A putative oxidoreductase, Rv1714 from Mycobacterium tuberculosis, predicted as a short-chain...

Oxidoreductases are a class of enzymes that transfer electrons in various biological pathways and reactions, at the expense of cellular NADPH/NADH. Because oxidoreductases are involved in numerous energy metabolism pathways, many of these enzymes have been identified as potential drug targets. Although oxidoreductases have been implicated in energy metabolism pathways and are crucial for the survival and pathogenicity of Mycobacterium tuberculosis (M.tb), their comprehensive characterization remains limited. In the present study, one uncharacterized and putative oxidoreductase, Rv1714, was identified in the H37Rv proteome. This putative oxidoreductase was conserved across pathogenic and opportunistic mycobacterial species and was predicted to be a virulence factor essential for the pathogenicity. Rv1714 displayed homology with the short-chain dehydrogenase/reductase (SDR) family. The recombinant Rv1714 was purified to homogeneity, and its structural properties were explored using CD and fluorescence spectroscopies, as well as bioinformatics tools. The sequence and structural homologies revealed the characteristic features and conservation of the active site with the SDR family of proteins. Possible substrates were explored using docking, simulation, and MMPBSA calculations. Furthermore, in vitro studies using macrophage cell lines demonstrate that recombinant Rv1714 significantly enhances the production of pro-inflammatory cytokines, including IL1β, TNF-α, and IL-6, suggesting that Rv1714 may play a critical role in modulating the host's innate immune response during infection. The stimulation of these cytokines is associated with the activation of the NF-κB pathway. Altogether, this is the first report on the putative oxidoreductase Rv1714 of M.tb, highlighting its structural features and its key immune-modulatory function involved in shaping host inflammatory responses during infection.