This research was funded by FONDECYT-CONCYTEC (grant contract number 271-2015-FONDECYT ), the São Paulo Research Foundation (FAPESP, Brazil) for funding the project N° 2018/05871-3 and CNPq proc . Num. 305295/2018-7 .

Erick Saldaña received the support of the “Consejo Nacional de Ciencia, Tecnología e Innovación Tecnológica - CONCYTEC" from Perú (CIENCIACTIVA programme, PhD scholarship contract agreement No. 104-2016-FONDECYT).

Mycobacterium tuberculosis nicotinamidase-pyrazinamidase (PZAse) is a metalloenzyme that catalyzes conversion of nicotinamide-pyrazinamide to nicotinic acid-pyrazinoic acid. This study investigated whether a metallochaperone is required for optimal PZAse activity. M. tuberculosis and Escherichia coli PZAses (PZAse-MT and PZAse-EC, respectively) were inactivated by metal depletion (giving PZAse-MT–Apo and PZAse-EC–Apo). Reactivation with the E. coli metallochaperone ZnuA or Rv2059 (the M. tuberculosis analog) was measured. This was repeated following proteolytic and thermal treatment of ZnuA and Rv2059. The CDC1551 M. tuberculosis reference strain had the Rv2059 coding gene knocked out, and PZA susceptibility and the pyrazinoic acid (POA) efflux rate were measured. ZnuA (200 M) achieved 65% PZAse-EC–Apo reactivation. Rv2059 (1 M) and ZnuA (1 M) achieved 69% and 34.3% PZAse-MT–Apo ...