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General Information
Manuscript title Development of quantitative metabolomics for Pichia pastoris.
PubMed ID 22448155
Journal Metabolomics
Year 2012
Authors Marc Carnicer, Andre B. Canelas, Angela ten Pierick, Zhen Zeng, Jan van Dam, Joan Albiol, Pau Ferrer, Joseph J. Heijnen, Walter van Gulik
Affiliations Department of Chemical Engineering, Universitat Autonoma de Barcelona, 08193 Bellaterra (Cerdanyola del Valle`s), Spain
Keywords Pichia pastoris, Metabolite quantification, Quenching, Chemostat
Full text article Carnicer_2012.pdf
Project name not specified

Experiment Description
Organism Pichia pastoris
Strain X-33
Data type metabolites at steady-state
Data units µmol/gDCW
Execution date not specified

Experimental Details
Temperature (°C) 25
pH 5.0
Carbon source glucose
Culture mode chemostat
Process condition aerobic
Dilution rate (h⁻¹) 0.1
Working volume (L) 4.0
Biomass concentration (g/L) ≈ 4.5 g/l
Medium composition

Derived from previously described media [1]. The composition of the batch medium was: 8 g/l glycerol, 0.9 g/l citric acid monohydrate, 12.6 g/l (NH4)2HPO4, 0.5 g/l MgSO4 7H20, 1.5 g/l KH2PO4, 0.02 g/l CaCl2 2H20, 5 ml/l trace salt solution, 2 ml/l Biotin solution (0.2 g/l). The composition of the chemostat medium was: 8.80 g/l glucose monohydrate, 0.92 g/l citric acid monohydrate, 2 g/l (NH4)2HPO4, 0.3 g/l MgSO4 7H20, 1.4 g/l KH2PO4, 0.01 g/l CaCl2 2H20, 0.5 ml/l trace salt solution, 0.3 ml/l Biotin (0.2 g/l). The trace salts solution was the same as described previously [1].

General protocol information Sampling method: samples for intracellular metabolite analysis were taken using a dedicated rapid-sampling setup [2].

Quenching procedure: approximately, 0.63 ± 0.01 g of broth was rapidly withdrawn and immediately injected in 5 ml of precooled quenching solution.

Extraction technique: hot ethanol

Sample analyzing method: GC-MS, LC-ESI-MS

Methods description - Notes

Sampling: The tubes were quickly mixed by vortexing and introduced in the filtration unit after weighting the tube [3]. All sampling tubes were weighted before and after the sampling procedure in order to determine the exact amount of sample taken. Briefly, the cell suspensi ...

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Entered by: Administrator KiMoSysFirst name: Administrator
Affiliation: INESC-ID/IST
Interests: mathematical modeling, accessible data, use of data

Created: 2018-08-01 15:36:45 UTC

Updated: 2020-04-24 16:10:36 UTC

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Status: (reviewed) 2018-08-01 15:37:19 UTC

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