Characterization of Mycobacterium tuberculosis nicotinamidase/ pyrazinamidase

Hua Zhang, Jiao Yu Deng, Li Jun Bi, Ya Feng Zhou, Zhi Ping Zhang, Cheng Gang Zhang, Ying Zhang, Xian En Zhang

Research output: Contribution to journalArticlepeer-review

73 Scopus citations


The nicotinamidase/pyrazinamidase (PncA) of Mycobacterium tuberculosis is involved in the activation of the important front-line antituberculosis drug pyrazinamide by converting it into the active form, pyrazinoic acid. Mutations in the pncA gene cause pyrazinamide resistance in M. tuberculosis. The properties of M. tuberculosis PncA were characterized in this study. The enzyme was found to be a 20.89 kDa monomeric protein. The optimal pH and temperature of enzymatic activity were pH 7.0 and 40 °C, respectively. Inductively coupled plasma-optical emission spectrometry revealed that the enzyme was an Mn 2+/Fe2+-containing protein with a molar ratio of [Mn 2+] to [Fe2+] of 1 : 1; furthermore, the external addition of either type of metal ion had no apparent effect on the wild-type enzymatic activity. The activity of the purified enzyme was determined by HPLC, and it was shown that it possessed similar pyrazinamidase and nicotinamidase activity, by contrast with previous reports. Nine PncA mutants were generated by site-directed mutagenesis. Determination of the enzymatic activity and metal ion content suggested that Asp8, Lys96 and Cys138 were key residues for catalysis, and Asp49, His51, His57 and His71 were essential for metal ion binding. Our data show that M. tuberculosis PncA may bind metal ions in a manner different from that observed in the case of Pyrococcus horikoshii PncA.

Original languageEnglish (US)
Pages (from-to)753-762
Number of pages10
JournalFEBS Journal
Issue number4
StatePublished - Feb 2008


  • Mycobacterium tuberculosis
  • Nicotinamidase
  • PncA
  • Pyrazinamidase
  • Site-directed mutation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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