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Manuscript title Glucose-methanol co-utilization in Pichia pastoris studied by metabolomics ans instationary 13C flux analysis.
PubMed ID 23448228
Journal BMC Systems Biology
Year 2013
Authors Joel Jordà, Camilo Suarez, Marc Carnicer, Angela ten Pierick, Joseph J Heijnen, Walter van Gulik, Pau Ferrer, Joan Albiol and Aljoscha Wahl
Affiliations Department of Chemical Engineering, Escola d’Enginyeria, Universitat Autònoma de Barcelona, Bellaterra, Cerdanyola del Vallès, Spain
Keywords Pichia pastoris, instationary 13C-metabolic flux analysis, GC-MS, LC-MS
Full text article Downloadarticle Jorda_2013.pdf
Project name not specified

Experiment Description info

Organism Pichia pastoris
Strain X-33
Data type flux measurements
Data units (mmol/gDW.h)
Execution date not specified

Experimental Details info

Temperature (0C) 25.0
pH 5.0
Carbon source glucose,
Culture mode chemostat
Process condition aerobic
Dilution rate (h-1) 0.09
Working volume (L) 1.0
Biomass concentration (g/L) 4.0
Medium composition

7.04 g/l Glucose monohydrate, 1.6 g/l Methanol, 0.915 g/l citric acid, 2 g/l (NH4)2 HPO4, 0.3 g/l MgSO4.7H2O, 1.4 g/l KH2PO4, 0.01 g/l CaCl2 · 2H2O, 0.5 ml/l trace salt solution, 0.3 ml/l biotin (0.2 g/l)

General protocol information Flux analysis method: 13C constrained MFA

Platform: GC-MS, LC-MS

Methods description - Notes

Labelling experiment - After a minimum of five residence times of continuous cultivation, the feed was switched to the labelled medium. The composition of the labelling feed was based on the approach proposed by Nöh and Wiechert [1]. The feed contained for glucose, 80% [1-13C1] and 20% [U-13C6] and for methanol, 100% [U-13C1]. Immediately after switching to labelled medium feed, more than 10 samples were taken during the first five minutes; thereafter, 10 more samples were taken evenly distributed until 6 hours of labelling feed. The labelling experiment was performed in two independent chemostat cultivation replicates.
13C-based metabolic flux analysis (13C-MFA) - The metabolic network used for 13C-based metabolic flux analyses was derived from the metabolic model described in [2] for P. pastoris. This initial model was further extended to include trehalose metabolism and the pyruvate dehydrogenase bypass reactions, as well as reversible reactions for Ala, Asp and Glu synthesis.

---------References----------
[1] Nöh K, Wiechert W: Experimental design principles for isotopically instationary 13C labeling experiments. Biotechnol Bioeng 2006, 94:234–251. http://doi.org/bxmkn8
[2] Jordà J, Jouhten P, Cámara E, Maaheimo H, Albiol J, Ferrer P: Metabolic flux profiling of recombinant protein secreting Pichia pastoris growing on glucose:methanol mixtures. Microb Cell Fact 2012, 11:57. http://doi.org/s6q

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Downloadfluxes KIMODATAID57_v0.xlsx
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Submission and curation info

Entered by Administrator KiMoSysFirst name: Administrator
Affiliation: INESC-ID/IST
Homepage: http://kdbio.inesc-id.pt/kimosys
Interests: mathematical modeling, accessible data, use of data

Created 2013-06-04 11:32:58 UTC

Updated 2014-06-12 22:33:36 UTC

Version 0

Status (reviewed) 2013-12-06 17:17:38 UTC




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