Expression and characterization of a β-fructofuranosidase from the parasitic protist Trichomonas vaginalis

Abstract

BackgroundTrichomonas vaginalis, a flagellated protozoan, is the agent responsible for trichomoniasis, the most common nonviral sexually transmitted infection worldwide. A reported 200 million cases are documented each year with far more cases going unreported. However, T. vaginalis is disproportionality under studied, especially considering its basic metabolism. It has been reported that T. vaginalis does not grow on sucrose. Nevertheless, the T. vaginalis genome contains some 11 putative sucrose transporters and a putative β-fructofuranosidase (invertase). Thus, the machinery for both uptake and cleavage of sucrose appears to be present.ResultsWe amplified the β-fructofuranosidase from T. vaginalis cDNA and cloned it into an Escherichia coli expression system. The expressed, purified protein was found to behave similarly to other known β-fructofuranosidases. The enzyme exhibited maximum activity at pH close to 5.0, with activity falling off rapidly at increased or decreased pH. It had a similar Km and Vmax to previously characterized enzymes using sucrose as a substrate, was also active towards raffinose, but had no detectable activity towards inulin.ConclusionsT. vaginalis has the coding capacity to produce an active β-fructofuranosidase capable of hydrolyzing di- and trisaccharides containing a terminal, non-reducing fructose residue. Since we cloned this enzyme from cDNA, we know that the gene in question is transcribed. Furthermore, we could detect β-fructofuranosidase activity in T. vaginalis cell lysates. Therefore, the inability of the organism to utilize sucrose as a carbon source cannot be explained by an inability to degrade sucrose.

Promising Prebiotic Candidate Established by Evaluation of Lactitol, Lactulose, Raffinose, and Oligofructose for Maintenance of a Lactobacillus-Dominated Vaginal Microbiota

“…This is further supported by the lack of fermentation products produced and the high levels of raffinose remaining in the spent media of the vaginal sample. However, other researchers have observed that raffinose can be used by Trichomonas vaginalis ( 43 ), the vaginal parasite responsible for trichomoniasis, which further underscores the unsuitability of this compound as an effective vaginal health agent.…”

Investigation of Antioxidant, Antifungal, Antibacterial, and Anti-inflammatory Effects of Teucrium polium on Common Pathogens in Vaginitis: A Review

“…To treat this infection, various antibiotics, of which metronidazole is the most effective, are recommended (5,6). Trichomonas vaginalis is the most common non-viral sexually transmitted disease, affecting more than 200 million people annu-ally around the world (7,8). Given the persistent symptoms of this vaginitis, drugs such as metronidazole, tinidazole, and secnidazole are used to treat it.…”

Purification and Characterization of a Novel Intracellular Sucrase Enzyme ofLeishmania donovaniPromastigotes

“…Interestingly very recently Dirkx et al [ 24 ] reported that the flagellated protozoan Trichomonas vaginalis genome contains nearly 11 putative sucrose transporters and a putative ß-fructofuranosidase (invertase). Thus, the machinery for both uptake and cleavage of intracellular sucrose appears to be present in the protozoa as the cell lysates retain invertase activity.…”