English   español  
Por favor, use este identificador para citar o enlazar a este item: http://hdl.handle.net/10261/128070
Título

Heterologous expression of a fungal sterol esterase/lipase in different hosts: Effect on solubility, glycosylation and production

AutorVaquero, María Eugenia ; Barriuso, Jorge ; Medrano, Francisco Javier; Prieto, Alicia ; Martínez, María Jesús
Palabras claveEscherichia coli
Saccharomyces cerevisiae
Pichia pastoris
Lipase
Sterol esterase
Aggregation behavior
Glycosylation
Fecha de publicacióndic-2015
EditorElsevier
CitaciónJournal of Bioscience and Bioengineering 120 ( 6) 637–643 (2015)
ResumenOphiostoma piceae secretes a versatile sterol-esterase (OPE) that shows high efficiency in both hydrolysis and synthesis of triglycerides and sterol esters. This enzyme produces aggregates in aqueous solutions, but the recombinant protein, expressed in Komagataella (synonym Pichia) pastoris, showed higher catalytic efficiency because of its higher solubility. This fact owes to a modification in the N-terminal sequence of the protein expressed in Pichia pastoris, which incorporated 4–8 additional amino acids, affecting its aggregation behavior. In this study we present a newly engineered P. pastoris strain with improved protein production. We also produced the recombinant protein in the yeast Saccharomyces cerevisiae and in the prokaryotic host Escherichia coli, corroborating that the presence of these N-terminal extra amino acids affected the protein's solubility. The OPE produced in the new P. pastoris strain presented the same physicochemical properties than the old one. An inactive form of the enzyme was produced by the bacterium, but the recombinant esterase from both yeasts was active even after its enzymatic deglycosylation, suggesting that the presence of N-linked carbohydrates in the mature protein is not essential for enzyme activity. Although the yield in S. cerevisiae was lower than that obtained in P. pastoris, this work demonstrates the importance of the choice of the heterologous host for successful production of soluble and active recombinant protein. In addition, S. cerevisiae constitutes a good engineering platform for improving the properties of this biocatalyst.
Descripción29 p.-5 fig.
Versión del editorhttp://dx.doi.org/ 10.1016/j.jbiosc.2015.04.005
URIhttp://hdl.handle.net/10261/128070
DOI10.1016/j.jbiosc.2015.04.005
ISSN1389-1723
E-ISSN1347-4421
Aparece en las colecciones: (CIB) Artículos
Ficheros en este ítem:
Fichero Descripción Tamaño Formato  
JBB-Postprint-Vaquero et al 2015.docPostprint6,19 MBMicrosoft WordVisualizar/Abrir
Mostrar el registro completo
 

Artículos relacionados:


NOTA: Los ítems de Digital.CSIC están protegidos por copyright, con todos los derechos reservados, a menos que se indique lo contrario.