Tuberculosis is a world widespread disease, caused by Mycobacterium tuberculosis (M.tb). Although considered an obligate aerobe, this organism can resist life-limiting conditions such as microaerophily mainly due to its set of enzymes responsible for energy production and coenzyme restoration under these conditions. One of these enzymes is fumarate reductase, an heterotetrameric complex composed of a catalytic (FrdA), an iron-sulfur cluster (FrdB) and two transmembrane (FrdC and FrdD) subunits involved in anaerobic respiration and important for the maintenance of membrane potential. In this work, aiming to further characterize this enzyme function in mycobacteria, we analyzed the expression of FrdB-containing proteins in M.tb and Mycobacterium bovis Bacillus Calmette–Guérin (BCG) Moreau, the Brazilian vaccine strain against tuberculosis. We identified three isoforms in both mycobacteria, two of them corresponding to the predicted encoded polypeptides of M.tb (27 kDa) and BCG Moreau (40 kDa) frd sequences, as due to an insertion on the latter’s operon a fused FrdBC protein is expected. The third 52 kDa band can be explained by a transcriptional slippage event, typically occurring when mutation arises in a repetitive region within a coding sequence, thought to reduce its impact allowing the production of both native and variant forms. Comparative modeling of the M.tb and BCG Moreau predicted protein complexes allowed the detection of subtle overall differences, showing a high degree of structure and maybe functional resemblance among them. Axenic growth and macrophage infection assays show that the frd locus is important for proper bacterial development in both scenarios, and that both M.tb’s and BCG Moreau’s alleles can partially revert the hampered phenotype of the knockout strain. Altogether, our results show that the frdABCD operon of Mycobacteria may have evolved to possess other yet non-described functions, such as those necessary during aerobic logarithmic growth and early stage steps of infection.

Research on new fibrinolytic agents has been oriented towards the discovery of microbial proteases with plasmin-like activity, as alternative thrombolytic agents for thrombosis treatment. This study reports the characterization of an extracellular serine metalloprotease of approximately 56 kDa, with fibrinolytic and fibrinogenolytic activity, isolated from a S. marcescens strain obtained from the Amazon (CBAM 519). The enzyme showed optimum activity at pH 9 and temperature of 37 °C. Activity was reduced in the presence of Na+, Cu2+ and Fe2+ ions, and increased with Mn2⁺. The enzyme did not show hemolytic activity, nor did it activate plasminogen, but it effectively hydrolyzed fibrin and fibrinogen, rapidly degrading all fibrinogen chains Aα, Bβ and γ. Therefore, fibrinolysis was promoted only by the direct route, with a fast acting, potent fibrinolytic activity. Based on this, we expect the enzyme to be a protease of the plasmin type that directly degrades fibrin. These characteristics demonstrate great potential for its application in the treatment of thrombosis.

The Mycobacterium genus remains highly relevant today due to the rising incidence of tuberculosis and opportunistic infections caused by environmental mycobacteria. While much is known about M. tuberculosis, M. leprae and M. bovis, studies focusing on environmental mycobacteria remain limited. These microorganisms are globally distributed and have been identified in diverse biomes, including the Atlantic Forest. This study aims to provide a characterization of four mycobacterial strains isolated from the Atlantic Forest, assessing their metabolic capabilities and biotechnological potential. We investigated the presence of cellulases and proteases and conducted an initial profiling of secreted proteins. Furthermore, the examination of shared antigens and infection kinetics within macrophages offered insights into the ecological and pathogenic potential of these isolates. From a public health perspective, antigenic similarities between these environmental microorganisms and the BCG vaccine strain may influence the efficacy of BCG in protecting against diseases such as tuberculosis. Continued research on these and other environmental isolates, particularly within Brazil’s highly biodiverse ecosystems, holds promise for advancing scientific knowledge and contributing to human health.

Jatropha curcas contains seeds with a high oil content, suitable for biodiesel production. After oil extraction, the remaining mass can be a rich source of enzymes. However, data from the literature describing physicochemical characteristics for a monomeric esterase from the J. curcas seed did not fit the electrostatic catapult model for esterases/lipases. We decided to reevaluate this J. curcas esterase and extend its characterization to check this apparent discrepancy and gain insights into the enzyme’s potential as a biocatalyst. After anion exchange chromatography and two-dimensional gel electrophoresis, we identified the enzyme as belonging to the dienelactone hydrolase family, characterized by a cysteine as the nucleophile in the catalytic triad. The enzyme displayed a basic optimum hydrolysis pH of 9.0 and an acidic pI range, in contrast to literature data, making it well in line with the electrostatic catapult model. Furthermore, the enzyme showed low hydrolysis activity in an organic solvent-containing medium (isopropanol, acetonitrile, and ethanol), which reverted when recovering in an aqueous reaction mixture. This enzyme can be a valuable tool for hydrolysis reactions of short-chain esters, useful for pharmaceutical intermediates synthesis, due to both its high hydrolytic rate in basic pH and its stability in an organic solvent.