Transcriptome-Based Identification of a Functional Fasciola hepatica Carboxylesterase B


Por: Pedroza-Gomez, Yaretzi J., Cossio-Bayugar, Raquel, Aguilar-Diaz, Hugo, Scarcella, Silvana, Reynaud, Enrique, Sanchez-Carbente, Maria del Rayo, Narvaez-Padilla, Veronica, Miranda-Miranda, Estefan

Publicada: 1 nov 2021
Resumen:
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.

Filiaciones:
Pedroza-Gomez, Yaretzi J.:
 Centro Nacional de Investigaciones Disciplinarias en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales Agrícolas y Pecuarias (INIFAP), Boulevard Cuauhnahuac No. 8534, Jiutepec, Morelos, 62550, Mexico

Cossio-Bayugar, Raquel:
 Centro Nacional de Investigaciones Disciplinarias en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales Agrícolas y Pecuarias (INIFAP), Boulevard Cuauhnahuac No. 8534, Jiutepec, Morelos, 62550, Mexico

Aguilar-Diaz, Hugo:
 Centro Nacional de Investigaciones Disciplinarias en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales Agrícolas y Pecuarias (INIFAP), Boulevard Cuauhnahuac No. 8534, Jiutepec, Morelos, 62550, Mexico

Scarcella, Silvana:
 Facultad de Ciencias Veterinarias, Universidad Nacional del Centro de la Provincia de Buenos Aires, Campus Universitario, Tandil, 7000, Argentina

Reynaud, Enrique:
 Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Avenida Universidad, 2001, Apartado Postal 510–3, Cuernavaca, Morelos, 62210, Mexico

Sanchez-Carbente, Maria del Rayo:
 Centro de Investigación en Biotecnología de la Universidad Autónoma del Estado de Morelos, Av. Universidad 1001, Cuernavaca, Morelos, 62209, Mexico

Narvaez-Padilla, Veronica:
 Centro de Investigación en Dinámica Celular de la Universidad Autónoma del Estado de Morelos, Cuernavaca Morelos México, Av. Universidad 1001, Chamilpa, Cuernavaca, Morelos, 62210, Mexico

Miranda-Miranda, Estefan:
 Centro Nacional de Investigaciones Disciplinarias en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales Agrícolas y Pecuarias (INIFAP), Boulevard Cuauhnahuac No. 8534, Jiutepec, Morelos, 62550, Mexico
ISSN: 20760817





Pathogens
Editorial
MDPI AG, ST ALBAN-ANLAGE 66, CH-4052 BASEL, SWITZERLAND, Suiza
Tipo de documento: Article
Volumen: 10 Número: 11
Páginas:
WOS Id: 000726226200001
ID de PubMed: 34832612
imagen gold, Green Published, Gold