Quantitative metabolomics analysis of amino acid metabolism in recombinant Pichia pastoris under different oxygen availability conditions
Carnicer Heras, Marc (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Ten Pierick, Angela (Delft University of Technology. Department of Biotechnology (Delft, Països Baixos))
van Dam, Jan (Delft University of Technology. Department of Biotechnology (Delft, Països Baixos))
Heijnen, Joseph J. (Delft University of Technology. Department of Biotechnology (Delft, Països Baixos))
Albiol i Sala, Joan (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
van Gulik, Walter (Delft University of Technology. Department of Biotechnology (Delft, Països Baixos))
Ferrer, Pau (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)

Date:	2012
Abstract:	Background: Environmental and intrinsic stress factors can result in the global alteration of yeast physiology, as evidenced by several transcriptional studies. Hypoxia has been shown to have a beneficial effect on the expression of recombinant proteins in Pichia pastoris growing on glucose. Furthermore, transcriptional profiling analyses revealed that oxygen availability was strongly affecting ergosterol biosynthesis, central carbon metabolism and stress responses, in particular the unfolded protein response. To contribute to the better understanding of the effect and interplay of oxygen availability and foreign protein secretion on central metabolism, a first quantitative metabolomic analysis of free amino acids pools in a recombinant P. pastoris strain growing under different oxygen availability conditions has been performed. Results: The values obtained indicate significant variations in the intracellular amino acid pools due to different oxygen availability conditions, showing an overall increase of their size under oxygen limitation. Notably, even while foreign protein productivities were relatively low (about 40-80 μg Fab/gDCW·h), recombinant protein production was found to have a limited but significant impact on the intracellular amino acid pools, which were generally decreased in the producing strain compared with the reference strain. However, observed changes in individual amino acids pools were not correlated with their corresponding relative abundance in the recombinant protein sequence, but to the overall cell protein amino acid compositional variations. Conclusions: Overall, the results obtained, combined with previous transcriptomic and proteomic analyses provide a systematic metabolic fingerprint of the oxygen availability impact on recombinant protein production in P. pastoris.
Rights:	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.
Language:	Anglès
Document:	Article ; Versió publicada
Subject:	Pichia pastoris ; Metabolite quantification ; Recombinant protein production ; Hypoxia ; Amino acids ; Metabolic burden
Published in:	Microbial cell factories, Vol. 11, N. 83 (June 2012) , p. 1-10, ISSN 1475-2859

DOI: 10.1186/1475-2859-11-83
PMID: 22704468

10 p, 638.7 KB

Additional file 1 1 p, 21.1 KB

Additional file 2 1 p, 264.7 KB

Additional file 3 1 p, 379.8 KB

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