title: Purification and partial characterization of NAD aminohydrolase from Aspergillus oryzae NRRL447.
abstract: Aspergillus oryzae aminohydrolase free acid phosphodiesterase catalyzes nicotinamide adenine dinucleotide to deamino-NAD and ammonia. The enzyme was purified to homogeneity by a combination of acetone precipitation, anion exchange chromatography and gel filtration chromatography. The enzyme was purified 230.5 fold. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme showed a single protein band of MW 94 kDa. The enzyme displayed maximum activity at pH 5 and 40 °C with NAD as substrate. The enzyme activity appeared to be stable up to 40 °C. The enzyme activity was enhanced slightly by addition of Na⁺ and K⁺, whereas inhibited strongly by addition of Ag⁺, Mn²⁺, Hg²⁺ and Cu²⁺ to the reaction mixtures. The enzyme hydrolyzes several substrates, suggesting a probable non-specific nature. The enzyme catalyzes the hydrolytic cleavage of amino group of NAD, adenosine, AMP, CMP, GMP, adenosine, cytidine and cytosine to the corresponding nucleotides, nucleosides or bases and ammonia. The substrate concentration-activity relationship is the hyperbolic type and the apparent Km and Kcat for the tested substrates were calculated.
authors.affiliation: Department of Microbial Chemistry, National Research Centre, Dokki, Cairo, Egypt, thanaa15@yahoo.com.
authors.forename: Thanaa H
authors.initials: TH
authors.lastname: Ali
authors.forename: Nadia H
authors.initials: NH
authors.lastname: Ali
authors.forename: Bakry M
authors.initials: BM
authors.lastname: Haroun
authors.forename: Amir E
authors.initials: AE
authors.lastname: Tantawy
mesh_terms: D000619:Aminohydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D000641:Ammonia / Q000378:metabolism; D001236:Aspergillus oryzae / Q000201:enzymology*; D011232:Chemical Precipitation; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003067:Coenzymes / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D020536:Enzyme Activators / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000378:metabolism; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008670:Metals / Q000378:metabolism; D008970:Molecular Weight; D009243:NAD / Q000031:analogs & derivatives / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D003067:Coenzymes; D020536:Enzyme Activators; D004791:Enzyme Inhibitors; D008670:Metals; D009243:NAD; C032140:nicotinamide-hypoxanthine dinucleotide; D000641:Ammonia; D000619:Aminohydrolases