Information on EC 4.5.1.3 - dichloromethane dehalogenase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
4.5.1.3
-
RECOMMENDED NAME
GeneOntology No.
dichloromethane dehalogenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
dichloromethane + H2O = formaldehyde + 2 chloride
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
carbon-halide lyase reaction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Chloroalkane and chloroalkene degradation
-
-
Microbial metabolism in diverse environments
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-
SYSTEMATIC NAME
IUBMB Comments
dichloromethane chloride-lyase (adding H2O; chloride-hydrolysing; formaldehyde-forming)
Requires glutathione.
CAS REGISTRY NUMBER
COMMENTARY hide
97002-70-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain WZ-12 CCTCC M 207006
TrEMBL
Manually annotated by BRENDA team
DSM 1869
-
-
Manually annotated by BRENDA team
GJ21
Uniprot
Manually annotated by BRENDA team
KDM2
Uniprot
Manually annotated by BRENDA team
KDM4
Uniprot
Manually annotated by BRENDA team
DM11
-
-
Manually annotated by BRENDA team
strain TM1
-
-
Manually annotated by BRENDA team
strain TM1
-
-
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
1,2-dichloroethane + H2O
? + Cl-
show the reaction diagram
-
-
-
-
?
bromoethane + H2O
ethanal + Br-
show the reaction diagram
-
-
-
-
?
CH2BrCl + H2O
formaldehyde + Br- + Cl-
show the reaction diagram
CH2ClF + H2O
formaldehyde + Cl- + F-
show the reaction diagram
CH2ClI + H2O
formaldehyde + Cl- + I-
show the reaction diagram
chloroethane + H2O
ethanal + Cl-
show the reaction diagram
-
-
-
-
?
dibromomethane + H2O
formaldehyde + Br-
show the reaction diagram
dichlormethane + H2O
formaldehyde + chloride
show the reaction diagram
dichloromethane + H2O
formaldehyde + 2 Cl-
show the reaction diagram
dichloromethane + H2O
formaldehyde + chloride
show the reaction diagram
dichloromethane + H2O
formaldehyde + Cl-
show the reaction diagram
diiodomethane + H2O
formaldehyde + I-
show the reaction diagram
iodoethane + H2O
ethanal + I-
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
dichlormethane + H2O
formaldehyde + chloride
show the reaction diagram
dichloromethane + H2O
formaldehyde + 2 Cl-
show the reaction diagram
dichloromethane + H2O
formaldehyde + chloride
show the reaction diagram
dichloromethane + H2O
formaldehyde + Cl-
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
glutathione
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(Z)-1,2-Dichloroethylene
-
-
1,1,2-trichloroethane
-
weak
1,1-Dichloroethane
-
-
1,2-Dichloroethane
-
-
1,3-Dichloropropane
-
-
1,4-Dichlorobutane
-
-
1-Chlorobutane
-
-
1-Chloropentane
-
-
1-Chloropropane
-
-
2,2-Dichloroethyl methyl ether
-
-
2-Chloroethyl methyl ether
-
-
chlorobenzene
-
-
Dichloroacetic acid
-
-
Dichloromethane
-
an inhibitory effect with increasing dichloromethane concentrations is observed at concentrations higher than 400 mg/l
Methyl dichloroacetate
-
-
Sodium azide
[2-(3-chlorophenyl)hydrazinylidene]propanedinitrile
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
glutathione
nigericin
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.33
Bromoethane
-
25°C, pH 8
0.00162 - 0.43
CH2BrCl
0.053
CH2ClF
-
25°C, pH 8
0.002 - 0.18
CH2ClI
0.33
chloroethane
-
25°C, pH 8
0.0005 - 0.338
Dibromomethane
0.009 - 0.47
Dichloromethane
0.0017 - 0.521
Diiodomethane
0.039
Iodoethane
-
25°C, pH 8
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.14
Bromomethane
Methylophilus sp.
-
25°C, pH 8
1
CH2BrCl
Methylophilus sp.
-
25°C, pH 8
0.009
CH2ClF
Methylophilus sp.
-
25°C, pH 8
1.03
CH2ClI
Methylophilus sp.
-
25°C, pH 8
0.1
chloroethane
Methylophilus sp.
-
25°C, pH 8
3 - 5.6
Dibromomethane
0.55 - 4.78
Dichloromethane
1.31
Diiodomethane
Methylophilus sp.
-
25°C, pH 8
0.15
Iodoethane
Methylophilus sp.
-
25°C, pH 8
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.112
-
W117Y with dichloromethane as substrate
0.436
-
W117Y with diiodomomethane as substrate
0.804
-
W117Y with CH2ClI as substrate
0.914
-
W117Y with CH2BrCl as substrate
0.98
-
W117Y with dibromomethane as substrate
1.041
-
hydroxylapatite eluate
3.336
-
wild-type with CH2BrCl as substrate
3.618
-
wild-type with CH2ClI as substrate
4.668
-
wild-type with dichloromethane as substrate
4.728
-
wild-type with diiodomomethane as substrate
7.398
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wild-type with dibromomethane as substrate
14.36
-
crude extract
21.38
-
after purification by (NH4)2SO4 salting out
49.12
-
after purification by Sephadex G-75 chromatography
118.8
-
after purification by anion exchange column
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
assay at; assay at; assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
assay at; assay at; assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
expression of gene dcmA; expression of gene METDI2657; expression of gene METDI2658
Manually annotated by BRENDA team
expression of gene dcmA; expression of gene METDI2657; expression of gene METDI2658
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20000
-
estimated by SDS-PAGE
34000
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x * 34000, SDS-PAGE
35000
-
x * 35000, SDS-PAGE
195000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
-
alpha6, 6 * 33000, SDS-PAGE
monomer
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 7.5
the enzyme retains more than 50% activity between pH 6.0 and 7.5 but the activity drops off markedly at a pH value above 7.5 or below 6.0
702713
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 40
more than 50% of the activity is observed between 25°C and 40°C
22
-
6 days, 50% loss of activity
45
-
after 5 min remains 38% activity of wild-type enzyme and 54% activity of W117Y
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, protein concentration 4 mg/ml, 100 mM potassium phosphate, pH 7.5, 2.5 mM DTT, 0.1 mM EDTA, 25% v/v glycerol, stable for 6 months
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
precipitation, Q-Sepharose Fast Flow column, hydroxylapatite column, Mono-Q GL5/5 column, 4°C
purified to 8.27-fold with a yield of 34.83%, purified by ammonium sulfate fractionation, gel filtration chromatography and anion-exchange chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning and sequencing of gene dcmA; cloning and sequencing of gene METDI2657 located in the 3'-region of the dcmA gene of DCM dehalogenase, RT-PCR expression analysis; cloning and sequencing of gene METDI2658, located in the 3'-region of the dcmA gene of DCM dehalogenase, RT-PCR expression analysis of the gene METDI2658
expression in Escherichia coli
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expression in Escherichia coli BL21 (DE3) from a pET28b(+) expression system
expression in Escherichia coli, Methylobacterium dichloromethanicum and Methylobacterium extorquens
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overexpression in Escherichia coli
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
exposure of cultures growing with methanol to 100 mM NaCl stimulates dichloromethane-dependent induction of dichloromethane dehalogenase activity. However, induction of dichloromethane dehalogenase activity is observed only after addition of dichloromethane, demonstrating that increased salinity is not sufficient to elicit dichloromethane dehalogenase expression
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H8F
-
little effect on the activity, but slower turnover of dibromomethane than wild-type
H8L
-
little effect on the activity, but slower turnover of dibromomethane than wild-type
H8Y
-
little effect on the activity, but slower turnover of dibromomethane than wild-type
S12A
-
drastically reduced activity
S12T
-
drastically reduced activity
V18A
-
lower affinity for GSH and slower turnover of dichlormethane than wild-type
V18I
-
lower affinity for GSH and slower turnover of dichlormethane than wild-type
V18T
-
lower affinity for GSH and slower turnover of dichlormethane than wild-type
W117Y
-
higher KM and lower specific activity for all tested substrates, better heat stability than wild-type
Y6A
-
little effect on the activity
Y6F
-
little effect on the activity
H8F
-
little effect on the activity, but slower turnover of dibromomethane than wild-type
-
H8L
-
little effect on the activity, but slower turnover of dibromomethane than wild-type
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S12A
-
drastically reduced activity
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V18A
-
lower affinity for GSH and slower turnover of dichlormethane than wild-type
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V18I
-
lower affinity for GSH and slower turnover of dichlormethane than wild-type
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V18T
-
lower affinity for GSH and slower turnover of dichlormethane than wild-type
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W117Y
-
higher KM and lower specific activity for all tested substrates, better heat stability than wild-type
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Y6A
-
little effect on the activity
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Y6F
-
little effect on the activity
-
additional information