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General Information info

Manuscript title Overexpression of isocitrate lyase-glyoxylate bypass influence on metabolism in Aspergillus niger.
PubMed ID 19271267
Journal Metabolic Engineering
Year 2009
Authors S. Meijer, J Otero, R. Olivares, M.R. Andersen, L. Olsson, J. Nielsen
Affiliations Department for Systems Biology, Center for Microbial Biotechnology, Building 223, Technical University of Denmark, Lyngby, Denmark.
Keywords Aspergillus niger, isocitrate lyase over-expression, glyoxylate bypass
Full text article Downloadarticle Meijer_2009.pdf
Project name not specified

Experiment Description info

Organism Aspergillus niger
Strain N402 (W.T.) and icl overexpression
Data type flux measurements
Data units % substrate uptake
Execution date not specified

Experimental Details info

Temperature (0C) 30.0
pH 2.5
Carbon source glucose,
Culture mode batch
Process condition aerobic
Dilution rate (h-1) not specified
Working volume (L) 2.0
Biomass concentration (g/L) 2.0-4.0
Medium composition

Defined medium: 5 g/L of 1-13C glucose, 10 g/L NaNO3, 3 g/L KH2PO4, 2 g/L MgSO4.7H2O, 2 g/L NaCl, 0.2 g/L CaCl2.2H2O, 0.5 ml/L antifoam and 1 ml/L trace element solution 2. The trace element solution 2 contained 14.3 g/L ZnSO4.7H2O, 2.5 g/L CuSO4.5H2O, 0.5 g/L NiCl2.6H2O, 7 g/L MnCl2.2H2O and 13.8 g/L FeSO4.7H2O.

General protocol information Flux analysis method: 13C constrained MFA

Platform: GC-MS

Methods description - Notes

GC–MS analysis - GC–MS analysis was performed on an Agilent (PaloAlto,CA, USA) HP 6890 gas chromatograph coupled to a HP5973 quadrupole mass selective detector in positive electron impact ionization (EI+) operated at an electron energy of 70 eV. The GC was equipped with a 4.0mm i.d. Siltek gooseneck splitless deactivated liner (Restek, Bellefonte, PA, USA), and a Supelco (Bellefonte,PA,USA) SLB-5 MS column, 15m, 0.25mm i.d., 0.25 µm film. Helium of a purity of 99.999% was used as carrier gas at a constant linear gas velocity of 35 cm/s. Transfer line temperature was 280 ºC, quadrupole temperature 150 ºC and MS source 200 ºC. The GC–MS system was controlled from Agilent MSD Chemstation v. D.01.02.16. For both methods (ECF and DMF/DMA) 1 mL sample was injected in the splitless (30s, split 1:20) mode at 200 ºC using a hot needle.The MS scanrange was m/z 40–400 with 2.88 s/scan. To avoid sample carry-over the syringe was cleaned using five times 5mL acetone, then five times5 mL dichloromethane and finally 3 times rinsing with 3 µL sample.
Metabolic network analysis - The fluxes were determined using an in-house program code in Matlab V7.0.4 , which implements the metabolic network analysis framework from [1].

--------------References------------------
[1] Wiechert, W.,2001. C13 metabolic flux analysis. Metab. Eng. 3, 195–206. http://doi.org/dr89ms

Data file
Downloadfluxes KIMODATAID73_v0.xlsx
Alternative format(s)
KIMODATAID73_wt.txt
KIMODATAID73_overexpression.txt
KIMODATAID73_wt_reaction_SBtab.csv
KIMODATAID73_overexpression_reaction_SBtab.csv
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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 2013-06-23 12:31:56 UTC

Updated 2014-06-13 16:43:43 UTC

Version 0

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




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