Characterization of an apo-carotenoid 13,14-dioxygenase from Novosphingobium aromaticivorans that converts β-apo-8'-carotenal to β-apo-13-carotenone.
A putative carotenoid oxygenase from Novosphingobium aromaticivorans was purified with a specific activity of 0.8 U/mg by His-Trap affinity chromatography. The native enzyme was estimated to be a 52 kDa monomer. Enzyme activity for β-apo-8'-carotenal was maximal at pH 8.0 and 45 °C, with a half life of 15.3 h, K(m) of 21 μM, and k(cat) of 25 l/min. The enzyme exhibited cleavage activity only for carotenoids containing one β-ionone ring and its catalytic efficiency (k(cat)/K(m)) followed the order β-apo-8'-carotenal > β-apo-4'-carotenal > γ-carotene. The enzyme converted these carotenoids to β-apo-13-carotenones by cleaving their C(13)-C(14) double bonds. The oxygen atom of β-apo-13-carotenone originated not from water but from molecular oxygen. Thus, the enzyme was an apo-carotenoid 13,14-dioxygenase.
Department of Bioscience and Biotechnology, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul 143-701, Republic of Korea.
Yeong-Su
YS
Kim
Eun-Sun
ES
Seo
Deok-Kun
DK
Oh
D002338:Carotenoids / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010105:Oxygenases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D042301:Sphingomonadaceae / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity; D013696:Temperature
D011994:Recombinant Proteins; C110652:beta-apo-8'-carotenal; C110027:beta-apocarotenoid-14',13'-dioxygenase; D002338:Carotenoids; C071310:beta-apo-13-carotenone; D010105:Oxygenases