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

Màrius Tomàs-Gamisans, Anders Sebastian Rosenkrans Ødum, Mhairi Workman, Pau Ferrer, Joan Albiol

Research output: Contribution to journalArticleResearch

5 Citations (Scopus)

Abstract

© 2019 Elsevier B.V. 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.
Original languageEnglish
Pages (from-to)52-59
Number of pages8
JournalNew Biotechnology
Volume50
DOIs
Publication statusPublished - 25 May 2019

Keywords

  • BIODIESEL
  • C-13-based meabolic flux analysis
  • CARBON SOURCE
  • CRUDE GLYCEROL
  • FED-BATCH CULTIVATION
  • GLYCEROL/METHANOL MIXTURES
  • GROWTH-RATE
  • Genome-Scale metabolic model
  • Glycerol
  • MASS ISOTOPOMER DISTRIBUTIONS
  • Pichia pastorisKomagataella spp
  • RECOMBINANT PROTEIN-PRODUCTION
  • SACCHAROMYCES-CEREVISIAE
  • STOICHIOMETRY

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