NAD+ and NADH Kinetics, using 13C NMR

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SEEK ID: https://demo.fairdomhub.org/assays/9

Class: Experimental Assay

Submitter: Katy Wolstencroft

Projects: SysMO DB

Investigation: Glycolitic Flux in Lactococcus lactis

Study: The effects of Redox Charge on Glycolytic Flux

Assay type: Glucose Pulse

Technology type: NMR

Organisms: Escherichia coli

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Katy Wolstencroft

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Views: 3971

Created: 8th Aug 2011 at 13:52

Last updated: 26th Oct 2017 at 09:57

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Katy Wolstencroft

Projects: SulfoSys, SysMO DB

Institutions: University of Amsterdam, School of Computer Science, University of Manchester

Expertise: Biochemistry, Bioinformatics, Data Management

Tools: Data Management, Transcriptomics, Databases, Workflows, Web services, Taverna, Ontologies

I am a Research Fellow at the University of Manchester, working in Bioinformatics and Computer Science. I am also a guest researcher at the Vrije Universiteit.My research interests include scientific workflows, semantic discovery, and applying ontology technologies to biological data

SysMO

SysMO is a European transnational funding and research initiative on "Systems Biology of Microorganisms".

The goal pursued by SysMO is 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

Web page: http://sysmo.net/

SysMO DB

The main objectives of SysMO-DB are to: facilitate the web-based exchange of data between research groups within- and inter- consortia, and to provide an integrated platform for the dissemination of the results of the SysMO projects to the scientific community. We aim to devise a progressive and scalable solution to the data management needs of the SysMO initiative, that:

facilitates and maximises the potential for data exchange between SysMO research groups;
maximises the ‘shelf life’ and ...

Programme: SysMO

Public web page: http://www.sysmo-db.org/

Organisms: Saccharomyces cerevisiae, Lactic Acid Bacteria

Glycolitic Flux in Lactococcus lactis

SysMO DB

All creators

Katy Wolstencroft

An investigation into the control mechanisms of glycolytic flux in Lactococcus lactis.

This investigation represents the data and models published by Hoefnagel et al in JWS Online and in M.H.N. Hoefnagel, J. Hugenholtz and J.L. SnoepMol. Biol. Reports 29, 157-161 (2002)

Submitter: Katy Wolstencroft

Studies: The effects of Redox Charge on Glycolytic Flux

Assays: NAD+ and NADH Kinetics, using 13C NMR, Time dependent responses of glycolytic intermediates in a detailed glyco...

Snapshots: No snapshots

Scales: Not specified

Created: 8th Aug 2011 at 13:47, Last updated: 4th Nov 2011 at 10:06

The effects of Redox Charge on Glycolytic Flux

SysMO DB

All creators

Katy Wolstencroft

A study which combines mathematical modelling and experimental measurements to determine the effects of redox charge on glycolytic flux

Submitter: Katy Wolstencroft

Investigation: Glycolitic Flux in Lactococcus lactis

Assays: NAD+ and NADH Kinetics, using 13C NMR, Time dependent responses of glycolytic intermediates in a detailed glyco...

Snapshots: No snapshots

Scales: Not specified

Created: 8th Aug 2011 at 13:50, Last updated: 4th Nov 2011 at 10:06

Is the glycolytic flux in Lactococcus lactis primarily controlled by the redox charge? Kinetics of NAD(+) and NADH pools determined in vivo by 13C NMR

SysMO DB

Abstract (Expand)

The involvement of nicotinamide adenine nucleotides (NAD(+), NADH) in the regulation of glycolysis in Lactococcus lactis was investigated by using (13)C and (31)P NMR to monitor in vivo the kinetics … of the pools of NAD(+), NADH, ATP, inorganic phosphate (P(i)), glycolytic intermediates, and end products derived from a pulse of glucose. Nicotinic acid specifically labeled on carbon 5 was synthesized and used in the growth medium as a precursor of pyridine nucleotides to allow for in vivo detection of (13)C-labeled NAD(+) and NADH. The capacity of L. lactis MG1363 to regenerate NAD(+) was manipulated either by turning on NADH oxidase activity or by knocking out the gene encoding lactate dehydrogenase (LDH). An LDH(-) deficient strain was constructed by double crossover. Upon supply of glucose, NAD(+) was constant and maximal (approximately 5 mm) in the parent strain (MG1363) but decreased abruptly in the LDH(-) strain both under aerobic and anaerobic conditions. NADH in MG1363 was always below the detection limit as long as glucose was available. The rate of glucose consumption under anaerobic conditions was 7-fold lower in the LDH(-) strain and NADH reached high levels (2.5 mm), reflecting severe limitation in regenerating NAD(+). However, under aerobic conditions the glycolytic flux was nearly as high as in MG1363 despite the accumulation of NADH up to 1.5 mm. Glyceraldehyde-3-phosphate dehydrogenase was able to support a high flux even in the presence of NADH concentrations much higher than those of the parent strain. We interpret the data as showing that the glycolytic flux in wild type L. lactis is not primarily controlled at the level of glyceraldehyde-3-phosphate dehydrogenase by NADH. The ATP/ADP/P(i) content could play an important role.

Authors: Ana Rute Neves, Rita Ventura, Nahla Mansour, Claire Shearman, Michael J Gasson, Christopher Maycock, Ana Ramos, Helena Santos

Date Published: 13th May 2002

Publication Type: Not specified

PubMed ID: 12011086

Citation:

Scales: Not specified

Created: 8th Aug 2011 at 13:53, Last updated: 8th Aug 2011 at 13:53