Glycerol metabolism of Pichia pastoris (Komagataella spp.) characterised by 13 C-based metabolic flux analysis

5Citations
Citations of this article
32Readers
Mendeley users who have this article in their library.
Get full text

Abstract

Metabolic flux analysis based on 13 C-derived constraints has proved to be a powerful method for quantitative physiological characterisation of one of the most extensively used microbial cell factory platforms, Pichia pastoris (syn. Komagataella spp.). Nonetheless, the reduced number of carbon atoms and the symmetry of the glycerol molecule has hampered the comprehensive determination of metabolic fluxes when used as the labelled C-source. Moreover, metabolic models typically used for 13 C-based flux balance analysis may be incomplete or misrepresent the actual metabolic network. To circumvent these limitations, we reduced the genome-scale metabolic model iMT1026-v3.0 into a core model and used it for the iterative fitting of metabolic fluxes to the measured mass isotope distribution of proteinogenic amino acids obtained after fractional 13 C labelling of cells with [1,3- 13 C]-glycerol. This workflow allows reliable estimates to be obtained for in vivo fluxes in P. pastoris cells growing on glycerol as sole carbon source, as well as revising previous assumptions concerning its metabolic operation, such as alternative metabolic branches, calculation of energetic parameters and proposed specific cofactor utilisation.

Cite

CITATION STYLE

APA

Tomàs-Gamisans, M., Ødum, A. S. R., Workman, M., Ferrer, P., & Albiol, J. (2019). Glycerol metabolism of Pichia pastoris (Komagataella spp.) characterised by 13 C-based metabolic flux analysis. New Biotechnology, 50, 52–59. https://doi.org/10.1016/j.nbt.2019.01.005

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free