NAD deamidation "a new reaction" by an enzyme from Aspergillus terreus DSM 826.
NAD deamidation is a non-previously recognized reaction. This reaction has been found to be catalyzed by extracts of Aspergillus terreus DSM 826. Conversion of NAD to the biosynthetic intermediate, deamido NAD, by these extracts, at the optimum pH and temperature did not exceed about 55 of the amount of the substrate added. Completion of the reaction was achieved when the extracts were pre-heated at 50 degrees C for 15 min in absence of the substrate. In a very similar manner, the extracts catalyzed hydrolytic cleavage of the amide linkages of different biomolecules such as nicotinamide, nicotinamide riboside, nicotinamide mononucleotide, L-glutamine, L-asparagine and acetamide. Polyacrylamide was also deamidated under the same conditions. In addition, complete dephosphorylation of the dinucleotide molecule was also effected by the same extracts. Separation of the NAD deamidating enzyme from the NAD dephosphorylating enzyme was achieved on using either DEAE - Sephadex A-25 or Sephadex G-200 column chromatography. The obtained phosphohydrolase-free-deamidase showed optimum activity at pH 8 of 0.1 M phosphate buffer and 50 degrees C. It exhibited broad substrate specificity and hyperbolic substrate saturation kinetics. It was isosterically inhibited by the product of its activity and this inhibition was prevented by heating the extracts at 50 degrees C for 15 min. Its activity was not affected in presence of sodium fluoride, partially inhibited in presence of magnesium chloride and was retained in the freezer for some months.
Elzainy
Tahany A
TA
Department of Microbial Chemistry, National Research Centre, Dokki, Cairo, Egypt. ems@internetegypt.com
Ali
Thanaa H
TH
D000081:Acetamides / Q000378:metabolism; D000180:Acrylic Resins / Q000378:metabolism; D001216:Asparagine / Q000378:metabolism; D001230:Aspergillus / Q000201:enzymology* / Q000378:metabolism; D002853:Chromatography, Liquid; D003641:Deamination; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability; D005615:Freezing; D005973:Glutamine / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009243:NAD / Q000031:analogs & derivatives* / Q000378:metabolism*; D009536:Niacinamide / Q000031:analogs & derivatives* / Q000378:metabolism; D009537:Nicotinamide Mononucleotide / Q000378:metabolism; D011726:Pyridinium Compounds; D013379:Substrate Specificity; D013696:Temperature; D000099166:Polyacrylamides
D000081:Acetamides; D000180:Acrylic Resins; D001216:Asparagine; D004791:Enzyme Inhibitors; D005973:Glutamine; D009243:NAD; D009536:Niacinamide; D009537:Nicotinamide Mononucleotide; D011726:Pyridinium Compounds; D000099166:Polyacrylamides; C018613:nicotinamide-beta-riboside; C018348:nicotinic acid adenine dinucleotide; C030686:acetamide