The effect of hypoxia on the lipidome of recombinant Pichia pastoris
Adelantado Vallvé, Núria (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Tarazona, Pablo (Georg-August University. Albrecht-von-Haller-Institute for Plant Sciences)
Grillitsch, Karlheinz (Austrian Centre of Industrial Biotechnology (ACIB))
Garcia-Ortega, Xavier (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Monforte, Sergi (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Valero Barranco, Francisco (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Feussner, Ivo (Georg-August University. Albrecht-von-Haller-Institute for Plant Sciences)
Daum, Günther (Graz University of Technology. Institute of Biochemistry)
Ferrer, Pau (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)

Fecha:	2017
Resumen:	Cultivation of recombinant Pichia pastoris (Komagataella sp. ) under hypoxic conditions has a strong positive effect on specific productivity when the glycolytic GAP promoter is used for recombinant protein expression, mainly due to upregulation of glycolytic conditions. In addition, transcriptomic analyses of hypoxic P. pastoris pointed out important regulation of lipid metabolism and unfolded protein response (UPR). Notably, UPR that plays a role in the regulation of lipid metabolism, amino acid metabolism and protein secretion, was found to be upregulated under hypoxia. To improve our understanding of the interplay between lipid metabolism, UPR and protein secretion, the lipidome of a P. pastoris strain producing an antibody fragment was studied under hypoxic conditions. Furthermore, lipid composition analyses were combined with previously available transcriptomic datasets to further understand the impact of hypoxia on lipid metabolism. Chemostat cultures operated under glucose-limiting conditions under normoxic and hypoxic conditions were analyzed in terms of intra/extracellular product distribution and lipid composition. Integrated analysis of lipidome and transcriptome datasets allowed us to demonstrate an important remodeling of the lipid metabolism under limited oxygen availability. Additionally, cells with reduced amounts of ergosterol through fluconazole treatment were also included in the study to observe the impact on protein secretion and its lipid composition. Our results show that cells adjust their membrane composition in response to oxygen limitation mainly by changing their sterol and sphingolipid composition. Although fluconazole treatment results a different lipidome profile than hypoxia, both conditions result in higher recombinant protein secretion levels. The online version of this article (doi:10. 1186/s12934-017-0699-4) contains supplementary material, which is available to authorized users.
Ayudas:	Ministerio de Economía y Competitividad CTQ2013-42391-R Agència de Gestió d'Ajuts Universitaris i de Recerca 2014/SGR-452
Derechos:	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.
Lengua:	Anglès
Documento:	Article ; recerca ; Versió publicada
Materia:	Lipidomics ; Pichia pastoris ; Hypoxia ; Recombinant protein production ; Antibody fragment ; Protein secretion ; Unfolded protein response
Publicado en:	Microbial cell factories, Vol. 16 (May 2017) , art. 86, ISSN 1475-2859

DOI: 10.1186/s12934-017-0699-4
PMID: 28526017

15 p, 2.0 MB

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