Per citar aquest document: http://ddd.uab.cat/record/146866
Quantitative Metabolomics and Instationary 13C-Metabolic Flux Analysis Reveals Impact of Recombinant Protein Production on Trehalose and Energy Metabolism in Pichia pastoris
Jordà Murria, Joel (Universitat Autònoma de Barcelona. Departament d’Enginyeria Química)
Cueto Rojas, Hugo (Delft University of Technology. Department of Biotechnology)
Carnicer Heras, Marc (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química)
Aljoscha Wahl (Delft University of Technology. Department of Biotechnology)
Ferrer i Alegre, Pau (Universitat Autònoma de Barcelona. Departament d’Enginyeria Química)
Albiol i Sala, Joan (Universitat Autonoma de Barcelona. Departament d'Enginyeria Química)

Data: 2014
Resum: Pichia pastoris has been recognized as an effective host for recombinant protein production. In this work, we combine metabolomics and instationary 13C metabolic flux analysis (INST 13C-MFA) using GC-MS and LC-MS/MS to evaluate the potential impact of the production of a Rhizopus oryzae lipase (Rol) on P. pastoris central carbon metabolism. Higher oxygen uptake and CO2 production rates and slightly reduced biomass yield suggest an increased energy demand for the producing strain. This observation is further confirmed by 13C-based metabolic flux analysis. In particular, the flux through the methanol oxidation pathway and the TCA cycle was increased in the Rol-producing strain compared to the reference strain. Next to changes in the flux distribution, significant variations in intracellular metabolite concentrations were observed. Most notably, the pools of trehalose, which is related to cellular stress response, and xylose, which is linked to methanol assimilation, were significantly increased in the recombinant strain.
Drets: Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original. Creative Commons
Llengua: Anglès
Document: article ; recerca ; publishedVersion
Matèria: Pichia pastoris ; Instationary 13C-metabolic flux analysis ; Recombinant protein production ; Trehalose ; Metabolome ; GC-MS ; LC-MS
Publicat a: Metabolites, Vol. 4 (2014) , p. 281-299, ISSN 2218-1989

DOI: 10.3390/metabo4020281


19 p, 492.8 KB

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