Cloning and expression of beta,beta-carotene 15,15'-dioxygenase |
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Authors: | Wyss A Wirtz G Woggon W Brugger R Wyss M Friedlein A Bachmann H Hunziker W |
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Affiliation: | Vitamins & Fine Chemicals Division, F. Hoffmann-LaRoche Ltd., Basel, 4070, Switzerland. adrian.wyss@roche.com |
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Abstract: | 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. |
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