3,4-Dihydroxyxanthone dioxygenase from Arthrobacter sp. strain GFB100.
Bacterial extradiol ring-fission dioxygenases play a critical role in the transformation of multiring aromatic compounds to more readily biodegradable aromatic or aliphatic intermediates. Arthrobacter sp. strain GFB100 utilizes an extradiol meta-fission dioxygenase, 3,4-dihydroxyxanthone dioxygenase (DHXD), in the catabolism of the three-ring oxygen heterocyclic compound xanthone. In this paper, we show that DHXD is a cytosolic enzyme, induced by growth on xanthone and maximally expressed during the stationary phase of growth. In addition, we characterize the DHXD activity in terms of its basic enzymological properties. 1,10-Phenanthroline and H2O2 treatments eliminated DHXD activity, indicating that the enzyme required Fe2+ ions for activity. Other divalent cations were either inhibitory or had no effect on activity. DHXD had a temperature optimum of 30 degrees C and a pH optimum of 7.0. DHXD followed typical saturation kinetics and had an apparent Km of 10 microM for 3,4-dihydroxyxanthone. The dye celestine blue served as a noncompetitive DHXD inhibitor (Ki, 5 microM). Several other structural analogs served neither as substrates nor inhibitors. DHXD was thermally labile at temperatures above 40 degrees C. The half-life for thermal DHXD inactivation was 5 min at 40 degrees C. DHXD activity was completely stable through one freeze-thaw cycle, and about 80% of the DHXD activity remained after 2 days of incubation at 0 degree C. The apparent tight binding of the Fe2+ cofactor to DHXD may be a factor contributing to the stability of this extradiol dioxygenase when it is stored.
Chen
C M
CM
Department of Food Science, Cook College, New Jersey Agricultural Experiment Station, Rutgers University, New Brunswick 08903.
Tomasek
P H
PH
D001173:Arthrobacter / Q000201:enzymology* / Q000254:growth & development; D049308:Dioxygenases; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007502:Iron Chelating Agents / Q000494:pharmacology; D007700:Kinetics; D010105:Oxygenases / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature; D014966:Xanthenes / Q000378:metabolism; D044004:Xanthones
D007502:Iron Chelating Agents; D014966:Xanthenes; D044004:Xanthones; C009689:xanthone; D010105:Oxygenases; C072255:3,4-dihydroxyxanthone dioxygenase; D049308:Dioxygenases