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Information on EC 1.1.99.36 - alcohol dehydrogenase (nicotinoprotein)
for references in articles please use BRENDA:EC1.1.99.36
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EC Tree 1 Oxidoreductases
1.1 Acting on the CH-OH group of donors
1.1.99 With unknown physiological acceptors
1.1.99.36 alcohol dehydrogenase (nicotinoprotein)
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 .
The expected taxonomic range for this enzyme is: Bacteria, Archaea
- 1.1.99.36
-
nicotinoproteins
- erythropolis
-
rhodococcus
- dehydrogenases
- methanolica
- formaldehyde
- amycolatopsis
- nadp
- n,n-dimethyl-4-nitrosoaniline
- dismutase
- methanol
- synthesis
showSynonyms
ndma-adh, np-adh, ndma-dependent alcohol dehydrogenase, more
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
4-nitroso-N, N-dimethylaniline-dependent alcohol dehydrogenase
NDMA-ADH
NDMA-dependent alcohol dehydrogenase
4-nitroso-N, N-dimethylaniline-dependent alcohol dehydrogenase
-
-
-
NDMA-dependent alcohol dehydrogenase
-
-
-
-
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ethanol + acceptor = acetaldehyde + reduced acceptor
-
-
-
-
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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].
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
1-butanol + N,N-dimethyl-4-nitrosoaniline
butanaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
Substrates: -
Products: -
Products: -
?
1-hexanol + N,N-dimethyl-4-nitrosoaniline
hexanal + 4-(hydroxylamino)-N,N-dimethylaniline
Substrates: -
Products: -
Products: -
?
1-propanol + N,N-dimethyl-4-nitrosoaniline
propanaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
Substrates: -
Products: -
Products: -
?
2-butanol + N,N-dimethyl-4-nitrosoaniline
butanone + 4-(hydroxylamino)-N,N-dimethylaniline
Substrates: -
Products: -
Products: -
?
2-propanol + N,N-dimethyl-4-nitrosoaniline
acetone + 4-(hydroxylamino)-N,N-dimethylaniline
Substrates: -
Products: -
Products: -
?
acetate + N,N-dimethyl-4-nitrosoaniline
? + 4-(hydroxylamino)-N,N-dimethylaniline
Substrates: -
Products: -
Products: -
?
acetophenone + 1-propanol
?
-
Substrates: 4.3% yield, enantiomeric excess of 0.99 for (S)-product chiral secondary alcohol
Products: 3.8% yield, enantiomeric excess of 0.99 for (S)-product
Products: 3.8% yield, enantiomeric excess of 0.99 for (S)-product
r
acetophenone + cyclohexanol
?
-
Substrates: 2.8-3.7% yield, depending on the ratio of substrates, enantiomeric excess of 0.95-0.99 for (S)-product chiral secondary alcohol
Products: 3.8% yield, enantiomeric excess of 0.99 for (S)-product
Products: 3.8% yield, enantiomeric excess of 0.99 for (S)-product
r
acetophenone + ethanol
?
-
Substrates: 3.8% yield, enantiomeric excess of 0.99 for (S)-product chiral secondary alcohol
Products: 3.8% yield, enantiomeric excess of 0.99 for (S)-product
Products: 3.8% yield, enantiomeric excess of 0.99 for (S)-product
r
ethanol + N,N-dimethyl-4-nitrosoaniline
acetaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
methanol + N,N-dimethyl-4-nitrosoaniline
formaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
1-butanol + N,N-dimethyl-4-nitrosoaniline
butanal + 4-(hydroxylamino)-N,N-dimethylaniline
Substrates: -
Products: -
Products: -
?
1-butanol + N,N-dimethyl-4-nitrosoaniline
butanal + 4-(hydroxylamino)-N,N-dimethylaniline
-
Substrates: -
Products: -
Products: -
?
ethanol + N,N-dimethyl-4-nitrosoaniline
acetaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
Substrates: -
Products: -
Products: -
?
ethanol + N,N-dimethyl-4-nitrosoaniline
acetaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
-
Substrates: -
Products: -
Products: -
?
methanol + N,N-dimethyl-4-nitrosoaniline
formaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
Substrates: -
Products: -
Products: -
?
methanol + N,N-dimethyl-4-nitrosoaniline
formaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
Substrates: best substrate
Products: -
Products: -
?
methanol + N,N-dimethyl-4-nitrosoaniline
formaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
-
Substrates: -
Products: -
Products: -
?
methanol + N,N-dimethyl-4-nitrosoaniline
formaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
-
Substrates: best substrate
Products: -
Products: -
?
additional information
?
-
-
Substrates: enzyme shows formaldehyde dismutase activity producing methylformate when incubated with formaldehyde and methanol
Products: -
Products: -
?
additional information
?
-
Substrates: enzyme shows formaldehyde dismutase activity producing methylformate when incubated with formaldehyde and methanol
Products: -
Products: -
?
additional information
?
-
-
Substrates: no activity with electron acceptors NAD+, NADP+, 3-acetylpyridine adenine dinucleotide, thionicotinamide adenine dinucleotide, phenazine methosulfate, Wurster's Blue, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliurn bromide, or cytochrome c
Products: -
Products: -
?
additional information
?
-
Substrates: no activity with electron acceptors NAD+, NADP+, 3-acetylpyridine adenine dinucleotide, thionicotinamide adenine dinucleotide, phenazine methosulfate, Wurster's Blue, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliurn bromide, or cytochrome c
Products: -
Products: -
?
additional information
?
-
-
Substrates: no activity with electron acceptors NAD+, NADP+, 3-acetylpyridine adenine dinucleotide, thionicotinamide adenine dinucleotide, phenazine methosulfate, Wurster's Blue, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliurn bromide, or cytochrome c
Products: -
Products: -
?
additional information
?
-
-
Substrates: catalyzes the asymmetric reduction of ketones using cheap reductants, such as ethanol, with high stereoselectivity, but the reaction is too slow to obtain good yields
Products: -
Products: -
?
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ethanol + N,N-dimethyl-4-nitrosoaniline
acetaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
methanol + N,N-dimethyl-4-nitrosoaniline
formaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
ethanol + N,N-dimethyl-4-nitrosoaniline
acetaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
Substrates: -
Products: -
Products: -
?
ethanol + N,N-dimethyl-4-nitrosoaniline
acetaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
-
Substrates: -
Products: -
Products: -
?
methanol + N,N-dimethyl-4-nitrosoaniline
formaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
Substrates: -
Products: -
Products: -
?
methanol + N,N-dimethyl-4-nitrosoaniline
formaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
-
Substrates: -
Products: -
Products: -
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
NADH
comparison of NADH binding in nicotinoprotein alcohol dehydrogenase and in the conventional alcohol dehydrogenase. Considerable parts of the coenzyme binding in the conventional enzyme are also present in the nicotinoprotein. Alterations in two loop structures affect coenzyme binding and appear to tighten it in the nicotinoprotein model. The positions of residues in a sphere of 3.8 A around the coenzyme template of the nicotinoprotein model reveal the presence of coenzyme interactions additional to those in the conventional structure
NADH
The NADH absorbance spectrum of nicotinoprotein alcohol dehydrogenase has a maximum at 326 nm. Reduced enzyme-bound pyridine dinucleotide can be reversibly oxidized by acetaldehyde. The fluorescence excitation spectrum for NADH bound to the enzyme has a maximum at 325 nm. Upon excitation at 290 nm, energy transfer from tryptophan to enzyme-bound NADH is negligible. The fluorescence emission spectrum upon excitation at 325 nm for NADH bound to the enzyme has a maximum at 422 nm. The fluorescence lifetime of NADH bound to the nicotinoprotein is very short compared to enzyme-bound NADH complexes, also compared to NADH bound to horse liver alcohol dehydrogenase. The cofactor-protein interaction in the nicotinoprotein alcohol dehydrogenase active site is more rigid and apolar than that in horse liver alcohol dehydrogenase
NADH
tightly bound, but NDMA-dependent alcohol dehydrogenase is unable to use NAD(P)(H) as cofactors
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
acetaldehyde
1 mM, 40% inhibition; 40% inhibition at 1 mM, 60% at 2 mM, and 95% at 10 mM
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
additional information
-
NAD(H) and NADP(H) do neither inhibit nor stimulate NDMA-ADH activity; NAD(H) and NADP(H) neither inhibit nor stimulate NDMA-ADH activity
-
additional information
NAD(H) and NADP(H) do neither inhibit nor stimulate NDMA-ADH activity; NAD(H) and NADP(H) neither inhibit nor stimulate NDMA-ADH activity
-
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
-
NAD(H) and NADP(H) neither inhibit nor stimulate NDMA-ADH activity
-
additional information
NAD(H) and NADP(H) neither inhibit nor stimulate NDMA-ADH activity
-
additional information
-
no activation by phosphate and ammonium chloride. NAD(H) and NADP(H) do neither inhibit nor stimulate NDMA-ADH activity
-
additional information
no activation by phosphate and ammonium chloride. NAD(H) and NADP(H) do neither inhibit nor stimulate NDMA-ADH activity
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0016
trans-4-(N,N-dimethylamino)-cinnamaldehyde
uncompetitive inhibition term reflecting binding to the enzyme containing the oxidized coenzyme
0.0033
trans-4-(N,N-dimethylamino)-cinnamaldehyde
competitive inhibition term with respect to NDMA
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
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
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Highest Expressing Human Cell Lines
Cell Line Links Show AA Sequence and Transmembrane Helices (450 entries)
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
Please use the AA Sequence and Transmembrane Helices Search for a specific query.
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
110000
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
trimer
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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
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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
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
synthesis
-
enzyme catalyzes the asymmetric reduction of ketones using cheap reductants, such as ethanol, with high stereoselectivity, but the reaction is too slow to obtain good yields. For developing biotransformations of industrial interest using nicotinoprotein alcohol dehydrogenases, the attention should be focused on enzymes with a higher reactivity towards prochiral ketones and secondary alcohols
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Schenkels, P.; De Vries, S.; Straathof, A.J.J.
Scope and limitations of the use of nicotinoprotein alcohol dehydrogenase for the coenzyme-free production of enantiopure fine-chemicals
Biocatal. Biotransform.
19
191-212
2001
Rhodococcus erythropolis
-
brenda
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)
brenda
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)
brenda
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
brenda
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
brenda
EXTERNAL LINKS (specific for EC-Number 1.1.99.36)
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Release 2023.1 (January 2023)
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