Information on EC 1.14.13.25 - methane monooxygenase (soluble)

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

EC NUMBER
COMMENTARY hide
1.14.13.25
-
RECOMMENDED NAME
GeneOntology No.
methane monooxygenase (soluble)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
methane + NAD(P)H + H+ + O2 = methanol + NAD(P)+ + H2O
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Metabolic pathways
-
-
Methane metabolism
-
-
methane oxidation to methanol I
-
-
Microbial metabolism in diverse environments
-
-
nitrite-dependent anaerobic methane oxidation
-
-
SYSTEMATIC NAME
IUBMB Comments
methane,NAD(P)H:oxygen oxidoreductase (hydroxylating)
The enzyme is soluble, in contrast to the particulate enzyme, EC 1.14.18.3. Broad specificity; many alkanes can be hydroxylated, and alkenes are converted into the corresponding epoxides; CO is oxidized to CO2, ammonia is oxidized to hydroxylamine, and some aromatic compounds and cyclic alkanes can also be hydroxylated, but more slowly.
CAS REGISTRY NUMBER
COMMENTARY hide
51961-97-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
protein A fragment, gene pmoA; a gammaproteobacterium closely related to methanotrophs
SwissProt
Manually annotated by BRENDA team
; HD6T
-
-
Manually annotated by BRENDA team
Methylocystis sp.
Methylocystis sp. WI 14
strain WI 14
-
-
Manually annotated by BRENDA team
Methyloferula stellata gen. nov., sp. nov.
gene mmoX
-
-
Manually annotated by BRENDA team
alpha-subunit, strain LAY; isolated from acidic forest soil near Marburg, Germany, gene mmoX
UniProt
Manually annotated by BRENDA team
alpha-subunit, strain SOP9; isolated from an acidic peat soil sampled at a depth of 10 to 20 cm of the Sphagnum peat bog Bakchar, West Siberia, Russia, gene mmoX
UniProt
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
strain 5
-
-
Manually annotated by BRENDA team
strain 5
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-
Manually annotated by BRENDA team
strain IMV 3011
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-
Manually annotated by BRENDA team
methylotrophic bacterium
-
-
-
Manually annotated by BRENDA team
alpha-subunit; a type I methanotroph
UniProt
Manually annotated by BRENDA team
alpha-subunit; a type I methanotroph
UniProt
Manually annotated by BRENDA team
synthetic construct
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1,1,2,2-tetramethylcyclopropane + NADH + O2
?
show the reaction diagram
-
-
-
-
?
1-butene + NAD(P)H + O2
1,2-epoxybutane + NAD(P)+ + H2O
show the reaction diagram
2,3-dimethylpentane + NAD(P)H + O2
3,4-dimethylpentan-2-ol + NAD(P)+ + H2O
show the reaction diagram
-
-
-
?
2-methylpropane + NAD(P)H + O2
2-methylpropan-2-ol + 2-methylpropan-1-ol + NAD(P)+ + H2O
show the reaction diagram
-
-
-
?
adamantane + NAD(P)H + O2
1-adamantanol + 2-adamantanol + NAD(P)+ + H2O
show the reaction diagram
-
-
-
?
ammonia + NAD(P)H + O2
hydroxylamine + NAD(P)+ + H2O
show the reaction diagram
-
-
-
-
?
ammonia + NADH + H+ + O2
?
show the reaction diagram
-
-
-
-
?
benzene + NAD(P)H + O2
cyclohexanol + phenol + hydroquinone + NAD(P)+ + H2O
show the reaction diagram
benzene + NAD(P)H + O2
phenol + NAD(P)+ + H2O
show the reaction diagram
benzene + NADH + H+ + O2
?
show the reaction diagram
-
-
-
-
?
beta-pinene + NAD(P)H + O2
6,6-dimethylbicyclo[3.1.1]hept-2-ene-2-methanol + beta-pinene oxide + NAD(P)+ + H2O
show the reaction diagram
-
-
-
?
biphenyl + NAD(P)H + H+ + O2
2-hydroxybiphenyl + 4-hydroxybiphenyl + NAD(P)+ + H2O
show the reaction diagram
-
-
-
-
?
bromobenzene + NAD(P)H + O2
bromophenol + NAD(P)+ + H2O
show the reaction diagram
Methylocystis sp.
-
sMMO
-
?
bromomethane + NAD(P)H + O2
?
show the reaction diagram
bromomethane + NADH + H+ + O2
?
show the reaction diagram
-
-
-
-
?
butane + NAD(P)H + O2
1-butanol + 2-butanol + NAD(P)+ + H2O
show the reaction diagram
butylene + NAD(P)H + O2
butylene oxide + NAD(P)+ + H2O
show the reaction diagram
Methylocystis sp.
-
sMMO
-
?
carbon monoxide + NADH + H+ + O2
?
show the reaction diagram
-
-
-
-
?
chlorobenzene + NAD(P)H + O2
chlorophenol + NAD(P)+ + H2O
show the reaction diagram
Methylocystis sp.
-
sMMO
-
?
chloromethane + NAD(P)H + O2
formaldehyde + NAD(P)+ + H2O + ?
show the reaction diagram
-
-
-
?
chloromethane + NADH + H+ + O2
?
show the reaction diagram
-
-
-
-
?
chloronaphthalene + NAD(P)H + O2
chloronaphthol + NAD(P)+ + H2O
show the reaction diagram
Methylocystis sp.
-
sMMO
-
?
chloropentane + NAD(P)H + O2
chloropentanol + NAD(P)+ + H2O
show the reaction diagram
Methylocystis sp.
-
sMMO
-
?
cis-1,3-dimethylcyclohexane + NAD(P)H + O2
3,5-dimethylcyclohexanol + 1-cis-3-dimethylcyclohexanol + NAD(P)+ + H2O + 1-trans-3-dimethylcyclohexanol
show the reaction diagram
-
-
1-trans-3-dimethylcyclohexanol is produced in a low concentration
?
cis-1,4-dimethylcyclohexane + NAD(P)H + O2
1-cis-4-dimethylcyclohexanol + NAD(P)+ + H2O + trans-2,5-dimethylcyclohexanol
show the reaction diagram
-
-
trans-2,5-dimethylcyclohexanol is produced in a low concentration
?
cis-2-butene + NAD(P)H + O2
cis-2,3-epoxybutane + cis-2-buten-1-ol + 2-butanone + NAD(P)+ + H2O
show the reaction diagram
CO + NAD(P)H + O2
CO2 + NAD(P)+ + H2O
show the reaction diagram
cyclohexane + NAD(P)H + O2
cyclohexanol + NAD(P)+ + H2O
show the reaction diagram
cyclohexene + NAD(P)H + O2
epoxycyclohexane + 2-cyclohexen-1-ol + NAD(P)+ + H2O
show the reaction diagram
-
-
-
?
cytochrome c + NAD(P)H + O2
reduced cytochrome c + NAD(P)+ + H2O
show the reaction diagram
Methylocystis sp.
-
sMMO
-
-
?
dichloromethane + NAD(P)H + O2
CO + Cl- + NAD(P)+ + H2O
show the reaction diagram
-
-
-
?
diethyl ether + NAD(P)H + O2
ethanol + ethanal + NAD(P)+ + H2O
show the reaction diagram
difluoromethane + NADH + O2
difluoromethanol + NAD+ + H2O
show the reaction diagram
dimethyl ether + NAD(P)H + O2
methanol + formaldehyde + NAD(P)+ + H2O
show the reaction diagram
dimethyl ether + NADH + H+ + O2
?
show the reaction diagram
-
-
-
-
?
ethane + NAD(P)H + O2
ethanol + NAD(P)+ + H2O
show the reaction diagram
ethane + NADH + O2
?
show the reaction diagram
-
-
-
-
?
ethane + NADH + O2
ethanol + NAD+ + H2O
show the reaction diagram
-
-
-
-
?
ethene + NAD(P)H + O2
epoxyethane + NAD(P)+ + H2O
show the reaction diagram
ethylbenzene + NAD(P)H + H+ + O2
1-phenylethanol + 3-ethylphenol + 4-ethylphenol + NAD(P)+ + H2O
show the reaction diagram
-
-
-
-
?
fluorobenzene + NAD(P)H + O2
fluorophenol + NAD(P)+ + H2O
show the reaction diagram
Methylocystis sp.
-
sMMO
-
?
fluoromethane + NADH + O2
fluoromethanol + NAD+ + H2O
show the reaction diagram
formate + NAD(P)H + O2
?
show the reaction diagram
-
assay with whole cells
-
-
?
furan + NAD(P)H + O2
?
show the reaction diagram
-
-
-
-
?
furan + NADH + O2
? + NAD+ + H2O
show the reaction diagram
-
-
-
-
?
heptane + NAD(P)H + O2
1-heptanol + 2-heptanol + NAD(P)+ + H2O
show the reaction diagram
hexane + NAD(P)H + O2
1-hexanol + 2-hexanol + NAD(P)+ + H2O
show the reaction diagram
isobutane + NAD(P)H + O2
2-methyl-1-propanol + 2-methyl-2-propanol + NADP+ + H2O
show the reaction diagram
-
-
-
?
isopentane + NAD(P)H + O2
2-methylbutan-1-ol + 3-methylbutan-1-ol + 2-methylbutan-2-ol + 3-methylbutan-2-ol + NADP+ + H2O
show the reaction diagram
-
-
-
?
methane + duroquinol + O2
methanol + duroquinone + H2O
show the reaction diagram
methane + NAD(P)H + H+ + O2
methanol + NAD(P)+ + H2O
show the reaction diagram
-
-
-
-
?
methane + NAD(P)H + O2
methanol + NAD(P)+ + H2O
show the reaction diagram
methane + NADH + H+ + O2
methanol + H2O + NAD+
show the reaction diagram
-
-
-
-
?
methane + NADH + H+ + O2
methanol + NAD+ + H2O
show the reaction diagram
methane + NADH + O2
methanol + NAD+ + H2O
show the reaction diagram
methane + trans-dichloroethylene + vinyl chloride + trichloroethylene + ?
formaldehyde + ?
show the reaction diagram
-
each of these compounds is completely degraded by sMMO-expressing cells when initial concentrations are either 0.01 or 0.03 mM
-
-
?
methanol + NADH + H+ + O2
? + H2O + NAD+
show the reaction diagram
-
substrate of intermediate species, Hperoxo and Q, kinetics, overview
-
-
?
methylamine + NADH + H+ + O2
hydroxymethylamine + H2O + NAD+
show the reaction diagram
-
substrate of intermediate species, Hperoxo and Q, kinetics, overview
-
-
?
methylcyanide + NADH + H+ + O2
hydroxymethylcyanide + H2O + NAD+
show the reaction diagram
-
substrate of intermediate species, Hperoxo and Q, kinetics, and proposed mechanism of CH3CN hydroxylation by Hperoxo, overview
-
-
?
methylene cyclohexane + NAD(P)H + O2
1-cyclohexane-1-methanol + methylene cyclohexane oxide + 4-hydroxymethylene cyclohexane + NAD(P)+ + H2O
show the reaction diagram
-
-
-
?
naphthalene + NAD(P)H + H+ + O2
alpha-naphthol + beta-naphthol + NAD(P)+ + H2O
show the reaction diagram
-
-
-
-
?
naphthalene + NAD(P)H + O2
alpha-naphthol + beta-naphthol + NAD(P)+ + H2O
show the reaction diagram
naphthalene + NADH + H+
alpha-naphthol + beta-naphthol + NAD+ + H2O
show the reaction diagram
nitrobenzene + NADH + O2
nitrophenol + NAD+ + H2O
show the reaction diagram
nitromethane + NADH + H+ + O2
?
show the reaction diagram
-
-
-
-
?
octane + NAD(P)H + O2
1-octanol + 2-octanol + NAD(P)+ + H2O
show the reaction diagram
-
-
-
?
pentane + NAD(P)H + O2
1-pentanol + 2-pentanol + NAD(P)+ + H2O
show the reaction diagram
phenylalanine + NAD(P)H + O2
tyrosine + NAD(P)+ + H2O
show the reaction diagram
-
-
-
?
propane + NAD(P)H + O2
1-propanol + 2-propanol + NAD(P)+ + H2O
show the reaction diagram
propene + NAD(P)H + O2
1,2-epoxypropane + NAD(P)+ + H2O
show the reaction diagram
propene + NADH + H+ + O2
epoxypropane + NAD+ + H2O
show the reaction diagram
propylaldehyde + NADH + H+ + O2
? + H2O + NAD+
show the reaction diagram
-
substrate of intermediate species, Hperoxo and Q, kinetics, overview
-
-
?
propylene + duroquinol + O2
propylene oxide + reduced duroquinol + H2O
show the reaction diagram
-
-
-
-
?
propylene + NAD(P)H + O2
propylene oxide + NADP+ + H2O
show the reaction diagram
propylene + NADH + H+ + O2
propylene epoxide + NAD+ + H2O
show the reaction diagram
-
-
-
-
?
propylene + NADH + O2
propylene + NADH + O2
show the reaction diagram
-
-
-
-
?
propylene + NADH + O2
propylene epoxide + NAD+ + H2O
show the reaction diagram
propylene + NADH + O2
propylene oxide + NAD+ + H2O
show the reaction diagram
pyridine + NAD(P)H + O2
pyridine N-oxide + NAD(P)+ + H2O
show the reaction diagram
-
-
-
?
pyridine + NADH + H+ + O2
?
show the reaction diagram
-
-
-
-
?
styrene + NAD(P)H + O2
styrene epoxide + NAD(P)+ + H2O
show the reaction diagram
styrene + NADH + H+ + O2
?
show the reaction diagram
-
-
-
-
?
toluene + NAD(P)H + H+ + O2
benzyl alcohol + cresol + NAD(P)+ + H2O
show the reaction diagram
-
-
-
-
?
toluene + NAD(P)H + O2
benzyl alcohol + cresol + NAD(P)+ + H2O
show the reaction diagram
-
-
-
?
toluene + NAD(P)H + O2
benzyl alcohol + NAD(P)+ + H2O
show the reaction diagram
-
-
-
?
toluene + NAD(P)H + O2
cresol + NAD(P)+ + H2O
show the reaction diagram
Methylocystis sp.
-
sMMO
-
?
trans-2-butene + NAD(P)H + O2
trans-2,3-epoxybutane + trans-2-buten-1-ol + NAD(P)+ + H2O
show the reaction diagram
trichloromethane + NAD(P)H + O2
CO2 + Cl- + NAD(P)+ + H2O
show the reaction diagram
-
-
-
?
trichloromethane + NADH + H+ + O2
?
show the reaction diagram
-
-
-
-
?
xylene + NAD(P)H + O2
xylenol + NAD(P)+ + H2O
show the reaction diagram
Methylocystis sp.
-
sMMO
-
?
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
formate + NAD(P)H + O2
?
show the reaction diagram
-
assay with whole cells
-
-
?
methane + duroquinol + O2
methanol + duroquinone + H2O
show the reaction diagram
methane + NAD(P)H + O2
methanol + NAD(P)+ + H2O
show the reaction diagram
methane + NADH + H+ + O2
methanol + H2O + NAD+
show the reaction diagram
-
-
-
-
?
methane + NADH + H+ + O2
methanol + NAD+ + H2O
show the reaction diagram
methane + NADH + O2
methanol + NAD+ + H2O
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cytochrome c
-
one of three protein components is a soluble CO-binding cytochrome c
Ferredoxin
-
several MMOR ferredoxin analogues, intermolecular electron transfer from ferredoxin analogues to hydroxylase protein MMOH, redox potential determinations, overview
-
additional information
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
copper
-
the enzyme expresses the soluble enzyme form under copper limitation, and the membrane-bound particulate MMO at high copper-to-biomass ratio, mechanism of the copper switch involves a tetrameric 480 kDA sensor protein MmoS, encoded by gene mmoS, as part of a two-component signaling system, domain organization, MmoS contains a FAD cofactor, indirect regulation without binding of copper to MmoS, overview
Ni2+
-
protein B contains 0.04 mol Ni2+ per mol protein
[2Fe-2S] cluster
-
bound to the MMOR enzyme component
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,3-Dimercaptopropan-1-ol
-
-
2,4-Dichloro-(6-phenylphenoxy)ethylamine hydrochloride
2,4-Dichloro-(6-phenylphenoxy)ethyldiethylamine
-
-
2-mercaptoethanol
-
-
3-Amino-1,2,4-triazole
-
-
8-hydroxyquinoline
Acetylene
Allylthiourea
ammonium chloride
-
slightly
Cd2+
-
soluble enzyme form more than the membrane-bound form
Chloramphenicol
-
sMMO
Co2+
Methylocystis sp.
-
slightly, sMMO
Dichloromethane
-
competitively in presence of formate
Dimercaptopropanol
-
-
Dithionite
-
34% inhibition at 5 mM
dithiothreitol
-
-
DTT
-
24% inhibition at 20 mM
ethyl carbamate
-
-
glycerol
-
40% inhibition at 15%
Hg2+
Methylocystis sp.
-
complete inhibition at 0.01 mM
methylamine
-
slightly
N2
-
N2 atmosphere causes 28% inhibition
o-phenanthroline
-
-
poly-beta-hydroxybutyrate
protein MMOB
-
inhibition of sMMO auxiliary proteins MMOB or MMOD severely diminishes electron-transfer throughput from MMOR, primarily by shifting the bulk of electron transfer to the slowest pathway, the biphasic reactions for electron transfer to MMOH from several MMOR ferredoxin analogues are also inhibited, overview
-
protein MMOD
-
inhibition of sMMO auxiliary proteins MMOB or MMOD severely diminishes electron-transfer throughput from MMOR, primarily by shifting the bulk of electron transfer to the slowest pathway, the biphasic reactions for electron transfer to MMOH from several MMOR ferredoxin analogues are also inhibited, overview
-
Thiosemicarbazide
-
-
trichlorethylene
-
non-competitively; pMMO
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
catalase
-
reduction current is enhanced by the presence of catalase ot if the reaction is performed in a flow-cell, probably because O2 is reduced to H2O2, by the hydroxylase component of the enzyme MMOH at the electrode surface and the H2O2 then inactivates the enzyme unless removed by catalase or a continous flow solution
-
regulatory protein MMOB
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the native regulatory protein MMOB is required for maximum enzyme activity
-
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.003 - 0.066
methane
0.0064 - 0.0558
NADH
5.2
NADPH
Methylocystis sp.
-
sMMO component C reductase
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
7.6
furan
Methylosinus trichosporium
-
-
3.7
methane
Methylosinus trichosporium
-
-
4.4
Propene
Methylosinus trichosporium
-
-
0.23
propylene
Methylococcus capsulatus
-
25C, purified enzyme
additional information
additional information
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00011
-
purified enzyme, using propylene and duroquinol as substrates
0.0008
Methylocystis sp.
-
sMMO, substrate chloronaphthalene
0.0012
Methylocystis sp.
-
sMMO, substrate naphthalene
0.0029
-
membrane-bound enzyme, using propylene and NADH as substrates
0.003
-
membrane-bound enzyme, using propylene and duroquinol as substrates
0.0039
Methylocystis sp.
-
sMMO, substrate chloropentane
0.0122
Methylocystis sp.
-
sMMO, low copper growth concentration, growth substrate nitrate
0.0191
Methylocystis sp.
-
sMMO, substrate butylene
0.02 - 0.04
-
purified MMOH, substrate propylene
0.0254
Methylocystis sp.
-
sMMO, substrate propylene
0.0336
Methylocystis sp.
-
sMMO, substrate ethylene
0.072
-
component A
0.1
-
about, all 3 components individually
0.185
-
component A
0.205
Methylocystis sp.
-
sMMO, substrate diethylic ether
0.208
-
purified component A hydroxylase
0.265
-
protein A after reconstitution of iron
0.3 - 0.45
-
pH 7.0, 45C
0.334
Methylocystis sp.
-
sMMO, substrate propane
0.604
-
purified enzyme, substrate propylene
0.764
-
all components, substrate propene
1.66
-
sMMO protein B triple mutant G10A/G13Q/G16A
2.284
-
His-tagged sMMO protein B triple mutant G10A/G13Q/G16A
3.99
-
sMMO protein B mutant G13Q
5.09
-
sMMO, wild-type enzyme
5.71
-
purified component C
11.37
-
purified protein B
17.5
-
component C
26.1
-
purified enzyme, substrate propene
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 7.6
-
assay at
7.2
Methylocystis sp.
-
assay at
7.5
-
furan, propene
8.5 - 9
-
component C
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.4 - 7.4
-
pH 6.4: about 20% of activity maximum, pH 7.4: about 25% of activity maximum
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
assay at, hydroxylase component
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.2
-
isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
Methylococcus capsulatus (strain ATCC 33009 / NCIMB 11132 / Bath)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
9400
-
component: small protein, native PAGE
13000
-
component: CO-binding cytochrome c, native PAGE
15100
-
component B, gel filtration
15800
-
component B containing FAD and [Fe2-S2]-cluster, gel filtration
15850
-
component protein B of sMMO, mass spectrometry
17000
-
component: protein B, gel filtration
18000
Methylocystis sp.
-
component protein B of sMMO
24000
-
component D of sMMO, forms homodimers, gel filtration
32000
Methylocystis sp.
-
component B
38000 - 38550
-
component: protein C, gel filtration
38300 - 38400
-
component C: reductase, gel filtration
39700
-
component C NADH-reductase, gel filtration
40000
-
component C acceptor reductase
41000
Methylocystis sp.
-
component C reductase of sMMO
42000
-
component C
47000
-
component: copper-containing protein, native PAGE
201300
-
gel filtration
210000
-
component A, analytical ultracentrifugation
220000
229000
Methylocystis sp.
-
component A hydroxylase of sMMO
240000
-
component A hydroxylase
241000 - 246000
-
protein A hydroxylase, gel filtration
245000
-
protein A hydroxylase, gel filtration
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterotrimer
hexamer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure analysis
-
sitting drop vapor diffusion method, using 0.1 M MES (pH 6.5) and 15% PEG 20000 (w/v)
-
X-ray crystal structure of the apo, Mn(II)-soaked, and Co(II)-grown hydroxylase component of methane monooxygenase, determined at 2.3 A or greater resolution, reveal that the presence of metal ions is essential for the proper folding of helices E, F, and H of the alpha-subunit
-
sitting drop vapour diffusion method with 100 mM cacodylate, pH 6.5, 20% (v/v) PEG 3000, 250 mM magnesium formate or MnCl2
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 7
-
protein B, stable, rapid loss of activity above and below
438930
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45
-
protein B, half-life: 10.2 min
55
-
protein A, 10 min
additional information
Methylocystis sp.
-
N-terminal truncated sMMO component B increases the heat stability of the sMMO hydroxylase
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
5 mM thioglycollate, 5 mM dithiothreitol, 5 mM NADH stabilize component C at 0C
-
Fe2+ stabilizes component C of sMMO
Methylocystis sp.
-
instability of enzyme in crude extract
instability of the enzyme during purification is probably due to loss of iron
-
no component of the enzyme is stable to freezing
-
protein A unstable to successive freezing and thawing
-
protein B stable to successive freezing and thawing
-
protein B, in crude form requires addition of protease inhibitor phenylmethylsulfonyl fluoride
-
protein C requires presence of thiol protective agent, e.g. sodium thioglycolate throughout purification
-
sMMO is more stable and easier to purify than pMMO
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 50% glycerol, component A, stable for several weeks
-
-80C, more than 1 year
-
0C, protein C, loss of 60-90% of activity within 20 h
-
0C, proteins A and B, stable for at least 24 h
-
4C, sMMO component C reductase, 90% loss of activity after 120 h, can be restored by Fe2+
Methylocystis sp.
-
sMMO component A hydroxylase is very unstable, splits into inactive subunits when stored frozen even for short periods
Methylocystis sp.
-
sMMO component protein B is stable when stored frozen
Methylocystis sp.
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
3 components: a soluble CO-binding cytochrome c, a copper-containing protein, another small protein
-
component B of sMMO
Methylocystis sp.
-
component D of sMMO recombinant from Escherichia coli as His-tagged thioredoxin-fusion protein, the thioredoxin is cleaved off during purification by factor Xa
-
DEAE-Sepharose column chromatography, Q sepharose column chromatography, S-200 gel filtration, and S-300 gel filtration
-
hydroxylase component
-
hydroxylase component A
-
improved purification protocol using stabilizing agents
-
native enzyme 8.08fold by anion exchange chromatography, ultrafiltration, gel filtration, and again anion exchange chromatography, to 95% purity
-
native MMOH, in the purification procedure includes anion exchange chromatography and gel filtration
-
protein A and C
-
recombinant component protein B of sMMO as glutathione-S-transferase fusion protein from Escherichia coli
-
recombinant enzyme component MMOB from Escherichia coli by solubilization from the 12000 x g pellet with 8 M urea, followed by dilution for enzyme refolding, and gel filtration
-
sMMO protein B wild-type and mutants recombinant from Escherichia coli by affinity chromatography, high salt concentration increases the binding stability between protein B and hydroxylase of sMMO
-
sMMO with all components
Methylocystis sp.
-
Source 15Q column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, phylogenetic analysis, quantitative expression analysis
DNA and amino acid sequence determination and analysis. The sMMO gene cluster consists of the structural genes mmoXYBZDC, the regulatory genes mmoG and mmoR and another ORF orf1. These sMMO genes are transcribed as a single unit from a s54-dependent promoter located upstream of mmoX
DNA sequence analysis; overexpression of an additional protein component D of sMMO encoded by orfY as thioredoxin-fusion protein with His-Tag in Escherichia coli, protein component D is termed MMOD
-
expressed in Escherichia coli
expression in Escherichia coli
-
expression in Escherichia coli, determination of complete sMMO DNA gene sequence, phylogenetic analysis
Methylocystis sp.
-
expression of component protein B of sMMO as glutathione-S-transferase fusion protein in Escherichia coli
-
expression of His-tagged triple mutant G10A/G13Q/G16A and G13Q mutant in Escherichia coli
-
gene mmoX, DNA and amino acid sequence determination and analysis; gene mmoX, DNA and amino acid sequence determination and analysis; gene mmoX, DNA and amino acid sequence determination and analysis
mutants are expressed in sMMO-negative cells of Methylosinus trichosporium
-
overexpression of enzyme component MMOB in Escherichia coli
-
phylogenetic analysis
-
transcription of sMMO is repressed at Cu2+ concentration above 0.00086 mM per g dry cell weight
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
activity of pMMO in whole cells is approximately 10fold greater than that of the purified sample
-
activity of pMMO in whole cells is approximately 10fold greater than that of the purified sample; the sMMO enzyme is expressed when cells are essentially starved for copper and the copper-to-biomass ratio is low (less than 0.0002 mM Cu2+)
-
-
necessary for the expression are a sigma(54)-dependent transcriptional activator, MmoR, and a putative GroEL-like chaperone, MmoG
-
sMMO is expressed at low copper/biomass ratios
sMMO is produced when the copper/biomass ratio is low
-
the sMMO enzyme is expressed when cells are essentially starved for copper and the copper-to-biomass ratio is low (less than 0.0002 mM Cu2+)
transcription of sMMO is arrested in cells exposed to high levels of copper ions and sMMO mRNA transcripts are not detected 15 min after the addition of CuSO4
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
G10A/G13Q/G16A
-
His-tagged protein B of sMMO, triple mutant is resistant to degradation in contrast to the wild-type, N-terminus is responsible for unusual mobility in size exclusion chromatography and proteolytic sensitivity of protein B; reduced activity
G10A/G13Q/G16A
-
His-tagged protein B of sMMO, triple mutant is resistant to degradation in contrast to the wild-type, N-terminus is responsible for unusual mobility in size exclusion chromatography and proteolytic sensitivity of protein B; reduced activity