expression library. We cloned the first eukaryotic beta,beta-carotene 15,15'-dioxygenase which symmetrically cleaves beta-carotene at the 15,15'-double bond.
Wyss
A
A
Vitamins & Fine Chemicals Division, F. Hoffmann-LaRoche Ltd., Basel, 4070, Switzerland. adrian.wyss@roche.com
Wirtz
G
G
Woggon
W
W
Brugger
R
R
Wyss
M
M
Friedlein
A
A
Bachmann
H
H
Hunziker
W
W
D000818:Animals; D002460:Cell Line; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D006224:Cricetinae; D003593:Cytoplasm / Q000201:enzymology; D018076:DNA, Complementary / Q000378:metabolism; D004386:Duodenum / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D010105:Oxygenases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry; D011994:Recombinant Proteins; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D050559:beta-Carotene 15,15'-Monooxygenase
D018076:DNA, Complementary; D010455:Peptides; D011994:Recombinant Proteins; D010105:Oxygenases; C493359:Bco1 protein, mouse; D050559:beta-Carotene 15,15'-Monooxygenase