Amino Acids

, Volume 38, Issue 1, pp 247–255 | Cite as

d-Amino acid dehydrogenase from Helicobacter pylori NCTC 11637

  • Minoru Tanigawa
  • Tomomitsu Shinohara
  • Makoto Saito
  • Katsushi Nishimura
  • Yuichiro Hasegawa
  • Sadao Wakabayashi
  • Morio Ishizuka
  • Yoko Nagata
Original Article


Helicobacter pylori is a microaerophilic bacterium, associated with gastric inflammation and peptic ulcers. d-Amino acid dehydrogenase is a flavoenzyme that digests free neutral d-amino acids yielding corresponding 2-oxo acids and hydrogen. We sequenced the H. pylori NCTC 11637 d-amino acid dehydrogenase gene, dadA. The primary structure deduced from the gene showed low similarity with other bacterial d-amino acid dehydrogenases. We purified the enzyme to homogeneity from recombinant Escherichia coli cells by cloning dadA. The recombinant protein, DadA, with 44 kDa molecular mass, possessed FAD as cofactor, and showed the highest activity to d-proline. The enzyme mediated electron transport from d-proline to coenzyme Q1, thus distinguishing it from d-amino acid oxidase. The apparent Km and Vmax values were 40.2 mM and 25.0 μmol min−1 mg−1, respectively, for dehydrogenation of d-proline, and were 8.2 μM and 12.3 μmol min−1 mg−1, respectively, for reduction of Q1. The respective pH and temperature optima were 8.0 and 37°C. Enzyme activity was inhibited markedly by benzoate, and moderately by SH reagents. DadA showed more similarity with mammalian d-amino acid oxidase than other bacterial d-amino acid dehydrogenases in some enzymatic characteristics. Electron transport from d-proline to a c-type cytochrome was suggested spectrophotometrically.


d-Amino acid dehydrogenase d-Proline Gene cloning Helicobacter pylori Bacterial respiration 



d-Amino acid dehydrogenase


DAD purified from recombinant Escherichia coli




Phenazine methosulfate


Sodium dodecyl sulfate-polyacrylamide gel electrophoresis



We would like to express our thanks to Dr. K. Nagata (Department of Bacteriology, Hyogo College of Medicine) for providing H. pylori NCTC 11637 strain and useful advice for culture.


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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Minoru Tanigawa
    • 1
  • Tomomitsu Shinohara
    • 1
  • Makoto Saito
    • 1
  • Katsushi Nishimura
    • 1
    • 2
  • Yuichiro Hasegawa
    • 1
  • Sadao Wakabayashi
    • 3
  • Morio Ishizuka
    • 4
  • Yoko Nagata
    • 1
  1. 1.Department of Materials and Applied Chemistry, College of Science and TechnologyNihon UniversityTokyoJapan
  2. 2.Department of Applied Chemistry, Junior CollegeNihon UniversityChibaJapan
  3. 3.Department of Life Sciences, Graduate School of Life ScienceUniversity of HyogoHyogoJapan
  4. 4.Department of Applied Chemistry, Faculty of Science and EngineeringChuo UniversityTokyoJapan

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