Cloning, overexpression and characterization of Leishmania donovani triosephosphate isomerase

Structural insights from a novel invertebrate triosephosphate isomerase from Litopenaeus vannamei

“…3B ). The optimal pH is in agreement with reports by Kumar et al where the L. donovani is more stable at pH 8.8, ranging from 6.5-8.5 [18]. Although enzyme optimal temperature may or not have a correlation with the environment of the organisms, as shrimp does not regulate its body temperature, the structural stability to unfolding will provide more insights into the structure and function relationship.…”

“…3A ). The K m of LvTIM was 0.7 mM, higher than the K m reported for human TIM (0.49 mM) [5] and TIMs from protozoan parasites such as L. donovani (0.328 mM) [18], L. mexicana (0.30 mM) [37], G. lamblia (0.53 mM) [38], T. brucei (0.25 mM) [39], but lower than the K m from TIMs from Vibrio marinus (1.9 mM) [9] or S. cerevisiae (1.27 mM) [40]. The LvTIM k cat was 1.2 × 10 5 min −1 , similar to the k cat reported for other TIMs such L. mexicana (2.5 ×10 5 min −1 ) [37], Helicobacter pylori (8.8 ×10 4 min −1 ) [41], T. brucei (3.7 ×10 5 min −1 ) [39] and rabbit muscle TIM (5.1 ×10 5 min −1 ) [42].…”

“…The activity assay was done as previously reported [18], following the decrease in absorbance at 340 nm at 25 °C due to the oxidation of NADH. The assay was done in a 1 ml quartz cell containing 50 mM Tris pH 7.6, 1 mM glyceraldehyde 3-phosphate, 0.2 mM NADH and 1 U of α-glycerophosphate dehydrogenase [29].…”

“…The TIM dimer interface is a starting point towards rational drug design against diseases originated by protozoan parasites that heavily depend on glycolysis for energy [15]. Molecules that interact with exposed cysteines like methylmethane thiosulfonate (MMTS) selectively inhibit TIMs from Leishmania donovani [18], T. cruzi [19], T. brucei [20] and Giardia lamblia [21]. The strength of the inhibition is dependent on the degree of conjugation of a reactive cysteine (Cys15 in the case of T. cruzi ) and the perturbation of the interactions at the homodimeric interface.…”

Gene Cloning, Recombinant Expression, Characterization, and Molecular Modeling of the Glycolytic Enzyme Triosephosphate Isomerase from Fusarium oxysporum

“…The main function of the TPI enzyme is to catalyze the interconversion of dihydroxyacetone phosphate into (D)-glyceraldehyde-3-phosphate; this process allows the two three-carbon molecules to continue being processed in the glycolytic pathway, since, without this reaction, there would be no net ATP production generated by the pathway [19]. Due to the relevance of this enzyme, some researchers have proposed the TPI as a potential target for the search (or synthesis) for new drugs [15][16][17][18], and even propose it as targets for the development of vaccines, of parasites that affect humans [16,20,21]. According to the aforementioned, it was in our interest to study the TPI of the fungus identified as F. oxysporum, since, despite being a pathogenic fungus of plants and/or humans, the analysis of biochemical characterization of this or another enzyme of the glycolysis had not been performed previously.…”