Chapter title |
(13)C-Based Metabolic Flux Analysis of Recombinant Pichia pastoris.
|
---|---|
Chapter number | 17 |
Book title |
Metabolic Flux Analysis
|
Published in |
Methods in molecular biology, January 2014
|
DOI | 10.1007/978-1-4939-1170-7_17 |
Pubmed ID | |
Book ISBNs |
978-1-4939-1169-1, 978-1-4939-1170-7
|
Authors |
Pau Ferrer, Joan Albiol |
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 (13)C-based metabolic flux analysis ((13)C-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 (13)C-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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