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General Information
Manuscript title Dynamic simulation of an in vitro multi-enzyme system.
PubMed ID 17239859
Journal FEBS Letters
Year 2007
Authors Nobuyoshi Ishii, Yoshihiro Suga, Akiko Hagiya, Hisami Watanabe, Hirotada Mori, Masataka Yoshino, Masaru Tomita
Affiliations Institute for Advanced Biosciences, Keio University, Japan
Keywords dynamic simulation, enzyme kinetics, metabolism, glycolysis, in vitro
Full text article Ishii_2007.pdf
Project name not specified


Experiment Description
Organism Escherichia coli
Strain K-12 AG1
Data type time-series data of metabolites
Data units mM
Execution date not specified


Experimental Details
Temperature (°C) 37.0
pH not specified
Carbon source glucose,
Culture mode batch
Process condition aerobic
Dilution rate (h⁻¹)
Working volume (L) 0.2
Biomass concentration (g/L) not specified
Medium composition

LB medium supplemented with 50 µg/ml chloramphenicol to OD600=0.5

General protocol information Sampling method: Samples were obtained 2 h after addition of 0.1 mM isopropyl-b-D-thiogalactopyranoside (IPTG)

Quenching procedure: cells were collected by centrifugation (6000 r.p.m., 5 min at 4 ºC) and resuspended in 10 ml of cold buffer I [50 mM sodium phosphate (pH 7.0), 200 mM NaCl, Protease inhibitor (Complete Mini EDTA-Free, Roche Diagnostics, Basel, Switzerland)].

Extraction technique: sonication

Sample analyzing method: enzymatic

Methods description - Notes

Measurement of metabolite concentrations - A sample was added to 100 mM MOPS buffer (pH 7.2) containing 0.1 mM NADH, 20 mM MgCl2, 5 mM ATP, and 2 mM DTT (I), and maintained at 37 ºC. Then, 1.8 U/ml ALD, 31 U/ml G3PDH, and 90 U/ml TIM (II), 3 U/ml PfkA (III), 18 U/ml Pgi (IV) ...

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Submission and curation

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: 2013-05-13 14:45:37 UTC

Updated: 2020-04-24 16:10:31 UTC

Version: 0

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

Views: 6

Downloads: 1




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