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Production of a sterol esterase from Ophiostoma piceae in batch and fed‐batch bioprocesses using different Pichia pastoris phenotypes as cell factory
Author(s) -
Cedillo Víctor Barba,
Martínez María Jesús,
Arnau Carolina,
Valero Francisco
Publication year - 2014
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.1939
Subject(s) - pichia pastoris , bioprocess , yeast , biochemistry , bioprocess engineering , chemistry , heterologous , enzyme , pichia , biology , recombinant dna , microbiology and biotechnology , gene , paleontology
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 +. © 2014 American Institute of Chemical Engineers Biotechnol. Prog ., 30:1012–1020, 2014