Heterologous Expression of LDHs from different lactic acid bacteria in Escherichia coli DH5α.
Assessment of kinetic parameters of LDH to include in a catabolic model.
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Created: 2nd Apr 2012 at 14:34
Last updated: 16th May 2014 at 10:13
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Version 2 (latest) Created 16th May 2014 at 10:05 by Silvio Hering
biosample updated.
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Projects: SysMO-LAB
Institutions: University of Rostock
Postdoc @ Institute of Medical Microbiology, University of Rostock
Projects: SysMO-LAB
Institutions: University of Rostock
Expertise: Molecular microbiology
Tools: Biochemistry and protein analysis, Genetic modification, Metabolomics
SysMO is a European transnational funding and research initiative on "Systems Biology of Microorganisms".
The goal pursued by SysMO was to record and describe the dynamic molecular processes going on in unicellular microorganisms in a comprehensive way and to present these processes in the form of computerized mathematical models.
Systems biology will raise biomedical and biotechnological research to a new quality level and contribute markedly to progress in understanding. Pooling European research ...
Projects: BaCell-SysMO, COSMIC, SUMO, KOSMOBAC, SysMO-LAB, PSYSMO, SCaRAB, MOSES, TRANSLUCENT, STREAM, SulfoSys, SysMO DB, SysMO Funders, SilicoTryp, Noisy-Strep
Web page: http://sysmo.net/
Comparative Systems Biology: Lactic Acid Bacteria
Programme: SysMO
Public web page: http://www.sysmo.net/index.php?index=57
Challenge: Comparative analyses, as demonstrated by comparative genomics and bioinformatics, are extremely powerful for (i) transfer of information from (experimentally) well-studied organisms to the other organisms, and (ii) when coupled to functional and phenotypic information, insight in the relative importance of components to the observed differences and simalities. The central principle of this proposal is that important aspects of the functional differences between organisms derive not ...
Submitter: Martijn Bekker
Studies: Comparative modeling and phosphate dependence flux distributions and glu..., Kinetics of L-lactate dehydrogenase from S. pyogenes, E. faecalis and L...., Reconstructing the metabolic pathways of S. pyogenes and E. faecalis fro..., Study of the physiological characterization of three lactic acid bacteri...
Assays: BIOLOG substrate utilization assay, Genome-Scale Model Enterococcus faecalis V583, Genome-scale model of Streptococcus pyogenes, Global sensitivity analysis, Glucose pulsed L. lactis, Glucose pulsed S. pyogenes, Kinetics of L-lactate dehydrogenase from L. lactis, Kinetics of L-lactate dehydrogenase from S. pyogenes, E. faecalis, and L..., Maximal specific growth rates of the three lactic acid bacteria and thei..., Model of L. lactis glycolysis, Physiological characterization of Lactic acid bacteria grown in C-limite..., Regulation of the activity of lactate dehydrogenases from four lactic ac...
Snapshots: No snapshots
The two lactic acid bacteria L. lactis and S. pyogenes were studied with respect to the concentration of intracellular metabolites involved in glycolysis in time upon a glucose pulse. Models that describe this behavior are also constructed
Submitter: Martijn Bekker
Investigation: Investigation of glycolysis and pyruvate branch...
Assays: Global sensitivity analysis, Glucose pulsed L. lactis, Glucose pulsed S. pyogenes, Kinetics of L-lactate dehydrogenase from L. lactis, Model of L. lactis glycolysis, Regulation of the activity of lactate dehydrogenases from four lactic ac...
Snapshots: No snapshots
The Lactate dehydrogenases (LDH) are key metabolic enzymes in lactic acid bacteria (LAB). The LDH ( E.C. 1.1.1.27) catalyzes the reaction of pyruvate and NADH into lactate and NAD+.We have carried out an experimental and computational study of the effects of fructose-1,6-bisphosphate (FBP), phosphate (Pi) and ionic strength (NaCl concentration) on 3 LDHs from 3 LABs studied at pH 6 and pH 7.
Submitter: Silvio Hering
Investigation: Investigation of glycolysis and pyruvate branch...
Assays: Kinetics of L-lactate dehydrogenase from S. pyogenes, E. faecalis, and L...
Snapshots: No snapshots
Measurements on Km, Vmax and allosteric activation or inhibition of the heterologously expressed (E. coli) and purifiied main L-lactate dehydrogenase
Submitter: Martijn Bekker
Assay type: Enzymatic Assay
Technology type: Enzymatic Activity Measurements
Investigation: Investigation of glycolysis and pyruvate branch...
Organisms: Lactobacillus plantarum : WCFS1 (wild-type / wild-type), Enterococcus faecalis : V583 (wild-type / wild-type), Streptococcus pyogenes : M49 (591) (wild-type / wild-type)
SOPs: Measurement of LDH activity
Data files: The effects of fructose-1,6-bisphosphate, phosp...
Snapshots: No snapshots
Despite high similarity in sequence and catalytic properties, the L-lactate dehydrogenases (LDH) in lactic acid bacteria (LAB) display differences in their regulation which may arise from their adaptation to different habitats. We combined experimental and computational approaches to investigate the effects of fructose-1,6-bisphosphate (FBP), phosphate (Pi) and ionic strength (NaCl concentration) on 6 LDHs from 4 LABs studied at pH 6 and pH 7. We find: (1) The extent of activation by FBP (Kact) ...
Submitter: Anna Feldman-Salit
Biological problem addressed: Metabolism
Investigation: Investigation of glycolysis and pyruvate branch...
Organisms: Lactococcus lactis, Streptococcus pyogenes, Enterococcus faecalis, Lactobacillus plantarum
Models: Part 1: Comparative modeling of 3D structures o..., Part 2: Computation of electrostatic potentials..., Part 3: Comparison of electrostatic potentials ..., Part 4: Activation / Inhibition Effect on LDHs ..., Part 5: Algorithm to computationally estimate t...
SOPs: No SOPs
Data files: 3D-sturctures of Lactate Dehydrogenase from 4 L..., Activation / Inhibition of 4 LDH enzymes in the..., Comparison of electrostatic potentials of LDH e..., Electrostatic potentials of four LDH enzymes fr..., Example for the binding energy computation of p..., Measurements on kinetics of L-LDHs from differe..., The effects of fructose-1,6-bisphosphate, phosp...
Snapshots: No snapshots