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Open Access Research article

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

Michael Dirkx1, Michael P Boyer1, Prajakta Pradhan2, Andrew Brittingham2 and Wayne A Wilson1*

Author Affiliations

1 Department of Biochemistry & Nutrition, Des Moines University, Des Moines, IA 50312, USA

2 Department of Microbiology & Immunology, Des Moines University, Des Moines, IA 50312, USA

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BMC Biochemistry 2014, 15:12  doi:10.1186/1471-2091-15-12

Published: 28 June 2014

Abstract

Background

Trichomonas 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.

Results

We 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.

Conclusions

T. 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.

Keywords:
Trichomonas vaginalis; Carbohydrate utilization; Invertase; Purification