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Metabolic Flux Analysis pp 291–313Cite as

13C-Based Metabolic Flux Analysis of Recombinant Pichia pastoris

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1191))

Abstract

Overexpression of a foreign protein may negatively affect several cell growth parameters, as well as cause cellular stress. Central (or core) metabolism plays a crucial role since it supplies energy, reduction equivalents, and precursor molecules for the recombinant product, cell’s maintenance, and growth needs. However, the number of quantitative physiology studies of the impact of recombinant protein production on the central metabolic pathways of yeast cell factories has been traditionally rather limited, thereby hampering the application of rational strain engineering strategies targeting central metabolism.

The development and application of quantitative physiology and modelling tools and methodologies is allowing for a systems-level understanding of the effect of bioprocess parameters such as growth rate, temperature, oxygen availability, and substrate(s) choice on metabolism, and its subsequent interactions with recombinant protein synthesis, folding, and secretion.

Here, we review the recent developments and applications of 13C-based metabolic flux analysis (13C-MFA) of Pichia pastoris and the gained understanding of the metabolic behavior of this yeast in recombinant protein production bioprocesses. We also discuss the potential of multilevel studies integrating 13C-MFA with other omics analyses, as well as future perspectives on the metabolic modelling approaches to study and design metabolic engineering strategies for improved protein production.

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Acknowledgement

We appreciate the support by the Spanish Ministry of Science and Innovation (CICYT project CTQ2010-15131) and the Catalan Government (contract grant 2009-SGR-281, Xarxa de Referència en Biotecnologia and AGAUR Project grant 2010 CONE3-00063).

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Authors and Affiliations

  1. Escola d’Enginyeria, Edifici Q, Universitat Autònoma de Barcelona, Campus de Bellaterra, 08193, Bellaterra (Cerdanyola del Vallès), Catalonia, Spain

    Pau Ferrer

  2. Department of Chemical Engineering, Escola d’Enginyeria, Universitat Autònoma de Barcelona, Bellaterra (Cerdanyola del Vallès), Catalonia, Spain

    Pau Ferrer & Joan Albiol

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  1. Pau Ferrer
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  2. Joan Albiol
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Correspondence to Pau Ferrer .

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Editors and Affiliations

  1. Centre for Microbial Electrosynthesis (CEMES), Advanced Water Management Centre, University of Queensland, St. Lucia, Brisbane, Queensland, Australia

    Jens O. Krömer

  2. AIBN, University of Queensland, St. Lucia, Brisbane, Queensland, Australia

    Lars K. Nielsen

  3. Biology Department, RWTH Aachen University, Aachen, Germany

    Lars M. Blank

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Ferrer, P., Albiol, J. (2014). 13C-Based Metabolic Flux Analysis of Recombinant Pichia pastoris . In: Krömer, J., Nielsen, L., Blank, L. (eds) Metabolic Flux Analysis. Methods in Molecular Biology, vol 1191. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1170-7_17

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  • DOI: https://doi.org/10.1007/978-1-4939-1170-7_17

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