Information on EC 1.6.1.1 - NAD(P)+ transhydrogenase (Si-specific)

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY
1.6.1.1
-
RECOMMENDED NAME
GeneOntology No.
NAD(P)+ transhydrogenase (Si-specific)
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
NADPH + NAD+ = NADP+ + NADH
show the reaction diagram
mechanism
-
NADPH + NAD+ = NADP+ + NADH
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
NAD/NADH phosphorylation and dephosphorylation
-
-
NAD metabolism
-
-
Nicotinate and nicotinamide metabolism
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
NADPH:NAD+ oxidoreductase (Si-specific)
The enzyme from Azotobacter vinelandii is a flavoprotein (FAD). It is Si-specific with respect to both NAD+ and NADP+. Also acts on deamino coenzymes [cf. EC 1.6.1.2 NAD(P)+ transhydrogenase (Re/Si-specific)].
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
energy-independent soluble pyridine nucleotide transhydrogenase
-
H+-thase
-
-
-
-
NAD transhydrogenase
-
-
-
-
NAD(P) transhydrogenase
-
-
-
-
NAD(P)(+) transhydrogenase [B-specific]
-
-
-
-
NADH transhydrogenase
-
-
-
-
NADH-NADP-transhydrogenase
-
-
-
-
NADPH-NAD oxidoreductase
-
-
-
-
NADPH-NAD transhydrogenase
-
-
-
-
NADPH:NAD+ transhydrogenase
-
-
-
-
nicotinamide adenine dinucleotide (phosphate) transhydrogenase
-
-
-
-
nicotinamide nucleotide transhydrogenase
-
-
-
-
non-energy-linked transhydrogenase
-
-
-
-
pyridine nucleotide transferase
-
-
-
-
pyridine nucleotide transhydrogenase
-
-
-
-
soluble pyridine nucleotide transhydrogenase
-
-
soluble pyridine nucleotide transhydrogenase
-
soluble pyridine nucleotide transhydrogenase
Escherichia coli JM109
-
-
-
soluble transhydrogenase
-
-
soluble transhydrogenase
Escherichia coli JM109
-
;
-
STH
-
-
-
-
STH
Escherichia coli JM109
-
-
-
transhydrogenase
-
-
-
-
transhydrogenase, nicotinamide adenine dinucleotide (phosphate)
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9014-18-0
not distinguished from EC 1.6.1.2
9072-60-0
not distinguished from EC 1.6.1.2
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Azotobacter agilis
-
-
-
Manually annotated by BRENDA team
Chromatium sp.
-
-
-
Manually annotated by BRENDA team
substrain MG1655, gene sth
UniProt
Manually annotated by BRENDA team
potato tubers
-
-
Manually annotated by BRENDA team
formerly Pseudomonas natriegens
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
two pyridine nucleotide transhydrogenases: the energy-independent soluble pyridine nucleotide transhydrogenase (STH or UdhA) (EC 1.6.1.1) and the membrane-bound, energy-dependent pyridine nucleotide transhydrogenase (TH or PntAB) (EC 1.6.1.2). PntAB is widely distributed in the mitochondria and some bacteria, while STH is found only in certain Gammaproteobacteria and gram-positive bacteria
physiological function
the soluble pyridine nucleotide transhydrogenase, STH, is an energy-independent flavoprotein that directly catalyzes hydride transfer between NAD(H) and NADP(H) to maintain homeostasis of these two redox cofactors
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
NAD(P)H + 2,6-dichlorophenolindophenol
NAD(P)+ + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
-
-
-
?
NAD(P)H + K4Fe(CN)6
NAD(P)+ + K3Fe(CN)6
show the reaction diagram
-
-
-
?
NADH + 2'-NADP+
NAD+ + 2'-NADPH
show the reaction diagram
-
-
-
?
NADH + 3'-NADP+
NAD+ + 3'-NADPH
show the reaction diagram
-
-
-
?
NADH + NADP+
NADPH + NAD+
show the reaction diagram
-
-
-
?
NADH + thio-NAD+
NAD+ + thio-NADH
show the reaction diagram
-
-
-
?
NADH + thio-NADP+
NAD+ + thio-NADPH
show the reaction diagram
-
-
-
r
NADH + thio-NADP+
NAD+ + thio-NADPH
show the reaction diagram
-
-
-
?
NADH + thio-NADP+
NAD+ + thio-NADPH
show the reaction diagram
Escherichia coli JM109
-
-
-
r
NADP+ + NADH
NADPH + NAD+
show the reaction diagram
-
-
-
r
NADP+ + NADH
NADPH + NAD+
show the reaction diagram
-
-
-
-
NADP+ + NADH
NADPH + NAD+
show the reaction diagram
-
-
-
r
NADP+ + NADH
NADPH + NAD+
show the reaction diagram
-
-
-
-
NADP+ + NADH
NADPH + NAD+
show the reaction diagram
-
-
-
r
NADP+ + NADH
NADPH + NAD+
show the reaction diagram
-
4B-specific for NAD(P)H
-
?
NADP+ + NADH
NADPH + NAD+
show the reaction diagram
-
4B-specific for NAD(P)H
-
-
NADP+ + NADH
NADPH + NAD+
show the reaction diagram
-
reduction of NADP+ is preferred
-
r
NADP+ + NADH
NADPH + NAD+
show the reaction diagram
-
reduction of NADP+ is preferred
-
r
NADP+ + NADH
NADPH + NAD+
show the reaction diagram
Chromatium sp.
-
poorly reversible
-
r
NADP+ + NADH
NADPH + NAD+
show the reaction diagram
-
diaphorase-type reactions with NAD(P)H, K3Fe(CN)6 and 2,6-dichlorophenol indophenol
-
-
NADP+ + NADH
NADPH + NAD+
show the reaction diagram
-
diaphorase-type reactions with NAD(P)H, K3Fe(CN)6 and 2,6-dichlorophenol indophenol
-
-
NADP+ + NADH
NADPH + NAD+
show the reaction diagram
-
degree of reversibility depends on source of enzyme
-
r
NADP+ + NADH
NADPH + NAD+
show the reaction diagram
Escherichia coli JM109
-
-
-
r
NADPH + 3-acetylpyridine-NAD+
NADP+ + 3-acetylpyridine-NADH
show the reaction diagram
-
-
-
?
NADPH + 3-acetylpyridine-NAD+
NADP+ + 3-acetylpyridine-NADH
show the reaction diagram
-
-
-
?
NADPH + 3-acetylpyridine-NAD+
NADP+ + 3-acetylpyridine-NADH
show the reaction diagram
-
-
-
?
NADPH + deamino-NAD+
NADP+ + deamino-NADH
show the reaction diagram
-
-
-
?
NADPH + deamino-NAD+
NADP+ + deamino-NADH
show the reaction diagram
-
-
-
?
NADPH + deamino-NAD+
NADP+ + deamino-NADH
show the reaction diagram
-
-
-
?
NADPH + deamino-NAD+
NADP+ + deamino-NADH
show the reaction diagram
-
-
-
?
NADPH + deamino-NAD+
NADP+ + deamino-NADH
show the reaction diagram
-
-
-
?
NADPH + NAD+
NADP+ + NADH
show the reaction diagram
-
-
r
NADPH + pyridine aldehyde-NAD+
NADP+ + pyridine aldehyde-NADH
show the reaction diagram
-
-
-
?
NADPH + pyridine aldehyde-NAD+
NADP+ + pyridine aldehyde-NADH
show the reaction diagram
-
-
-
?
NADPH + pyridine aldehyde-NAD+
NADP+ + pyridine aldehyde-NADH
show the reaction diagram
-
-
-
?
NADPH + thio-NAD+
NADP+ + thio-NADH
show the reaction diagram
-
-
-
?
NADPH + thio-NAD+
NADP+ + thio-NADH
show the reaction diagram
EcSTH has a 1.25fold preference for NADPH over thio-NAD+
-
r
NADPH + thio-NADP+
NADP+ + thio-NADPH
show the reaction diagram
-
-
-
?
NADPH + thio-NADP+
NADP+ + thio-NADPH
show the reaction diagram
-
-
-
?
NADPH + thio-NADP+
NADP+ + thio-NADPH
show the reaction diagram
-
-
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
NADH + NADP+
NADPH + NAD+
show the reaction diagram
-
-
-
?
NADPH + NAD+
NADP+ + NADH
show the reaction diagram
P27306
-
-
r
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
FAD
-
reduction of FAD leads to its dissociation at enzyme concentrations of 10-100 nM and 30C-40C, the apoenzyme can be reactivated to 10-15% by addition of FAD
FAD
-
FAD dissociates from the enzyme by heat treatment at 100C or by treatment with 5% trichloroacetic acid at 0C
FAD
-
inactivation by heat treatment can be reversed by addition of FAD
flavin
flavoprotein
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Ca2+
-
10-20fold activation of NAD+ reduction by NADPH at alkaline pH in cell-free extracts
Ca2+
-
Ca2+-dependent allosteric conformational change, competitive inhibition of activation by Mg2+
EDTA
-
slight activating at low buffer concentrations
K+
-
no full activation compared to Ca2+
Mg2+
-
10-20fold activation of NAD+ reduction by NADPH at alkaline pH in cell-free extracts
Mg2+
-
no full activation even at saturation concentrations
Mn2+
-
compared to Ca2+ 10 times higher concetrations are required to obtain the same degree of activation
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2'-AMP
Chromatium sp.
-
-
2-mercaptoethanol
-
5'-AMP
-
5 mM, 92% inhibition of NADP+ reduction, complete inhibition of NAD+ reduction
ADP
-
5 mM, 94% inhibition of NADP+ reduction, complete inhibition of NAD+ reduction
arsenate
-
complete inhibition of activity in either direction
ATP
-
5 mM, 81% inhibition of NADP+ reduction, complete inhibition of NAD+ reduction
Ca2+
slightly inhibitory
CTP
-
5 mM, 86% inhibition of NADP+ reduction
deoxycholate
-
-
diphosphate
-
5 mM, 91% inhibition of NADP+ reduction
DMSO
slightly inhibitory
GTP
-
5 mM, 89% inhibition of NADP+ reduction
NAD+
-
competitive to thio-NAD+, uncompetitive with respect to NADPH
NADP+
-
0.01 mM, 28% inhibition of NAD+ reduction with NADPH, 12.5% inhibition of NADP+ reduction with NADPH; inhibition of 2'-AMP activated reaction
NADP+
-
inhibition in absence of Ca2+; strong inhibition in the absence of Ca2+, saturation with Ca2+ completely abolishes inhibition
NADP+
-
uncompetitive to thio-NAD+
NADPH
EcSTH activity is strongly inhibited by excess NADPH, but not by thio-NAD+
p-Aminophenylarsenoxide
-
0.1 mM, 40-60% inhibition in the absence of either phosphate or magnesium ions, reduction of NAD+ by NADPH in cell-free extracts is rapidly and completely inhibited in the presence of 20 mM phosphate
p-chloromercuribenzoate
-
0.044 mM, 40-50% inhibition after 30 min, activity can be restored by adding 2-mercaptoethanol
p-hydroxymercuribenzoate
-
-
p-hydroxymercuribenzoate
-
-
p-hydroxymercuribenzoate
-
dependent on presence of oxidized or reduced substrate
phosphate
-
10-25 mM, 60-70% inhibition of purified enzyme, complete inhibition of enzyme in cell-free extracts by 5-10 mM phosphate, NADP+ reduction by NADH is inhibited, reduction of NAD+ by NADPH is hardly affected
phosphate
-
5 mM, complete inhibition of activity in either direction
phosphoenolpyruvate
-
87% inhibition of NADP+ reduction
pyridoxal 5'-phosphate
-
5 mM, 91% inhibition of NADP+ reduction
TTP
-
2 mM, 71% inhibition of NADP+ reduction
additional information
-
not inhibited by palmitoyl-CoA, not affected by treatment with 0.2 mg trypsin/mg protein
-
additional information
no or poor inhibition of the enzyme by Na+, Rb+, K+, Li+,, and Mg2+
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2'-AMP
-
0.5 mM, 26% activation of NAD+ reduction by NADPH, 12.5% activation of NADP+ reduction by NADPH, 1100% activation of NAD+ reduction by NADH, 21% activation of NADP+ reduction by NADH
2'-AMP
-
activation of NADP+ reduction by NADH is strongly Ca2+ dependent at nonsaturating concentrations of 2'-AMP; partially replaceable by 0.3 mM 2',3'-cyclic AMP or coenzyme A
2'-AMP
-
almost no effect
2'-AMP
-
activation of reduction of NADP+ or thio-NAD+ by NADH, no effect on reduction of NAD+ or thio-NAD+ by NADPH
2'-AMP
-
-
ADP
75.5% activation at 2 mM
AMP
71.8% activation at 2 mM
ATP
53.4% activation at 2 mM
p-hydroxymercuribenzoate
-
activation, depending on presence of oxidized or reduced substrates
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.4
deamino-NAD+
-
cosubstrate NADPH
0.0003
FAD
-
at 25C
0.11
NAD+
-
cosubstrate NADPH
0.38
NAD+
-
cosubstrate NADPH
0.025
NADH
-
cosubstrate NADP+
0.06
NADH
-
+ thio-NAD+
0.077
NADH
-
+ thio-NAD+
0.085
NADH
-
+ thio-NADP+
0.015
NADPH
-
cosubstrate NAD+
0.0683
NADPH
pH 7.5, 35C
0.04
thio-NAD+
-
NADPH + thio-NAD+
0.04
thio-NAD+
-
cosubstrate NADH
0.05
thio-NAD+
-
cosubstrate NADH
0.075
thio-NAD+
-
cosubstrate NADPH
0.1332
thio-NAD+
pH 7.5, 35C
0.25
thio-NAD+
-
cosubstrate NADPH
0.03
thio-NADP+
-
cosubstrate NADH
0.0025
FAD
-
at 0C
additional information
additional information
-
kinetic studies
-
additional information
additional information
-
dependency on Mg2+ concentration
-
additional information
additional information
-
kinetic studies
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
167.9
NADPH
Escherichia coli
P27306
pH 7.5, 35C
233 - 333
NADPH
Pseudomonas aeruginosa
-
-
259.5
thio-NAD+
Escherichia coli
P27306
pH 7.5, 35C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.0094
-
activity in inside-out mitochondrial particles from leaf
0.0131
-
activity in inside-out mitochondrial particles from tuber
24
Chromatium sp.
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7
-
NADH formation
7
-
around pH 7.0
9.6
-
NADPH formation
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7 - 10.5
-
pH 7.0: about 55% of maximal activity, pH 10.5: about 45% of maximal activity
8.4 - 8.7
-
half maximal activity in this pH range in the absence of Ca2+ or Mg2+, no activity above
9.1 - 9.3
-
half maximal activity in this pH range in the presence of 5 mM Ca2+ or Mg2+, no activity above
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
inside-out submitochondrial particles
Manually annotated by BRENDA team
Escherichia coli JM109
-
-
-
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
53000
-
SDS-PAGE
696915
421000
-
purified enzyme: exceptional stability of polymers at neutral pH
392577
421000
-
octameric form at pH 8.5-9.0, sedimentation equilibrium
392578
1600000
-
sedimentation equilibrium in presence of 2'-AMP
392582
6400000
-
sedimentation equilibrium in presence of NADP+
392582
27000000
-
rod-like polymers, electron micrography
392587
30000000 - 50000000
-
rod-like structure, light scattering
392586
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
octamer
-
x * 52000, model of quarternary structure, SDS-PAGE, immunoblot
octamer
-
8 * 54000, four subunits are placed in a rhomb, a second tetramer is located staggered on top of the first one, deduced from amino acid analysis and electron micrography of purified enzyme; x * 52000, SDS-PAGE, immunoblot
octamer
-
octameric in cell-free extract, polymeric in purified form
octamer
-
8 * 54000, SDS-PAGE
octamer
-
x * 54000, SDS-PAGE
octamer
-
x * 52000, SDS-PAGE, immunoblot; x * 58000, SDS-PAGE
octamer
-
x * 54000, SDS-PAGE
octamer
Azotobacter vinelandii ATCC478
-
8 * 54000, four subunits are placed in a rhomb, a second tetramer is located staggered on top of the first one, deduced from amino acid analysis and electron micrography of purified enzyme; x * 52000, SDS-PAGE, immunoblot
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7
-
polymeric enzyme stable
392577
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
50
-
1 h stable, inactivation is dramatically accelerated by NADH and NADPH, partial protection by NADP+ and FMN, almost full protection by FAD
392586
50
purified enzyme, pH 7.5, 5h, 50% activity remaining, 80% activity remaining after 30 min
725040
51
-
25 min, 50% inactivation, accelerated by addition of NADPH, reactivation by FAD
392590
55
-
approx. 50% activity lost after about 2 min, almost complete loss of activity after 20 min, biphasic inactivation: 70% activity lost with a first-order inactivation constant, 30% is lost much more rapidly, rate of thermal inactivation depends on concentration of NAD+, NADP+, NADH, NADPH, free FAD, Mg2+ and phosphate, independent of pH between pH 5 and pH 9, significant acceleration outside this range, addition of 1 mM FAD lowers inactivation rate about 20fold
392575
57
purified enzyme, pH 7.5, 30 min, 50% activity remaining
725040
62
purified enzyme, pH 7.5, 30 min, 10% activity remaining
725040
65
-
15 min, complete inactivation, protection by FAD
392586
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
50% inactivation after 10 min in 1 M guanidinium HCl
-
bovine serum albumin, 0.2%, stabilization of diluted solutions
-
urea, 5 min, 50% inactivation
-
urea, 8 M, no dissociation, complete inactivation of enzyme activity in 5 M urea
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4C, 0.1 M phosphate buffer, pH 7.5, 1 mM EDTA, several months, no loss of activity, storage at -20C yields a partly insoluble enzyme
-
25C, purified enzyme, 25 days, 65% activity remaining
4C, purified enzyme, 25 days, stable
-20C, 50 mM Tris-HCl, pH 7.0, 2 mM dithiothreitol, several weeks, no loss of activity, great loss of activity after several months
-
-15C, 50 mM Tris-HCl buffer, pH 7.5, no loss of activity in 12 weeks
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
affinity chromatography
-
affinity chromatography on immobilized 2'-AMP
-
affinity chromatography, native and recombinant enzyme
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Escherichia coli strain is transformed with a two plasmid system, one encoding the udhA gene and the other one encoding the phb operon
-
expression of the soluble enzyme in Escherichia coli strain DH5alpha
expression in Escherichia coli
-
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
inactivation by 4 M guanidinium HCl, 4-6% reactivation after transfer in 100 mM Tris buffer, pH 7.5 containing 100 mM FAD
-
urea inactivation: 3% reactivation after dialysis against buffer containing 0.1 mM FAD, 35% after dialysis against buffer containing 1% mercaptoethanol and 95% reactivation after dialysis against buffer containing both 0.1 mM FAD and 1% mercaptoethanol
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
biotechnology
-
enzymatic NADH production system in reverse micelles using a bacterial glycerol dehydrogenase. The present system is further extended to NADPH production in reverse micelles by coupling with a bacterial soluble transhydrogenase that catalyses the conversion of NADP+ to NADPH using NADH. Glycerol dehydrogenase and soluble transhydrogenase have potential for use in redox cofactor recycling in reverse micelles, which allows the use of catalytic quantities of NAD(P)H in organic media
biotechnology
-
Escherichia coli strain is transformed with a two plasmid system, one encoding the udhA gene and the other one encoding the phb operon. The functionality of this particular system is successfully demonstrated in PHB production experiments. Both productivity and yield of PHB can be increased when NADPH availability is increased
biotechnology
Escherichia coli JM109
-
enzymatic NADH production system in reverse micelles using a bacterial glycerol dehydrogenase. The present system is further extended to NADPH production in reverse micelles by coupling with a bacterial soluble transhydrogenase that catalyses the conversion of NADP+ to NADPH using NADH. Glycerol dehydrogenase and soluble transhydrogenase have potential for use in redox cofactor recycling in reverse micelles, which allows the use of catalytic quantities of NAD(P)H in organic media
-