Abstract
Bioinformatics analysis of the complete transcriptome of Fasciola hepatica, identified a total of ten putative carboxylesterase transcripts, including a 3146 bp mRNA transcript coding a 2205 bp open reading frame that translates into a protein of 735 amino acids, resulting in a predicted protein mass of 83.5 kDa and a putative carboxylesterase B enzyme. The gene coding for this enzyme was found in two reported F. hepatica complete genomes stretching 23,230 bp, containing two exons of 1282 and 1864 bp, respectively, as well as a 20,084 bp intron between the exons. The enzymatic activity was experimentally assayed on F. hepatica protein extracts by SDS-PAGE zymograms using synthetic chromogenic substrates, confirming both the theoretical molecular weight and carboxylesterase enzymatic activity. Further bioinformatics predicted that this enzyme is an integral component of the cellular membrane that should be active as a 167 kDa homodimer complex and polyacrylamide gel electrophoresis (PAGE) zymograms experiments confirmed the analysis. Additional bioinformatics analysis showed that DNA sequences that code for this particular enzyme are highly conserved in other parasitic trematodes, although they are labeled hypothetical proteins.
Highlights
The liver fluke Fasciola hepatica is a worldwide zoonotic parasitic trematode affecting a vast range of animal hosts, including humans
The bioinformatics analysis of transcript 473 did not find a direct biochemical pathway that may demonstrate an enzymatic effect on benzimidazole-derived anthelmintics; this does not mean that the carboxylesterase B does not interact with secondary metabolites, our study suggests that this enzyme may be part of several metabolic pathways that are implicated in the metabolism of drugs and their metabolites
Our study suggests that the F. hepatica carboxylesterase B is an essential enzyme for the metabolism of the liver fluke; our bioinformatics analysis of this gene showed that the enzyme is highly conserved in parasitic flatworms, suggesting a fundamental role in parasitic helminth physiology
Summary
The liver fluke Fasciola hepatica is a worldwide zoonotic parasitic trematode affecting a vast range of animal hosts, including humans. It is the causative agent of fascioliasis, a parasitic disease highly prevalent in the livestock industry and capable of parasitizing millions of people worldwide [1]. Xenobiotic metabolism is responsible for the defense of most organisms against poisonous effects of naturally occurring toxins and the enzymes responsible for this type of metabolism are the focus of scientific scrutiny because they mediate the ever-increasing tolerance and resistance of parasites against antiparasitic agents that plague livestock and humans [7]
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