The protective effect of a novel antioxidant gene from Mycobacterium avium against nitrosative and oxidative stress...

The production of reactive oxygen intermediates (ROI) and reactive nitrogen intermediates (RNI) is an important host defense mechanism in response to infection by Mycobacterium tuberculosis. A variety of genes have been implicated in resistance to ROI and RNI, including noxR1. However, studies in Mycobacterium avium, an important pathogen among nontuberculous mycobacteria, are limited. We aim to investigate the role of a novel gene cloned from M. avium with high similarity to noxR1, noA, in resistance against RNI and ROI in M. tuberculosis. After subcloning noA into vector for expression in E. coli, we performed survival rate analysis in the bacteria transformed with noA (pET-noA) and without noA (pET-his) after exposure to nitrosative stresses by S-nitrosoglutathione (GSNO) and sodium nitrite, and oxidative stresses by H₂O₂. Compared with pET-his, the survival rate of pET-noA was 1 log₁₀-fold higher after exposure to GSNO and sodium nitrite. We observed 1 log₁₀-fold, 2 log₁₀-fold and 3 log₁₀-fold higher survival rate in pET-noA than pET-his after exposure to H₂O₂ for 3, 6 and 9 h, respectively. With the combined treatment of H₂O₂ and GSNO, we found more than 2 log₁₀-fold increase in survival rate in pET-noA comparing with pET-his, suggesting a possible synergistic effect. In summary, noA gene cloned from M. avium has been shown to protect E. coli from both RNI and ROI.