Information on EC 1.1.1.244 - methanol dehydrogenase

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The expected taxonomic range for this enzyme is: Bacteria

EC NUMBER
COMMENTARY
1.1.1.244
-
RECOMMENDED NAME
GeneOntology No.
methanol dehydrogenase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
methanol + NAD+ = formaldehyde + NADH + H+
show the reaction diagram
ping-pong mechanism
-
methanol + NAD+ = formaldehyde + NADH + H+
show the reaction diagram
ping-pong mechanism in the absence of ACT protein
-
methanol + NAD+ = formaldehyde + NADH + H+
show the reaction diagram
ping-pong mechanism in the absence of ACT protein
Bacillus methanolicus C1
-
-
methanol + NAD+ = formaldehyde + NADH + H+
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Metabolic pathways
-
Methane metabolism
-
methanol oxidation to formaldehyde II
-
Microbial metabolism in diverse environments
-
SYSTEMATIC NAME
IUBMB Comments
methanol:NAD+ oxidoreductase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
dehydrogenase, methanol
-
-
-
-
EC 1.1.99.8
-
related
EC 1.1.99.8
Methylomicrobium sp.
-
related
EC 1.1.99.8
-
related
-
MDH
B3FR84
-
MDH
Methylomicrobium sp.
-
-
MDH
Methylophaga spp.
-
-
mxaF
Methylophaga spp.
-
-
NAD+ dependent methanol dehydrogenase
-
-
NAD+ dependent methanol dehydrogenase
-
-
-
XoxF
B3FR84
-
MEDH
-
-
-
-
additional information
-
not identical with methanol dehydrogenase which uses pyrroloquinoline quinone as prosthetic group, EC 1.1.99.8
additional information
Bacillus methanolicus C1
-
not identical with methanol dehydrogenase which uses pyrroloquinoline quinone as prosthetic group, EC 1.1.99.8
-
additional information
-
not identical with methanol dehydrogenase which uses pyrroloquinoline quinone as prosthetic group, EC 1.1.99.8
additional information
-
not identical with methanol dehydrogenase which uses pyrroloquinoline quinone as prosthetic group, EC 1.1.99.8
-
CAS REGISTRY NUMBER
COMMENTARY
74506-37-9
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
strains NRIC0498T, LMG1667 and NRIC0554
-
-
Manually annotated by BRENDA team
ATCC53907, three isozymes, two of the genes are encoded on the chromosome and one on a plasmid; NCIMB13113, three isozymes, two of the genes are encoded on the chromosome and one on a plasmid
-
-
Manually annotated by BRENDA team
thermotolerant methylotrophic strain C1
-
-
Manually annotated by BRENDA team
Bacillus methanolicus C1
strain C1
-
-
Manually annotated by BRENDA team
ATCC53907, three isozymes, two of the genes are encoded on the chromosome and one on a plasmid
-
-
Manually annotated by BRENDA team
NCIMB13113, three isozymes, two of the genes are encoded on the chromosome and one on a plasmid
-
-
Manually annotated by BRENDA team
strains B18LD and OH75-2a
UniProt
Manually annotated by BRENDA team
strain MP2, isolated from seawater, gene mxaF
-
-
Manually annotated by BRENDA team
Methylobacterium lusitanum MP2
strain MP2, isolated from seawater, gene mxaF
-
-
Manually annotated by BRENDA team
strain MP1, isolated from seawater, gene mxaF
-
-
Manually annotated by BRENDA team
Methylobacterium oryzae MP1
strain MP1, isolated from seawater, gene mxaF
-
-
Manually annotated by BRENDA team
sp. nov., isolated from seawater, gene mxaF
-
-
Manually annotated by BRENDA team
Methylomicrobium sp.
strain HG-1
-
-
Manually annotated by BRENDA team
Methylophaga spp.
-
-
-
Manually annotated by BRENDA team
no activity in Acetobacter lovaniensis
-
-
-
Manually annotated by BRENDA team
no activity in Acetobacter malorum
-
-
-
Manually annotated by BRENDA team
no activity in Acetobacter malorum DSM14337T
-
-
-
Manually annotated by BRENDA team
no activity in Acetobacter pomorum
-
-
-
Manually annotated by BRENDA team
no activity in Acetobacter pomorum DSM11825T
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
evolution
-
strain MGA3 contains three isozymes that belong to the type III Fe-NAD+-dependent alcohol dehydrogenases, but show a distinct substrate specificity and major differences with respect to transcriptional regulation of the paralogous genes; strain PB1 contains three isozymes that belong to the type III Fe-NAD+-dependent alcohol dehydrogenases, but show a distinct substrate specificity and major differences with respect to transcriptional regulation of the paralogous genes
evolution
-
strain MGA3 contains three isozymes that belong to the type III Fe-NAD+-dependent alcohol dehydrogenases, but show a distinct substrate specificity and major differences with respect to transcriptional regulation of the paralogous genes; strain PB1 contains three isozymes that belong to the type III Fe-NAD+-dependent alcohol dehydrogenases, but show a distinct substrate specificity and major differences with respect to transcriptional regulation of the paralogous genes
-
physiological function
-
biological significance of Mdh for methanol oxidation during methylotrophic growth and biological role of the enzyme as part of a formaldehyde detoxification system in the methanol consuming cells
physiological function
-
biological significance of Mdh for methanol oxidation during methylotrophic growth and biological role of the enzyme as part of a formaldehyde detoxification system in the methanol consuming cells
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1,3-propandiol + NAD+
? + NADH + H+
show the reaction diagram
-
very low activity, low activity
-
-
?
butanol + NAD+
butanal + NADH + H+
show the reaction diagram
-
-
-
-
?
ethanol + NAD+
ethanal + NADH
show the reaction diagram
-
-
-
-
-
ethanol + NAD+
ethanal + NADH
show the reaction diagram
-
best substrate
-
-
-
ethanol + NAD+
ethanal + NADH
show the reaction diagram
-
best substrate
-
-
r
ethanol + NAD+
acetaldehyde + NADH + H+
show the reaction diagram
-
-
-
-
r
formaldehyde + NADH + H+
methanol + NAD+
show the reaction diagram
-
Mdhs generally show higher activity and affinity for formaldehyde compared to methanol
-
-
r
isopropanol + NAD+
isopropanal + NADH + H+
show the reaction diagram
-
low activity
-
-
?
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
-
-
-
?
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
-
-
-
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
-
-
-
?
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
-
-
-
r
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
-
-
-
?
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
Methylomicrobium sp.
-
-
-
-
?
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
33% of activity with ethanol
-
r
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
44% of activity with ethanol, NADP+ cannot replace NAD+
-
-
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
no activity with 2-propanol, 2,3-butandiol, mannitol and glycerol
-
-
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
involved in initial methanol oxidation
-
r
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
involved in initial methanol oxidation
-
r
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
involved in initial methanol oxidation
-
r
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
involved in initial methanol oxidation
-
r
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
involved in the utilization of 1-carbon compounds as sole carbon source
-
-
?
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
Methylophaga spp.
-
screening of 1500 clones for the gene encoding the large subunit of MDH (mxaF) identifies a 25 kb insert containing fosmid (Fos-B11) with an mxaF amplicon of the correct size. Following shotgun sequencing of this insert, a ca. 9 kb contig is assembled that contains a cluster of sequences involved in the structure, assembly and regulation of MDH
-
-
?
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
strains grow on methanol
-
-
?
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
B3FR84, -
XoxF may not be involved in methylotrophy
-
-
?
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
very low activity, Mdhs generally show higher activity and affinity for formaldehyde compared to methanol, very low activity. Mdhs generally show higher activity and affinity for formaldehyde compared to methanol
-
-
r
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
-, involved in the utilization of 1-carbon compounds as sole carbon source
-
-
?
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
-
-
-
?
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
-, very low activity, Mdhs generally show higher activity and affinity for formaldehyde compared to methanol, very low activity. Mdhs generally show higher activity and affinity for formaldehyde compared to methanol
-
-
r
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
Bacillus methanolicus C1
-
-
-
-
?
n-butanol + NAD+
n-butanal + NADH
show the reaction diagram
-
87% of activity with ethanol
-
-
n-butanol + NAD+
n-butanal + NADH
show the reaction diagram
-
87% of activity with ethanol
-
r
n-propanol + NAD+
n-propanal + NADH
show the reaction diagram
-
71% of activity with ethanol
-
-
n-propanol + NAD+
n-propanal + NADH
show the reaction diagram
-
71% of activity with ethanol
-
r
octanol + NAD+
octanal + NADH
show the reaction diagram
Methylomicrobium sp., Methylomicrobium sp. HG-1
-
-
-
-
?
propanol + NAD+
propanal + NADH + H+
show the reaction diagram
-
-
-
-
?
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
-, very low activity, Mdhs generally show higher activity and affinity for formaldehyde compared to methanol, very low activity. Mdhs generally show higher activity and affinity for formaldehyde compared to methanol
-
-
r
additional information
?
-
-
all Mdh isozymes catalyze the oxidation of methanol, but the catalytic activity for methanol is considerably lower than for most other alcohols tested. The isozymes exhibit a broad and different substrate specificity range, and display both dehydrogenase and reductase activities. Isozyme Mdh3 shows the highest activity of all isozymes
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ethanol + NAD+
ethanal + NADH
show the reaction diagram
-
-
-
-
-
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
-
-
-
?
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
-
-
-
r
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
involved in initial methanol oxidation
-
r
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
involved in initial methanol oxidation
-
r
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
involved in initial methanol oxidation
-
r
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
involved in initial methanol oxidation
-
r
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
involved in the utilization of 1-carbon compounds as sole carbon source
-
-
?
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
-
-
-
r
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
Bacillus methanolicus C1
-
-
-
-
?
methanol + NAD+
formaldehyde + NADH + H+
show the reaction diagram
-
-
-
-
r
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
NAD+
-
tightly but not covalentely bound to enzyme
NAD+
-
tightly but not covalentely bound to enzyme, a second NAD+ is required for the regeneration of bound NADH
NAD+
Methylomicrobium sp.
-
-
NAD+
-
dependent on
additional information
-
no pyrroloquinoline quinone enzyme
-
additional information
-
enzyme contains most probably 1 NAD(H) cofactor molecule/subunit serving as primary electron acceptor, exogenous NAD+ (coenzyme) is responsible for reoxidation of enzyme bound NADH
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Mg2+
-
required, 1-2 mol per mol subunit, atomic absorption spectroscopy, activation by activator protein depends on Mg2+, Zn2+, Mn2+, Ca2+ cannot replace Mg2+
Mg2+
-
bound to enzyme, essential for binding of NAD+
Zn2+
-
required, 1 mol per mol subunit, atomic absorption spectroscopy
Zn2+
-
bound to enzyme
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1,10-phenanthroline
-
reduction of aldehyde
EDTA
-
reduction of aldehyde
iodoacetate
-
weak inhibition of aldehyde reduction
N-ethylmaleimide
-
weak inhibition of aldehyde reduction
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Activator protein
-
activation in presence of Mg2+, in vitro: 3 mol soluble activator protein per mol enzyme, in vivo estimated as 1 mol per 17.5 mol enzyme, MW 50000 with two subunits of MW 27000 each, in presence of activator protein and Mg2+ enzyme possesses a high affinity active site for alcohol and NAD+, up to 40fold increase of activity, primarily of Vmax, slight decrease of Km-value for methanol, enzyme/activator-interaction is dilution-sensitive, no stimulation of formaldehyde reduction
-
Activator protein
-
-
-
Activator protein
-
each activator protein subunit binds one molecule of NADH, activator may facilitate oxidation of reduced enzyme bound NADH cofactor
-
NudF protein
-
can be catalytically stimulated by the Bacillus subtilis NudF protein in vitro
-
activator protein ACT
-
all three isozymes are activated by ACT, which is encoded by gene act, overview
-
additional information
-
strongly stimulated by the ACT protein
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1
-
formaldehyde
-
pH 6.7, 45C, isozyme Mdh3
1.1
-
formaldehyde
-
pH 6.7, 45C, isozyme Mdh1
2
-
formaldehyde
-
-
3
-
formaldehyde
-
pH 6.7, 45C, isozyme Mdh1
4.5
-
formaldehyde
-
pH 6.7, 45C, isozyme Mdh2
7
-
formaldehyde
-
pH 6.7, 45C, isozyme Mdh2
0.02
-
NAD+
-
native enzyme, pH 9.5, 50C
0.04
-
NAD+
-
recombinant enzyme, pH 9.5, 50C
0.04
-
NAD+
-
recombinant enzyme without added Mg2+, pH 9.5, 50C
0.2
-
NAD+
-
S97T mutant enzyme without added Mg2+, pH 9.5, 50C
2.5
-
NAD+
-
S97G mutant enzyme without added Mg2+, pH 9.5, 50C
7.1
-
formaldehyde
-
pH 6.7, 45C, isozyme Mdh3
additional information
-
additional information
-
3.8 mM and 166 mM, methanol, due to two active sites biphasic kinetics for methanol, ethanol or NAD+ in cell-free extracts, not for formaldehyde
-
additional information
-
additional information
-
biphasic kinetics with all alcoholic substrates and NAD+ in the presence of activator protein
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
1.2
-
-
enzyme activity in crude extracts of methanol grown cells, fully activated enzyme, enzyme activity is 10fold lower in the absence of activator protein
19.6
-
-
formaldehyde reductase activity
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.7
-
-
formaldehyde reduction
6.7
-
-
formaldehyde reduction assay at
9
-
Methylomicrobium sp.
-
-
9.5
-
-
methanol oxidation
9.5
-
-
alcohol oxidation reaction assay at
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
45
-
-
activity optimum at about 45C for all isozymes
60
-
Methylomicrobium sp.
-
-
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.4
-
Methylomicrobium sp.
-
-
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Bacillus methanolicus C1
-
-
-
Manually annotated by BRENDA team
-
intracytoplasmic membrane
Manually annotated by BRENDA team
-
intracytoplasmic membrane
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
71000
-
-
SDS-PAGE and mass spectrometry
120000
-
Methylomicrobium sp.
-
gel filtration
363000
-
-
ultracentrifugation sedimentation equilibrium, dissociation occurs during the centrifugation time of 80 h
430000
-
-
calculated from electron microscopic analysis and MW of subunit
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
decamer
-
decameric structure detected by electron microscopic analysis
decamer
-
10 * 43000
decamer
-
10 * 43000, SDS-PAGE
decamer
Bacillus methanolicus C1
-
10 * 43000
-
dimer
-
alpha, beta, 1 * 10000 + 1 * 60000
dimer
Methylomicrobium sp.
-
2 * 60000, gel filtration
dimer
-
alpha, beta, 1 * 10000 + 1 * 60000
-
dimer
-
2 * 60000, gel filtration
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
45
-
-
20 min, the isozymes are all stable up to; 20 min, the isozymes are stable up to, except for isozyme Mdh1, which shows about 35-40% loss of activity
50
-
-
t1/2: above 2 min
60
-
-
t1/2: 6 min
60
-
Methylomicrobium sp.
-
remains stable at 60C for 20 min
60
-
-
20 min, isozyme Mdh1 is inactivated, isozyme Mdh2 loses about 80% of activity, while isozyme Mdh3 retains over 80% of its activity; 20 min, isozyme Mdh1 loses 90% activity, isozyme Mdh2 loses 80% of activity, while isozyme Mdh3 retains 60% of its activity
70
-
-
t1/2: 2 min
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
dilution inactivates methanol dehydrogenase activity, not reductase activity, sucrose, 20% w/v, stabilizes formaldehyde reductase not methanol dehydrogenase activity, metal ions, such as Fe2+ and Zn2+, do not stabilize methanol dehydrogenase activity, glycerol does not stabilize methanol dehydrogenase activity, dithiothreitol stabilizes reductase, not methanol dehydrogenase activity, storage leads to dissociation
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-80 to 4C, in the presence of DTT at least 4 days stable
-
4-20C, complete dissociation after 48 h
-
4-20C, enzyme dissociates upon storage
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
native and recombinant proteins
-
purification of activator protein
-
Q-Sepharose, Phenyl-Superose, dehydrogenase activity is almost completely lost after purification, 34% reductase activity are recovered
-
wild type and mutant enzymes
-
gel filtration
-
-
Methylomicrobium sp.
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Escherichia coli DH5alpha
-
expressed in Escherichia coli, full activity only when grown in Mg2+ containing medium
-
three isozymes, DNA and amino acid sequence determination and analysis, expression in Escherichia coli strain ER2566
-
gene mxaF, DNA and amino acid sequence determination and analysis, genotypic and phylogenetic analysis and species verfication, overview
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D100N
-
strongly reduced NAD+-binding, no activity
D88N
-
only minor effects on activity
G13A
-
only minor effects on activity
G15A
-
only minor effects on activity
G95A
-
impaired cofactor binding, low acitivity
K103R
-
strongly reduced NAD+-binding, no activity
S97G
-
impaired cofactor binding, much higher acitivity than the wild type enzyme, no activation with ACT protein
S97T
-
impaired cofactor binding, much higher acitivity than the wild type enzyme, no activation with ACT protein