Preliminary report of NAD+-dependent amino acid dehydrogenase producing bacteria isolated from soil.
Amino acid dehydrogenases (L-amino acid: oxidoreductase deaminating; EC 1.4.1.X) are members of the wider superfamily of oxidoreductases that catalyze the reversible oxidative deamination of  an amino acid to its keto acid  and ammonia with the concomitant reduction of either NAD+, NADP+ or FAD. These enzymes have been received much  attention as biocatalysts for use in biosensors or diagnostic kits to screen amino acid metabolism disorders such as phenylketonuria (PKU), maple syrup urine disease (MSUD), homocystinuria (HCY) and hyperprolinemia. This study was aimed to isolation and screening of novel amino acid dehydrogenases from soil bacteriadehydrogenases from soil bacteria.



The enzyme producing bacteria were selected among L-methionine and L-phenylalanine utilizers isolated from soil by thin layer chromatography, activity staining and confirmed by enzyme assay. Bacterial strains were identified by phenotypic and biochemical characteristics. The steady-state kinetic studies of enzymes were also performed.



In total of 230 tested strains, four of them were recognized as amino acid dehydrogenase producers that belong to species of Pseudomonas, Citrobacter and Proteus. They exhibited the desired NAD+-dependent dehydrogenase activities toward L-isoleucine, L-methionine, L-cysteine, L-serine and L-glutamine in oxidative deamination reaction. The specific activity of L-isoleucine dehydrogenase, L-methionine dehydrogenase and L-glutamine dehydrogenase for oxidative deamination of L-isoleucine, L-methionine and L-glutamine were 1.59, 1.2 and 0.73 U/mg, respectively. The Kcat/Km (s(-1).mM(-1)) values in these strains were as follows: L-isoleucine, 113.6, L-methionine, 62.05 and L-glutamine, 95.83.



This is the first report of occurrence a specific isoleucine dehydrogenase, glutamine dehydrogenase and methionine dehydrogenase in bacteria.
Mohammadi
Hamid Shahbaz
HS
Young Researcher Club, Dept. of Biochemistry, Science & Research Campus, Islamic Azad University, Tehran, Iran.
Omidinia
Eskander
E
Dept.  of Biochemistry, Pasteur Institute of Iran, Tehran 13164, Iran.
Lotfi Sahebghadam
Abbas
A
Dept.  of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modarres University, Tehran  14115-115, Iran.
Saghiri
Reza
R
Dept.  of Biochemistry, Pasteur Institute of Iran, Tehran 13164, Iran.
D000594:Amino Acid Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism*; D000596:Amino Acids / Q000378:metabolism; D001419:Bacteria / Q000201:enzymology* / Q000302:isolation & purification; D006090:Gram-Negative Bacteria / Q000201:enzymology / Q000302:isolation & purification; D006094:Gram-Positive Bacteria / Q000201:enzymology / Q000302:isolation & purification; D007700:Kinetics; D009243:NAD / Q000378:metabolism*; D012988:Soil Microbiology
D000596:Amino Acids; D009243:NAD; D000594:Amino Acid Oxidoreductases