Macromolecular and elemental composition analysis and extracellular metabolite balances of Pichia pastoris growing at different oxygen levels

Carnicer Heras, Marc; Baumann, Kristin; Töplitz, Isabelle; Sánchez-Ferrando, Francisco; Mattanovich, Diethard; Ferrer, Pau; Albiol i Sala, Joan

doi:10.1186/1475-2859-8-65

Cita bibliogràfica -- Enllaç permanent: https://ddd.uab.cat/record/112885

Web of Science: 96 cites, Scopus: 96 cites, Google Scholar: cites

Macromolecular and elemental composition analysis and extracellular metabolite balances of Pichia pastoris growing at different oxygen levels
Carnicer Heras, Marc

(Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Baumann, Kristin (Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Töplitz, Isabelle (University of Applied Sciences. School of Bioengineering (Viena, Àustria))
Sánchez-Ferrando, Francisco (Universitat Autònoma de Barcelona. Departament de Química)
Mattanovich, Diethard

(University of Natural Resources and Life Sciences. Department of Biotechnology (Viena, Àustria))
Ferrer, Pau

(Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)
Albiol i Sala, Joan

(Universitat Autònoma de Barcelona. Departament d'Enginyeria Química, Biològica i Ambiental)

Data:	2009
Resum:	Background: Analysis of the cell operation at the metabolic level requires collecting data of different types and to determine their confidence level. In addition, the acquired information has to be combined in order to obtain a consistent operational view. In the case of Pichia pastoris, information of its biomass composition at macromolecular and elemental level is scarce particularly when different environmental conditions, such as oxygen availability or, genetic backgrounds (e. g. recombinant protein production vs. non production conditions) are compared. Results: P. pastoris cells growing in carbon-limited chemostat cultures under different oxygenation conditions (% O2 in the bioreactor inlet gas: 21%, 11% and 8%, corresponding to normoxic, oxygen-limiting and hypoxic conditions, respectively), as well as under recombinant protein (antibody fragment, Fab) producing and non-producing conditions, were analyzed from different points of view. On the one hand, the macromolecular and elemental composition of the biomass was measured using different techniques at the different experimental conditions and proper reconciliation techniques were applied for gross error detection of the measured substrates and products conversion rates. On the other hand, fermentation data was analyzed applying elemental mass balances. This allowed detecting a previously missed by-product secreted under hypoxic conditions, identified as arabinitol (aka. arabitol). After identification of this C5 sugar alcohol as a fermentation by-product, the mass balances of the fermentation experiments were validated. Conclusions: After application of a range of analytical and statistical techniques, a consistent view of growth parameters and compositional data of P. pastoris cells growing under different oxygenation conditions was obtained. The obtained data provides a first view of the effects of oxygen limitation on the physiology of this microorganism, while recombinant Fab production seems to have little or no impact at this level of analysis. Furthermore, the results will be highly useful in other complementary quantitative studies of P. pastoris physiology, such as metabolic flux analysis.
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial i la comunicació pública de l'obra, sempre que no sigui amb finalitats comercials, i sempre que es reconegui l'autoria de l'obra original. No es permet la creació d'obres derivades.
Llengua:	Anglès
Document:	Article ; Versió publicada
Publicat a:	Microbial cell factories, Vol. 8, N. 65 (December 2009) , p. 1-14, ISSN 1475-2859