Information on EC 1.2.1.46 - formaldehyde dehydrogenase

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

EC NUMBER
COMMENTARY hide
1.2.1.46
-
RECOMMENDED NAME
GeneOntology No.
formaldehyde dehydrogenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
formaldehyde + NAD+ + H2O = formate + NADH + 2 H+
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
redox reaction
reduction
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Chloroalkane and chloroalkene degradation
-
-
formaldehyde oxidation IV (thiol-independent)
-
-
Metabolic pathways
-
-
Methane metabolism
-
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methanol oxidation to carbon dioxide
-
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Microbial metabolism in diverse environments
-
-
SYSTEMATIC NAME
IUBMB Comments
formaldehyde:NAD+ oxidoreductase
-
CAS REGISTRY NUMBER
COMMENTARY hide
9028-84-6
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
P1 NCIB 11625
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain ZV 580
-
-
Manually annotated by BRENDA team
strain ZV 580
-
-
Manually annotated by BRENDA team
strain NRRL Y-11430, methylotrophic yeast
-
-
Manually annotated by BRENDA team
Komagataella pastoris NRRL Y-11430
strain NRRL Y-11430, methylotrophic yeast
-
-
Manually annotated by BRENDA team
Bath
-
-
Manually annotated by BRENDA team
Ogataea angusta Tf 11-6
strain Tf 11-6
-
-
Manually annotated by BRENDA team
C1
-
-
Manually annotated by BRENDA team
MS
-
-
Manually annotated by BRENDA team
Indian mackerel
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetaldehyde + NAD+ + H2O
acetate + NADH + H+
show the reaction diagram
aryl alcohol + NAD+ + H2O
? + NADH
show the reaction diagram
-
-
-
-
-
benzyl alcohol + NAD+ + H2O
? + NADH
show the reaction diagram
-
-
-
-
-
formaldehyde + H2O + NAD+
formate + NADH + H+
show the reaction diagram
formaldehyde + NAD+ + H2O
formate + NADH
show the reaction diagram
formaldehyde + NAD+ + H2O
formate + NADH + H+
show the reaction diagram
glyoxal + NAD+ + H2O
glyoxylate + NADH
show the reaction diagram
isobutanol + NAD+ + H2O
? + NADH
show the reaction diagram
-
-
-
-
-
isopentanol + NAD+ + H2O
? + NADH
show the reaction diagram
-
-
-
-
-
n-butanol + NAD+ + H2O
? + NADH
show the reaction diagram
-
-
-
-
-
n-hexanol + NAD+ + H2O
? + NADH
show the reaction diagram
-
-
-
-
-
n-pentanol + NAD+ + H2O
? + NADH
show the reaction diagram
-
-
-
-
-
n-propanol + NAD+ + H2O
? + NADH
show the reaction diagram
-
-
-
-
-
propionaldehyde + NAD+ + H2O
propionate + NADH
show the reaction diagram
pyruvaldehyde + NAD+ + H2O
pyruvate + NADH
show the reaction diagram
S-hydroxymethylglutathione + NAD+ + H2O
S-formylglutathione + NADH + H+
show the reaction diagram
-
-
-
-
?
S-nitrosoglutathione + NADH + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
formaldehyde + NAD+ + H2O
formate + NADH
show the reaction diagram
formaldehyde + NAD+ + H2O
formate + NADH + H+
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
glutathione
NADH
-
-
additional information
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
independent of Mg2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,2'-bipyridine
4-Methylpyrazole
-
native and recombinant protein
5,5'-dithiobis(2-nitrobenzoate)
-
partial
8-hydroxyquinoline
acetaldehyde
-
substrate n-butanol
diisopropylphosphofluoridate
-
partial
dodecanoic acid
-
-
ethanol
-
substrate n-butanol
Mn2+
-
partial
N-Acetylimidazole
-
partial
n-butanol
-
; substrate formaldehyde
n-Butyraldehyde
-
substrate formaldehyde or n-butanol
n-Hexanol
-
; substrate formaldehyde
n-Propanol
-
substrate n-butanol
n-Valeraldehyde
-
substrate n-butanol
o-phenanthroline
p-chloromercuribenzoate
-
-
phenylmethanesulfonyl fluoride
-
-
propionaldehyde
-
substrate formaldehyde
Sodium tetrathionate
-
partial
Zn2+
-
partial
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
methanol
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5.9
acetaldehyde
-
-
100
aryl alcohol
-
-
5.5
benzyl alcohol
-
-
0.067 - 119.9
formaldehyde
30
isobutanol
-
-
6.2
Isobutyraldehyde
-
-
4.2
Isopentanol
-
-
10.6
n-butanol
-
-
4
n-Hexanol
-
-
6.5
n-Pentanol
-
-
50
n-Propanol
-
-
0.0033 - 0.5
NAD+
0.002 - 0.011
NADH
47
propionaldehyde
-
-
0.0017 - 2.65
S-hydroxymethylglutathione
0.0048 - 0.16
S-nitrosoglutathione
additional information
additional information
-
Km for NAD+ with various aldehydes and alcohols
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.03
4-Methylpyrazole
-
native and recombinant protein
6.1
acetaldehyde
-
substrate n-butanol
0.148 - 0.617
dodecanoic acid
90
ethanol
-
substrate n-butanol
5.4
n-butanol
-
substrate formaldehyde
0.27 - 2.6
n-Butyraldehyde
2.3
n-Hexanol
-
substrate formaldehyde
70
n-Propanol
-
substrate n-butanol
1.9
n-Valeraldehyde
-
substrate n-butanol
35
propionaldehyde
-
substrate formaldehyde
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5
-
the stable recombinant strain Tf 11-6 in cell-free extract
7
-
FdDH specific activity in cell-free extract of Tf 11-6, cultivated in methanol medium supplemented with 10 mM formaldehyde
12
-
at 20C
17
-
3.8fold purified enzyme at 25C
24.1
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recombinant protein, expressed in Escherichia coli
27
-
3.8fold purified enzyme at 37C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.8
-
formaldehyde
8
-
Nafion membrane-immobilized enzyme
8.1 - 8.5
-
-
8.4
-
best sensitivity of the biosensor output signal is observed in 2.5 mM borate buffer, pH 8.4
8.9
-
formaldehyde
10.8
-
n-butanol
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.6 - 8.3
-
at pH 6.6 and 8.3 about 50% of activity maximum
7.5 - 8.5
-
-
7.6 - 8.3
-
optimal pH-value for the developed FdDH-based biosensor
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
S-nitrosoglutathione
30
-
S-hydroxymethylglutathione
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45 - 50
-
optimal temperature for the developed FdDH-based biosensor
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
43500
-
gel filtration
150000
250000
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
-
-
monomer
-
1 * 48000, SDS-PAGE
tetramer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
formaldehyde dehydrogenase-adenosine 5'-diphosphate ribose and E67L-NADH binary complexes are determined
-
hanging drop vapor diffusion method
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 8.5
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-
686818
8
-
15 h, 20% loss of activity
288248
8.5 - 10
-
stable
288248
additional information
-
FDH is not stable at a pH below 6.5, but shows higher stability in an alkaline pH domain
690067
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 65
-
at 65C, the enzyme retains about 60% of its highest activity (assay time of ca. 5 min), which is equal to the level of FdDH activity at 30C
40
-
30 min, pH 7.5, stable
45
-
stable for 6 min
55
-
pH 7.5, 30 min, 50% loss of activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-10C, ammonium sulfate solution (80% saturation), 2 mM DTT
-
-20C, stable for at least 1 month
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14 h and 1 mM during 22 h, the storage stability of the developed biosensors is found to be longer than 18 days at 4C
-
4C, Nafion membrane-immobilized enzyme, more than 6 months, remains stable
-
the developed sensors demonstrate a good operational stability at comparatively low formaldehyde concentrations of 2 mM during
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; recombinant protein, expressed in Escherichia coli
-
by ion-exchange chromatography
-
by two-step anion-exchange chromatography and precipitation by 80% (NH4)2SO4, precipitate, 3.8fold purified
-
DE52, Q-sepharose and Sephacryl S-100HR chromatographic steps
-
FdDH ss isolated from the cell-free extract by a two-step ion exchange chromatography
-
partially purified
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
construction of a recombinant yeast clone originated from the recipient strain of thermotolerant methylotrophic yeast Hansenula polymorpha which is a NAD+- and glutathione dependent FdDH over-producer
-
expressed in Escherichia coli
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FLD1 gene with its own promoter is recloned into LEU2-containing integrative plasmid pYT1 devoid of the autonomously replicating sequence to be used for multicopy integration of the gene into chromosome of the recipient strain NCYC 495
-
FLD1 gene with the own promoter inserted into the integrative plasmid pYT1 containing LEU2 gene of Saccharomyces cerevisiae (as a selective marker). The constructed vector is used for multi-copy integration of the target gene into genome by transformation of the recipient cells of the strain NCYC 495
-
for expression in Escherichia coli BL21 cells
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E67L
-
mutant shows, that Glu67 is critical for capturing the substrates for catalysis
R368L
-
mutant shows, that the predominant role of Arg-368 is in the binding of the coenzyme
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
biotechnology
-
development of novel formaldehyde-selective amperometric biosensors based on immobilized NAD+- and glutathione dependent formaldehyde dehydrogenase with high selectivity to formaldehyde and a low cross-sensitivity to other substances, the laboratory prototype of the sensor is applied for FA testing in some real samples of pharmaceutical (formidron), disinfectant (descoton forte) and industrial product (formalin)
degradation
industry
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