Did you know ? If you order before Friday 14h we deliver 90PCT of the the time next Tuesday, GENTAUR another in time delivery

Pubmed ID :26059529
Publication Date : //

Determination of the in vivo NAD:NADH ratio in Saccharomyces cerevisiae under anaerobic conditions, using alcohol dehydrogenase as sensor reaction.


With the current quantitative metabolomics techniques, only whole-cell concentrations of NAD and NADH can be quantified. These measurements cannot provide information on the in vivo redox state of the cells, which is determined by the ratio of the free forms only. In this work we quantified free NAD:NADH ratios in yeast under anaerobic conditions, using alcohol dehydrogenase (ADH) and the lumped reaction of glyceraldehyde-3-phosphate dehydrogenase and 3-phosphoglycerate kinase as sensor reactions. We showed that, with an alternative accurate acetaldehyde determination method, based on rapid sampling, instantaneous derivatization with 2,4 diaminophenol hydrazine (DNPH) and quantification with HPLC, the ADH-catalysed oxidation of ethanol to acetaldehyde can be applied as a relatively fast and simple sensor reaction to quantify the free NAD:NADH ratio under anaerobic conditions. We evaluated the applicability of ADH as a sensor reaction in the yeast Saccharomyces cerevisiae, grown in anaerobic glucose-limited chemostats under steady-state and dynamic conditions. The results found in this study showed that the cytosolic redox status (NAD:NADH ratio) of yeast is at least one order of magnitude lower, and is thus much more reduced, under anaerobic conditions compared to aerobic glucose-limited steady-state conditions. The more reduced state of the cytosol under anaerobic conditions has major implications for (central) metabolism. Accurate determination of the free NAD:NADH ratio is therefore of importance for the unravelling of in vivo enzyme kinetics and to judge accurately the thermodynamic reversibility of each redox reaction.

Authors : Bekers K M , Heijnen J J , van Gulik W M ,

Related products :

Catalog number Product name Quantity
30-896 Lactate dehydrogenase C catalyzes the conversion of L-lactate and NAD to pyruvate and NADH in the final step of anaerobic glycolysis. LDHC is testis-specific and belongs to the lactate dehydrogenase f 0.05 mg
SCH-8203-0050 RABBIT ANTI SACCHAROMYCES CEREVISIAE, Product Type Polyclonal Antibody, Specificity SACCHAROMYCES CEREVISIAE, Target Species Yeast, Host Rabbit, Format Purified, Isotypes Polyclonal IgG, Applica 1 ml
8203-0050 RABBIT ANTI SACCHAROMYCES CEREVISIAE, Product Type Polyclonal Antibody, Specificity SACCHAROMYCES CEREVISIAE, Target Species Yeast, Host Rabbit, Format Purified, Isotypes Polyclonal IgG, Applica 1 ml
18-272-196783 Saccharomyces cerevisiae - Rabbit polyclonal to Saccharomyces cerevisiae Polyclonal 0.2 ml
18-272-196784 Saccharomyces cerevisiae - Rabbit polyclonal to Saccharomyces cerevisiae Polyclonal 0.5 ml
21216 Ratio Works™ pH sensor II *Cell_permeable* 5 mg
21215 Ratio Works™ pH sensor I *Cell_permeable* 5 mg
21215 Ratio Works™ pH sensor I *Cell_permeable* 5 mg
21216 Ratio Works™ pH sensor II *Cell_permeable* 5 mg
21217 Ratio Works™ pH sensor III *Cell_permeable* 5 mg
21217 Ratio Works™ pH sensor III *Cell_permeable* 5 mg
EIAAB26818 Bos taurus,Bovine,NADH dehydrogenase [ubiquinone] flavoprotein 2, mitochondrial,NADH dehydrogenase subunit II,NADH-ubiquinone oxidoreductase 24 kDa subunit,NDUFV2
21303 Ratio Works™ redox sensor *Cell_permeable* 25 mg
21303 Ratio Works™ redox sensor *Cell_permeable* 25 mg
EIAAB26811 CI-51kD,Complex I-51kD,Homo sapiens,Human,NADH dehydrogenase [ubiquinone] flavoprotein 1, mitochondrial,NADH dehydrogenase flavoprotein 1,NADH-ubiquinone oxidoreductase 51 kDa subunit,NDUFV1,UQOR1
84604-16-0 Saccharomyces cerevisiae Saccharomyces 1g
EIAAB26813 Bos taurus,Bovine,CI-51kD,Complex I-51kD,NADH dehydrogenase [ubiquinone] flavoprotein 1, mitochondrial,NADH dehydrogenase flavoprotein 1,NADH-ubiquinone oxidoreductase 51 kDa subunit,NDUFV1,UQOR1
E0681h ELISA kit ADH1,ADH1A,Alcohol dehydrogenase 1A,Alcohol dehydrogenase subunit alpha,Homo sapiens,Human 96T
E0681h ELISA ADH1,ADH1A,Alcohol dehydrogenase 1A,Alcohol dehydrogenase subunit alpha,Homo sapiens,Human 96T
U0681h CLIA ADH1,ADH1A,Alcohol dehydrogenase 1A,Alcohol dehydrogenase subunit alpha,Homo sapiens,Human 96T
U0681Rb CLIA ADH1,Alcohol dehydrogenase 1,Alcohol dehydrogenase subunit alpha,Oryctolagus cuniculus,Rabbit 96T
E0681Rb ELISA ADH1,Alcohol dehydrogenase 1,Alcohol dehydrogenase subunit alpha,Oryctolagus cuniculus,Rabbit 96T
E0681Rb ELISA kit ADH1,Alcohol dehydrogenase 1,Alcohol dehydrogenase subunit alpha,Oryctolagus cuniculus,Rabbit 96T
E0681r ELISA Adh1,Adh-1,Alcohol dehydrogenase 1,Alcohol dehydrogenase A subunit,Rat,Rattus norvegicus 96T
E0681m ELISA kit Adh1,Adh-1,ADH-A2,Alcohol dehydrogenase 1,Alcohol dehydrogenase A subunit,Mouse,Mus musculus 96T


 

GENTAUR Belgium BVBA BE0473327336
Voortstraat 49, 1910 Kampenhout BELGIUM
Tel 0032 16 58 90 45

Fax 0032 16 50 90 45
info@gentaur.com | Gentaur | Gentaur





GENTAUR Ltd.
Unicorn House, Station Cl
Hertfordshire, Potters Bar EN6 1TL
Whetstone London N20 9BH
Tel 020 3393 8531 Fax 020 8445 9411
uk@gentaur.com | Gentaur | Gentaur

 

 




GENTAUR France SARL
9, rue Lagrange, 75005 Paris
Tel 01 43 25 01 50

Fax 01 43 25 01 60
RCS Paris B 484 237 888

SIRET 48423788800017
RIB 30004 00187 00010092253 10
BNP PARIBAS PARIS PL MAUBERT BIC BNPAFRPPPRG
IBAN FR76 3000 4001 8700 0100 9225 310
france@gentaur.com | Gentaur | Gentaur

GENTAUR GmbH
Marienbongard 20
52062 Aachen Deutschland
Support Karolina Elandt
Tel: 0035929830070
Fax: (+49) 241 56 00 47 88

Logistic :0241 40 08 90 86
Bankleitzahl 39050000
IBAN lautet DE8839050000107569353
Handelsregister Aachen HR B 16058
Umsatzsteuer-Identifikationsnummer *** DE 815175831
Steuernummer 201/5961/3925
de@gentaur.com | Gentaur | Gentaur

GENTAUR U.S.A
Genprice Inc, Logistics
547, Yurok Circle
San Jose, CA 95123
CA 95123
Tel (408) 780-0908,
Fax (408) 780-0908,
sales@genprice.com

Genprice Inc, Invoices and accounting
6017 Snell Ave, Ste 357
San Jose, CA 95123




GENTAUR Nederland BV
NL850396268B01 KVK nummer 52327027
Kuiper 1
5521 DG Eersel Nederland
Tel:  0208-080893  Fax: 0497-517897
nl@gentaur.com | Gentaur | Gentaur
IBAN: NL04 RABO 0156 9854 62   SWIFT RABONL2U






GENTAUR Spain
tel:0911876558
spain@gentaur.com | Gentaur | Gentaur






ГЕНТАУЪР БЪЛГАРИЯ
ID # 201 358 931 /BULSTAT
София 1000, ул. "Граф Игнатиев" 53 вх. В, ет. 2
Tel 0035924682280 Fax 0035924808322
e-mail: Sofia@gentaur.com | Gentaur | Gentaur
IBAN: BG11FINV91501014771636
BIC: FINVBGSF

GENTAUR Poland Sp. z o.o.


ul. Grunwaldzka 88/A m.2
81-771 Sopot, Poland
TEL Gdansk 058 710 33 44 FAX  058 710 33 48              

poland@gentaur.com | Gentaur | Gentaur

Other countries

Österreich +43720880899

Canada Montreal +15149077481

Ceská republika Praha +420246019719

Danmark +4569918806

Finland Helsset +358942419041

Magyarország Budapest +3619980547

Ireland Dublin+35316526556

Luxembourg+35220880274

Norge Oslo+4721031366

Sverige Stockholm+46852503438

Schweiz Züri+41435006251

US New York+17185132983

GENTAUR Italy
SRL IVA IT03841300167
Piazza Giacomo Matteotti, 6
24122 Bergamo Tel 02 36 00 65 93
Fax 02 36 00 65 94
italia@gentaur.com | Gentaur | Gentaur