Heterologous expression, purification and characterization of three novel esterases secreted by the lignocellulolytic fungus Penicillium purpurogenum when grown on sugar beet pulp
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Archivos
Fecha
2017
Profesor/a Guía
Facultad/escuela
Idioma
en
Título de la revista
ISSN de la revista
Título del volumen
Editor
Elsevier Ltd
Nombre de Curso
Licencia CC
Atribución/Reconocimiento-NoComercial-SinDerivados 4.0 Internacional
CC BY-NC-ND 4.0
Deed
Licencia CC
https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
Resumen
The lignocellulolytic fungus, Penicillium purpurogenum, grows on a variety of natural carbon sources, among them sugar beet pulp. Culture supernatants of P. purpurogenum grown on sugar beet pulp were partially purified and the fractions obtained analyzed for esterase activity by zymograms. The bands with activity on methyl umbelliferyl acetate were subjected to mass spectrometry to identify peptides. The peptides obtained were probed against the proteins deduced from the genome sequence of P. purpurogenum. Eight putative esterases thus identified were chosen for future work. Their cDNAs were expressed in Pichia pastoris. The supernatants of the recombinant clones were assayed for esterase activity, and five of the proteins were active against one or more substrates: methyl umbelliferyl acetate, indoxyl acetate, methyl esterified pectin and fluorescein diacetate. Three of those enzymes were purified, further characterized and subjected to a BLAST search. Based on their amino acid sequence and properties, they were identified as follows: RAE1, pectin acetyl esterase (CAZy family CE 12); FAEA, feruloyl esterase (could not be assigned to a CAZy family) and EAN, acetyl esterase (former CAZy family CE 10). © 2017 Elsevier Ltd
Notas
Indexación: Scopus
Palabras clave
Esterases, Heterologous expression, Pectin, Penicillium purpurogenum, Pichia pastoris
Citación
Carbohydrate Research Volume 443-444, Pages 42 - 482017
DOI
10.1016/j.carres.2017.03.014