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Information on EC 1.1.99.36 - alcohol dehydrogenase (nicotinoprotein) and Organism(s) Amycolatopsis methanolica and UniProt Accession P80175

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IUBMB Comments
Contains Zn2+. Nicotinoprotein alcohol dehydrogenases are unique medium-chain dehydrogenases/reductases (MDR) alcohol dehydrogenases that have a tightly bound NAD+/NADH cofactor that does not dissociate during the catalytic process. Instead, the cofactor is regenerated by a second substrate or electron carrier. While the in vivo electron acceptor is not known, N,N-dimethyl-4-nitrosoaniline (NDMA), which is reduced to 4-(hydroxylamino)-N,N-dimethylaniline, can serve this function in vitro. The enzyme from the Gram-positive bacterium Amycolatopsis methanolica can accept many primary alcohols as substrates, including benzylalcohol .
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Amycolatopsis methanolica
UNIPROT: P80175
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The taxonomic range for the selected organisms is: Amycolatopsis methanolica
The expected taxonomic range for this enzyme is: Bacteria, Archaea
Synonyms
ndma-adh, np-adh, ndma-dependent alcohol dehydrogenase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4-nitroso-N, N-dimethylaniline-dependent alcohol dehydrogenase
-
-
NDMA-dependent alcohol dehydrogenase
SYSTEMATIC NAME
IUBMB Comments
ethanol:acceptor oxidoreductase
Contains Zn2+. Nicotinoprotein alcohol dehydrogenases are unique medium-chain dehydrogenases/reductases (MDR) alcohol dehydrogenases that have a tightly bound NAD+/NADH cofactor that does not dissociate during the catalytic process. Instead, the cofactor is regenerated by a second substrate or electron carrier. While the in vivo electron acceptor is not known, N,N-dimethyl-4-nitrosoaniline (NDMA), which is reduced to 4-(hydroxylamino)-N,N-dimethylaniline, can serve this function in vitro. The enzyme from the Gram-positive bacterium Amycolatopsis methanolica can accept many primary alcohols as substrates, including benzylalcohol [1].
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1-butanol + N,N-dimethyl-4-nitrosoaniline
butanaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
show the reaction diagram
-
-
-
?
1-propanol + N,N-dimethyl-4-nitrosoaniline
propanaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
show the reaction diagram
-
-
-
?
2-butanol + N,N-dimethyl-4-nitrosoaniline
butanone + 4-(hydroxylamino)-N,N-dimethylaniline
show the reaction diagram
-
-
-
?
2-propanol + N,N-dimethyl-4-nitrosoaniline
acetone + 4-(hydroxylamino)-N,N-dimethylaniline
show the reaction diagram
-
-
-
?
ethanol + N,N-dimethyl-4-nitrosoaniline
acetaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
show the reaction diagram
methanol + N,N-dimethyl-4-nitrosoaniline
formaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
show the reaction diagram
-
-
-
?
1-butanol + N,N-dimethyl-4-nitrosoaniline
butanal + 4-(hydroxylamino)-N,N-dimethylaniline
show the reaction diagram
-
-
-
?
1-hexanol + N,N-dimethyl-4-nitrosoaniline
hexanal + 4-(hydroxylamino)-N,N-dimethylaniline
show the reaction diagram
-
-
-
?
acetate + N,N-dimethyl-4-nitrosoaniline
? + 4-(hydroxylamino)-N,N-dimethylaniline
show the reaction diagram
-
-
-
?
ethanol + N,N-dimethyl-4-nitrosoaniline
acetaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
show the reaction diagram
-
-
-
?
methanol + N,N-dimethyl-4-nitrosoaniline
formaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ethanol + N,N-dimethyl-4-nitrosoaniline
acetaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
show the reaction diagram
-
-
-
?
methanol + N,N-dimethyl-4-nitrosoaniline
formaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
show the reaction diagram
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADH
tightly bound, but NDMA-dependent alcohol dehydrogenase is unable to use NAD(P)(H) as cofactors
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Zn2+
takes part in catalysis
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-butanol
substrate inhibition
1-propanol
substrate inhibition
2-butanol
substrate inhibition
2-propanol
substrate inhibition
acetaldehyde
1 mM, 40% inhibition
Cu2+
1 mM, 90% inhibition
ethanol
substrate inhibition
Fe2+
1 mM, 90% inhibition
Isobutyramide
competitive to N,N-dimethyl-4-nitrosoaniline
KCN
2 mM, 90% inhibition
NaN3
2 mM, 25% inhibition
trans-4-(N,N-dimethylamino)-cinnamaldehyde
inhibition through direct binding to the catalytic zinc ion in a substrate-like geometry. This binding is accompanied by a characteristic red shift of the aldehyde absorbance from 398 nm to 467 nm
trifluoroethanol
nonreactive substrate analogue, competitive to ethanol
Zn2+
1 mM, 40% inhibition
acetaldehyde
40% inhibition at 1 mM, 60% at 2 mM, and 95% at 10 mM
ADP
10% inhibition at 1 mM, 50% at 10 mM
AMP
10% inhibition at 1 mM, 50% at 10 mM
ATP
10% inhibition at 1 mM, 50% at 10 mM
Cu2+
90% inhibition at 1 mM
Fe2+
90% inhibition at 1 mM
Hg2+
100% inhibition at 1 mM
KCN
90% inhibition at 2 mM
NaN3
25% inhibition at 2 mM
Ni2+
15% inhibition at 1 mM
Zn2+
40% inhibition at 1 mM
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0025
1-butanol
pH 7.0, 45°C
0.0039
1-propanol
pH 7.0, 45°C
10.1
2-butanol
pH 7.0, 45°C
13.4
2-propanol
pH 7.0, 45°C
0.082
ethanol
pH 7.0, 45°C
additional information
additional information
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.65
1-butanol
pH 7.0, 45°C
4
1-propanol
pH 7.0, 45°C
4.4
2-butanol
pH 7.0, 45°C
14.5
2-propanol
pH 7.0, 45°C
33
ethanol
pH 7.0, 45°C
0.046
Isobutyramide
pH 7.0, 20°C
0.0016 - 0.0033
trans-4-(N,N-dimethylamino)-cinnamaldehyde
0.0016
trifluoroethanol
pH 7.0, 20°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.003
substrate methanol, pH 7.0, 45°C, extracts of cells grown on ethanol
0.004
substrate ethanol, pH 7.0, 45°C, extracts of cells grown on ethanol
0.005
substrate methanol, pH 7.0, 45°C, extracts of cells grown in fed-batch culture on methanol
0.018
substrate ethanol, pH 7.0, 45°C, extracts of cells grown in fed-batch culture on methanol
3.1
45°C, pH 7.0, purified enzyme
3.1
purified enzyme, pH 7.0, 45°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
ADHN_AMYME
371
0
38992
Swiss-Prot
-
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
120000
gel filtration
38970
x * 38970, calculated
39000
3 * 39000, SDS-PAGE
39000
3 * 39000, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 38970, calculated
trimer
3 * 39000, SDS-PAGE
trimer
3 * 39000, SDS-PAGE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
docking study of trans-4-(N,N-dimethylamino)-cinnamaldehyde to the enzyme model active site. The np-ADH model accommodates the inhibitor in a substrate-like conformation without collision with inner-sphere and secondary sphere residues
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
native enzyme 163fold to homogeneity by ammonium sulfate fractionation, hydrophobic interaction chromatography, ultrafiltration, anion exchange and hydroxyapatite chromatography, followed by gel filtration
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Piersma, S.R.; Visser, A.J.; de Vries, S.; Duine, J.A.
Optical spectroscopy of nicotinoprotein alcohol dehydrogenase from Amycolatopsis methanolica: a comparison with horse liver alcohol dehydrogenase and UDP-galactose epimerase
Biochemistry
37
3068-3077
1998
Amycolatopsis methanolica (P80175)
Manually annotated by BRENDA team
Norin, A.; Piersma, S.R.; Duine, J.A.; Jornvall, H.
Nicotinoprotein (NAD+ -containing) alcohol dehydrogenase: structural relationships and functional interpretations
Cell. Mol. Life Sci.
60
999-1006
2003
Amycolatopsis methanolica (P80175)
Manually annotated by BRENDA team
Van Ophem, P.W.; Van Beeumen, J.; Duine, J.A.
Nicotinoprotein [NAD(P)-containing] alcohol/aldehyde oxidoreductases. Purification and characterization of a novel type from Amycolatopsis methanolica
Eur. J. Biochem.
212
819-826
1993
Amycolatopsis methanolica, Amycolatopsis methanolica (P80175), Amycolatopsis methanolica NCIB 11946
Manually annotated by BRENDA team
Piersma, S.R.; Norin, A.; de Vries, S.; Jornvall, H.; Duine, J.A.
Inhibition of nicotinoprotein (NAD+-containing) alcohol dehydrogenase by trans-4-(N,N-dimethylamino)-cinnamaldehyde binding to the active site
J. Protein Chem.
22
457-461
2003
Amycolatopsis methanolica (P80175), Amycolatopsis methanolica
Manually annotated by BRENDA team