Cloning and expression of beta,beta-carotene 15,15'-dioxygenase

Biochem Biophys Res Commun. 2000 May 10;271(2):334-6. doi: 10.1006/bbrc.2000.2619.

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.

MeSH terms

  • Animals
  • Cell Line
  • Chickens
  • Chromatography, High Pressure Liquid
  • Cloning, Molecular
  • Cricetinae
  • Cytoplasm / enzymology
  • DNA, Complementary / metabolism
  • Duodenum / enzymology
  • Electrophoresis, Polyacrylamide Gel
  • Oxygenases / chemistry
  • Oxygenases / genetics*
  • Oxygenases / metabolism*
  • Peptides / chemistry
  • Recombinant Proteins
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • beta-Carotene 15,15'-Monooxygenase

Substances

  • DNA, Complementary
  • Peptides
  • Recombinant Proteins
  • Oxygenases
  • Bco1 protein, mouse
  • beta-Carotene 15,15'-Monooxygenase