title: Purification, properties and induction of a specific benzoate-4-hydroxylase from Aspergillus niger (UBC 814).
abstract: An inducible benzoate-4-hydroxylase has been partially purified from crude extracts of the mycelial felts of Aspergillus niger. This enzyme catalyzes the transformation of benzoate to p-hydroxybenzoate with equimolar consumption of NADPH and O2. It requires tetrahydropteridine as a prosthetic group. The optimum activity was found at pH 6.2 with a Km value at 30 degrees C of 1.6-10-minus 4 for NADPH and 1.3-10-minus 4 M for benzoate. Fe-2+ (iron) is required for the enzyme activity. The enzyme is stabilized by the inclusion of benzoate, EDTA and glutathione in the extracting buffer. The enzyme is specific for benzoate as substrate. Sulfhydryl groups(s) are essential for enzyme activity as indicated by p-chloromercuri-benzoate and N-ethylmaleimide inactivation. Benzoate-4-hydroxylase activity is decreased in the mycelial felts of Aspergillus niger grown in the presence of higher concentrations of benzoate. Maximum activity of the enzyme was observed at 36 h after inoculation.
mesh_terms: D001230:Aspergillus / Q000201:enzymology*; D001565:Benzoates / Q000494:pharmacology; D002614:Chelating Agents / Q000494:pharmacology; D004790:Enzyme Induction; D006863:Hydrogen-Ion Concentration; D007501:Iron / Q000494:pharmacology; D007700:Kinetics; D006899:Mixed Function Oxygenases / Q000302:isolation & purification / Q000378:metabolism*; D009249:NADP / Q000494:pharmacology; D010101:Oxygen Consumption; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013696:Temperature; D013997:Time Factors
chemical_list: D001565:Benzoates; D002614:Chelating Agents; D013439:Sulfhydryl Reagents; D009249:NADP; D007501:Iron; D006899:Mixed Function Oxygenases