DataEntryID 57
    General information
      Manuscript title: Glucose-methanol co-utilization in Pichia pastoris studied by metabolomics ans instationary 13C flux analysis.
      PubMed ID: http://www.ncbi.nlm.nih.gov/pubmed/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: https://kimosys.org/rails/active_storage/blobs/eyJfcmFpbHMiOnsibWVzc2FnZSI6IkJBaHBBbllFIiwiZXhwIjpudWxsLCJwdXIiOiJibG9iX2lkIn19--df9494fe480f61ee916e2314536c9d5722f55c6b/Jorda_2013.pdf
      Project name: not specified

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

    Experimental details
      Temperature (°C): 25.0
      pH: 5.0
      Carbon source: glucose
      Culture mode: chemostat
      Process condition: aerobic
      Dilution rate (h⁻¹): 0.09
      Working volume: 1.0 L
      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: Type analysis list: 13C constrained MFA; Platform list: GC-MS, LC-MS;
      Methods description: 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 
      Data file: http://kimosys.org/repository/57/download?parameter=1141; 
      Alternative formats: no files uploaded

    Submission and curation
      Entered by: Administrator KiMoSys
      Created: 2013-06-04 11:32:58 UTC
      Updated: 2020-04-24 16:10:32 UTC
      Version: 0
      Status: (reviewed) 2013-12-06 17:17:38 UTC
      Views: 218
      Downloads: 60