title: Production of a sterol esterase from Ophiostoma piceae in batch and fed-batch bioprocesses using different Pichia pastoris phenotypes as cell factory.
abstract: The potential biotechnological applications for the Ophiostoma piceae sterol esterase (OPE) are conditioned to the availability of high enzyme amounts at low prices. This enzyme is a versatile biocatalyst with different biotechnological applications. In this work a systematic study on its heterologous production in different Pichia pastoris strains and operational strategies is presented. The best results were obtained using an AOX1 defective yeast strain in a fed-batch bioprocess using methanol as inducer substrate at a set point of 2.5 g L(-1) and sorbitol as cosubstrate by means of a preprogramed exponential feeding rate at a μ = 0.02 h(-1) , reaching 30 U mL(-1) of enzyme and a volumetric productivity of 403.5 U L(-1) h(-1) . These values are twofold higher than those obtained with a Mut(+) phenotype using methanol a sole carbon source. OPE was the main protein secreted by the yeast, 55% for Mut(s) versus 25% for Mut(+.)
authors.affiliation: Environmental Biology Dept., Centro de Investigaciones Biológicas (CIB), Spanish National Research Council (CSIC), Ramiro de Maeztu 9, Madrid, 28040, Spain.
authors.forename: Víctor Barba
authors.initials: VB
authors.lastname: Cedillo
authors.forename: María Jesús
authors.initials: MJ
authors.lastname: Martínez
authors.forename: Carolina
authors.initials: C
authors.lastname: Arnau
authors.forename: Francisco
authors.initials: F
authors.lastname: Valero
mesh_terms: D019149:Bioreactors / Q000382:microbiology*; D001709:Biotechnology; D003470:Culture Media; D005656:Fungal Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D055156:Ophiostoma / Q000201:enzymology* / Q000235:genetics; D010641:Phenotype; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002787:Sterol Esterase / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D003470:Culture Media; D005656:Fungal Proteins; D011994:Recombinant Proteins; D002787:Sterol Esterase
keywords: ascomycete; esterase; lipase; recombinant protein