Study of the physiological characterization of three lactic acid bacteria and their isogenic ldh deletion mutants
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Lactic acid bacteria generally use homolactic fermentation for generation of ATP. Here we studied the role of the lactate dehydrogenase enzyme on the general physiology of the three lactic acid bacteria Lactococcus lactis, Enterococcus faecalis and Streptococcus pyogenes. Surprisingly deletion of the ldh genes hardly affected the growth rate in chemically defined medium, however growth rate was affected in rich medium. Furthermore, deletion of ldh affected the ability for utilization of various substrates as a carbon source. A switch to mixed acid fermentation was observed in glucose-limited continuous growth and was dependent on growth rate for S. pyogenes and pH dependeny in E. faecalis. In S. pyogenes and L. lactis a change in pH resulted in a clear change in Yatp demand. The pH that showed the highest Yatp corresponded to the pH of the natural habitat of the organisms.
SEEK ID: https://fairdomhub.org/studies/55
Investigation of glycolysis and pyruvate branching of three lactic acid bacteria
Projects: SysMO-LAB
Study position:
Experimentalists: Martijn Bekker
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Created: 11th Jun 2010 at 08:56
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Institutions: University of Amsterdam
Expertise: Microbiology, Biochemistry, Molecular Biology, Escherichia coli, HPLC
Tools: Biochemistry, Molecular Biology, Fermentation, Chromatography
Martijn Bekker (1979) was born in Amstelveen (The Netherlands). He started his studies in biology in 1997 at the University of Amsterdam, and graduated in 2003 with specializations in molecular microbiology and in immunology. The internships during his undergraduate studies were carried out in the labs of Prof. dr. B. Oudega (VU, Amsterdam, The Netherlands) and Prof. dr. F. Heffron (OHSU, Portland, Oregon, USA). He continued with his graduate studies in 2003 in the Laboratory for Molecular Microbial ...
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
S. pyogenes M49 (591), E. faecalis V583, and L. lacis NZ9000 and their isogenic ldh deletion mutants were grown glucose free CDM-LAB medium in BIOLOG phenotype microarray plates PM01 and PM02. With this assay the abilitiy of the strains to grow on 190 different carbon sources was determined in 96 well format.
Submitter: Tomas Fiedler
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Investigation of glycolysis and pyruvate branch...
L. lactis, E. faecalis and S. pyogenes are referred to as LAB because of the fact that in the presence of glucose lactate is produced as the main fermentation product . This metabolic pathway is relatively inefficient, since only 2 ATP are generated from one glucose molecule. All three LAB possess the genetic make up for mixed acid fermentation, a more effective way of fermentation generating 3 ATP per molecule of glucose. All three genomes reveal (at least) two genes encoding a lactate dehydrogenase ...
Submitter: Martijn Bekker
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Investigation of glycolysis and pyruvate branch...
Organisms: Lactococcus lactis, Streptococcus pyogenes : M49 (591) (wild-type / wild-type), Enterococcus faecalis : V583 (wild-type / wild-type)
SOPs: No SOPs
Data files: Maximal specific growth rates of the three lact...
Snapshots: No snapshots
L. lactis, S. pyogenes and E. faecalis were grown in C-limited chemostat cultures at various pH's and dilution rates. General flux distribution, yields and other physiological factors were studied.
Submitter: Martijn Bekker
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Investigation of glycolysis and pyruvate branch...
Organisms: Lactococcus lactis, Streptococcus pyogenes : M49 (591) (wild-type / wild-type), Enterococcus faecalis : V583 (wild-type / wild-type)
SOPs: No SOPs
Data files: Characterization of flux distribution and gener..., Characterization of flux distribution and gener..., Characterization of flux distribution and gener...
Snapshots: No snapshots
. L. lactis (NZ9000), E. faecalis (V538) and S. pyogenes (M49) wild type strain and their ldh- mutants were grown in batch cultures at 37°C in anaerobic 96 wells plates in either TH-broth supplemented with 0.5% (w/v) yeast (THY) or a chemically defined medium for LAB (pH 7.4) (CDM-LAB (10)). Both media were buffered with either 100 mM MES buffer or 100 mM MOPS buffer for growth at pH 6.5 and 7.5 respectively.
Creators: Martijn Bekker, Tomas Fiedler
Submitter: Martijn Bekker
S. pyogenes was grown in C-limited cultures at pH 6.7 and 7.5 and at a growth rate of 0.05 and 0.15
Creators: Martijn Bekker, Tomas Fiedler
Submitter: Martijn Bekker
E. faecalis was grown in C-limited cultures at pH 6.7 and 7.5 and at a growth rate of 0.05 and 0.15
Creators: Ibrahim Mehmeti, Maria Jonsson
Submitter: Martijn Bekker
L. lactis was grown in C-limited cultures at pH 6.7 and 7.5 and at a growth rate of 0.05, 0.15 and 0.40
Creator: Martijn Bekker
Submitter: Martijn Bekker
Abstract (Expand)
Authors: , , , , Anja Pritzschke, Nikolai Siemens, , ,
Date Published: 25th Nov 2010
Publication Type: Not specified
PubMed ID: 21097579
Citation: