Information on EC 2.1.1.165 - methyl halide transferase

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

EC NUMBER
COMMENTARY hide
2.1.1.165
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RECOMMENDED NAME
GeneOntology No.
methyl halide transferase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
S-adenosyl-L-methionine + iodide = S-adenosyl-L-homocysteine + methyl iodide
show the reaction diagram
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
methylhalides biosynthesis (plants)
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SYSTEMATIC NAME
IUBMB Comments
S-adenosylmethionine:I- methyltransferase
This enzyme contributes to the methyl halide emissions from Arabidopsis [6].
CAS REGISTRY NUMBER
COMMENTARY hide
129877-08-3
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
chinensis
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
strain MRCD 19
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Manually annotated by BRENDA team
strain MRCD 19
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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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
S-adenosyl-L-methionine + bromide
S-adenosyl-L-homocysteine + methyl bromide
show the reaction diagram
S-adenosyl-L-methionine + chloride
S-adenosyl-L-homocysteine + methyl chloride
show the reaction diagram
S-adenosyl-L-methionine + iodide
S-adenosyl-L-homocysteine + methyl iodide
show the reaction diagram
additional information
?
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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
S-adenosyl-L-methionine + chloride
S-adenosyl-L-homocysteine + methyl chloride
show the reaction diagram
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
S-adenosyl-L-methionine
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ammonium sulfate
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the activity of the recombinant methylase in 0.5 M and 1.0 M ammonium sulfate is about 1.25fold higher than in the absence of ammonium sulfate
additional information
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various metal ions have no significant effect on methyl iodide production, suggesting that the enzyme does not require metal ions
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
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1 mM, 16% inhibition, production of methyl iodide
bisulfide
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chloride
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competitive inhibition of methyl iodide formation
CN-
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100 mM, complete inhibition, production of methyl iodide
CoCl2
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1 mM, 18% inhibition, production of methyl iodide
dithiothreitol
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5 mM, 28% inhibition, production of methyl iodide
EDTA
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5 mM, 32% inhibition, production of methyl iodide
HS-
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10 mM, 84% inhibition, production of methyl iodide
Iodide
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above 25 mM
MgSO4
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1 mM, 25% inhibition, production of methyl iodide
monoiodoacetate
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5 mM, 28% inhibition, production of methyl iodide
NaN3
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5 mM, 32% inhibition, production of methyl iodide
NiCl2
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1 mM, 38% inhibition, production of methyl iodide
PCMB
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0.5 mM, 30% inhibition, production of methyl iodide
S-adenosyl-L-homocysteine
SCN-
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10 mM, 60% inhibition, production of methyl iodide
ZnCl2
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1 mM, 14% inhibition, production of methyl iodide
additional information
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activity is not inhibited by high iodide concentrations
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Urea
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activity in 1, 2, and 3 M urea is about 1.75-, 2-, and 1.5-fold higher than in the absence of urea, respectively
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5 - 177.3
bromide
85 - 1657
chloride
0.25 - 63
Iodide
0.0045 - 0.23
S-adenosyl-L-methionine
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.35
bisulfide
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pH 7.5, 22C
0.032
S-adenosyl-L-homocysteine
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pH 7.5, 22C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.000000547
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0.0000058
production of methyl iodide
0.00009
production of methyl iodide
0.000555
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partially purified enzyme, synthesis of methyl bromide
0.0018
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0.3
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purified enzyme, substrate iodide
205
production of methyl iodide
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 7
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methylation of iodide
6.2 - 6.8
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methyl chloride production
6.2
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methyl bromide production
6.8 - 7.5
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methyl iodide production
6.8
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assay at
7 - 7.5
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7
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assay at
7.5 - 7.6
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synthesis of methyl bromide
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.4 - 10
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pH 5.4: about 50% of maximal activity, pH 10.0: about 80% of maximal activity
6 - 9
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pH 6.0: about 40% of maximal activity, pH 9.0: about 35% of maximal activity, at pH greater than 9.2 no methyl bromide synthesis detected
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.8
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chromatofocusing
5.1
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calculated from sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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the fungus is cultured in undisturbed glucose mycological peptone liquid medium
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20000 - 25000
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gel filtration
29000
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gel filtration
29500
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
- 80C, enzyme after the first gel filtration purification step, in 25 mM Tris acetate, pH 7.4, 10% glycerol, and 14 mM 2-mercaptoethanol, stable for over 2 months
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-20C to 4C, partially purified enzyme, complete loss of activity overnight, also in the presence of protease inhibitors
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-20C, enzyme forms an aggregate with molecular mass of approximately 500000 Da
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-20C, enzyme in cell extract is unstable and loses activities almost completely upon storage even if dithioerythritol, EDTA, protease inhibitor or glycerol are added to the extracts
-20C, purified protein stored in a buffer (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 1 mM DTT, 30% glycerol), after 15 days, the recombinant protein AtHOL1 retains 40% of the iodide methyltransferase activity
-20C, purified protein stored in a buffer (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 1 mM DTT, 30% glycerol), after 15 days, the recombinant protein AtHOL1 retains 60% of the iodide methyltransferase activity
-20C, purified protein stored in a buffer (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 1 mM DTT, 30% glycerol), after 15 days, the recombinant protein AtHOL1 retains 90% of the iodide methyltransferase activity
-80C, after affinity chromatography, the halide/bisulfide methyltransferase becomes extremely labile losing all activity after overnight storage
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20C, enzyme after the affinity chromatography purification step, 25 mM Tris acetate, pH 7.4, 14 mM 2-mercaptoethanol, and 30% glycerol, 12% remaining activity after 48 h
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4C, enzyme after anion exchange purification step, in 25 mM Tris acetate, pH 7.4, 14 mM 2-mercaptoethanol, and 175 mM NaCl, more than 70% remaining activity after 24 h and 55% after 48 h
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4C, enzyme in cell extract is unstable and loses activities almost completely upon storage even if dithioerythritol, EDTA, protease inhibitor or glycerol are added to the extracts
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
gel filtration, anion exchange chromatography, and affinity chromatography on adenosine-agarose
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native enzyme 2700fold to homogeneity by ammonium sulfate fractionation, gel filtration, adenosine affinity chromatography, and a second step of gel filtration
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the difficulty of solubilization of this membrane-bound labile enzyme is the greatest obstacle to its purification
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the purification is achieved by an 80 to 100% ammonium sulfate precipitation step followed by a high-performance liquid chromatography gel filtration step on a 60 cm by 2.15 cm preparative Bio-Sil SEC-250 column. The column is eluted with 10 mM phosphate buffer, pH 7.0
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
expression in Escherichia coli
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synthesis of all putative methyl halide transferase from the NCBI sequence database and assay of methyl halide production in Escherichia coli
synthesis of all putative methyl halide transferases from the NCBI sequence database and assay of methyl halide production in Escherichia coli
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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the disparity between the observed (22500 Da) and calculated molecular mass (25761 Da) suggests that the methylase undergoes posttranslational cleavage, possibly during purification
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis