Solubilization and reconstitution of the membrane-bound carotenogenic enzymes from daffodil chromoplasts.
The membrane-bound carotenogenic enzymes of daffodil (Narcissus pseudonarcissus) chromoplast membranes, i.e. dehydrogenation, cis-trans isomerization and cyclization activities, were solubilized in an enzymatically inactive form using the zwitterionic detergent 3-[(3-cholamidopropyl)-dimethylamino]-1-propanesulfonate (Chaps). Full enzymatic activities were regained upon reconstitution of the solubilized proteins into liposomes. These preparations converted radiolabelled cis-phytoene into beta-carotene in a high yield. The reconstituted enzymatic sequence behaved as a tight 'assembly line'. In the enzymatic sequence of the reconstituted system the cis-trans isomerization reaction occurred on the stage of cis-phytofluene after a dehydrogenation of cis-phytoene.
D001711:Biotransformation; D002338:Carotenoids / Q000096:biosynthesis*; D002462:Cell Membrane / Q000201:enzymology; D003902:Detergents; D008081:Liposomes / Q000378:metabolism; D008854:Microscopy, Electron; D009940:Organoids / Q000201:enzymology; D010944:Plants / Q000201:enzymology*; D011485:Protein Binding; D012995:Solubility
D003902:Detergents; D008081:Liposomes; D002338:Carotenoids