Purification and characterization of an extracellular adenosine deaminase from Nocardioides sp. J-326TK

Biotechnol Appl Biochem. 1994 Oct;20(2):265-77.


An extracellular adenosine deaminase was isolated from the culture supernatant of Nocardioides sp. J-326TK and purified 193-fold to homogeneity. It had a specific activity of 4677 units/mg at 37 degrees C, was a monomeric protein as judged by SDS/PAGE, and was characterized with respect to M(r) (80,000 and 72,000 by gel filtration on Sephadex G-200 and SDS/PAGE respectively), pH optimum (6.0), temperature optimum (50 degrees C) and pI (7.6). The adsorption spectrum of the enzyme had a maximum at 280 nm and a minimum at 250 nm. The enzyme was stable at pH 6.5-7.5 and at temperatures below 30 degrees C. Adenosine and 2'-deoxyadenosine were deaminated and the respective Km values were 0.22 and 0.20 mM, but the enzyme was not active on adenine and 6-(gamma gamma'-dimethylallylamino)purine riboside. The enzyme reaction was promoted by Fe3+ and Sn2+, but potently inhibited by Hg2+, Ag2+, o-phenanthroline and pentachlorophenol, and noticeably inhibited by 8-bromoadenosine, theobromine and theophylline.

MeSH terms

  • Adenosine Deaminase / biosynthesis
  • Adenosine Deaminase / drug effects
  • Adenosine Deaminase / isolation & purification*
  • Culture Media
  • Enzyme Stability
  • Hydrogen-Ion Concentration
  • Isoenzymes / biosynthesis
  • Isoenzymes / drug effects
  • Isoenzymes / isolation & purification*
  • Metals / pharmacology
  • Molecular Weight
  • Nocardiaceae / enzymology*
  • Nucleosides / chemistry
  • Spectrophotometry, Ultraviolet
  • Substrate Specificity
  • Temperature


  • Culture Media
  • Isoenzymes
  • Metals
  • Nucleosides
  • Adenosine Deaminase