BRENDA - Enzyme Database show
show all sequences of 2.1.1.165

Purification and characterization of a novel methyltransferase responsible for biosynthesis of halomethanes and methanethiol in Brassica oleracea

Attieh, J.M.; Hanson, A.D.; Saini, H.S.; J. Biol. Chem. 270, 9250-9257 (1995)

Data extracted from this reference:

Inhibitors
Inhibitors
Commentary
Organism
Structure
bisulfide
-
Brassica oleracea
S-adenosyl-L-homocysteine
competitive with respect to S-adenosyl-L-methionine, noncompetitive to iodide
Brassica oleracea
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.03
-
S-adenosyl-L-methionine
pH 7.5, 22°C, cosubstrate: iodide
Brassica oleracea
1.3
-
Iodide
pH 7.5, 22°C
Brassica oleracea
29
-
bromide
pH 7.5, 22°C
Brassica oleracea
85
-
chloride
pH 7.5, 22°C
Brassica oleracea
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
28000
-
1 * 28000, SDS-PAFE
Brassica oleracea
29500
-
gel filtration
Brassica oleracea
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Brassica oleracea
-
-
-
Purification (Commentary)
Commentary
Organism
gel filtration, anion exchange chromatography, and affinity chromatography on adenosine-agarose
Brassica oleracea
Source Tissue
Source Tissue
Commentary
Organism
Textmining
leaf
-
Brassica oleracea
-
Storage Stability
Storage Stability
Organism
- 80°C, 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
Brassica oleracea
-80°C, after affinity chromatography, the halide/bisulfide methyltransferase becomes extremely labile losing all activity after overnight storage
Brassica oleracea
20°C, 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
Brassica oleracea
4°C, 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
Brassica oleracea
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
additional information
the bifunctional enzyme also shows activity of EC 2.1.1.9
680600
Brassica oleracea
?
-
-
-
-
S-adenosyl-L-methionine + bromide
kcat/KM for bromide is 12529fold lower than kcat/Km for iodide
680600
Brassica oleracea
S-adenosyl-L-homocysteine + methyl bromide
-
-
-
?
S-adenosyl-L-methionine + chloride
kcat/KM for chloride is 19065fold lower than kcat/Km for iodide
680600
Brassica oleracea
S-adenosyl-L-homocysteine + methyl chloride
-
-
-
?
S-adenosyl-L-methionine + iodide
iodide is the preferred substrate
680600
Brassica oleracea
S-adenosyl-L-homocysteine + methyl iodide
-
-
-
?
Subunits
Subunits
Commentary
Organism
monomer
1 * 28000, SDS-PAFE
Brassica oleracea
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
22
-
assay at
Brassica oleracea
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
5.5
7
methylation of iodide
Brassica oleracea
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.032
-
S-adenosyl-L-homocysteine
pH 7.5, 22°C
Brassica oleracea
1.35
-
bisulfide
pH 7.5, 22°C
Brassica oleracea
pI Value
Organism
Commentary
pI Value Maximum
pI Value
Brassica oleracea
chromatofocusing
-
4.8
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
bisulfide
-
Brassica oleracea
S-adenosyl-L-homocysteine
competitive with respect to S-adenosyl-L-methionine, noncompetitive to iodide
Brassica oleracea
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.032
-
S-adenosyl-L-homocysteine
pH 7.5, 22°C
Brassica oleracea
1.35
-
bisulfide
pH 7.5, 22°C
Brassica oleracea
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.03
-
S-adenosyl-L-methionine
pH 7.5, 22°C, cosubstrate: iodide
Brassica oleracea
1.3
-
Iodide
pH 7.5, 22°C
Brassica oleracea
29
-
bromide
pH 7.5, 22°C
Brassica oleracea
85
-
chloride
pH 7.5, 22°C
Brassica oleracea
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
28000
-
1 * 28000, SDS-PAFE
Brassica oleracea
29500
-
gel filtration
Brassica oleracea
Purification (Commentary) (protein specific)
Commentary
Organism
gel filtration, anion exchange chromatography, and affinity chromatography on adenosine-agarose
Brassica oleracea
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
leaf
-
Brassica oleracea
-
Storage Stability (protein specific)
Storage Stability
Organism
- 80°C, 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
Brassica oleracea
-80°C, after affinity chromatography, the halide/bisulfide methyltransferase becomes extremely labile losing all activity after overnight storage
Brassica oleracea
20°C, 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
Brassica oleracea
4°C, 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
Brassica oleracea
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
additional information
the bifunctional enzyme also shows activity of EC 2.1.1.9
680600
Brassica oleracea
?
-
-
-
-
S-adenosyl-L-methionine + bromide
kcat/KM for bromide is 12529fold lower than kcat/Km for iodide
680600
Brassica oleracea
S-adenosyl-L-homocysteine + methyl bromide
-
-
-
?
S-adenosyl-L-methionine + chloride
kcat/KM for chloride is 19065fold lower than kcat/Km for iodide
680600
Brassica oleracea
S-adenosyl-L-homocysteine + methyl chloride
-
-
-
?
S-adenosyl-L-methionine + iodide
iodide is the preferred substrate
680600
Brassica oleracea
S-adenosyl-L-homocysteine + methyl iodide
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
monomer
1 * 28000, SDS-PAFE
Brassica oleracea
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
22
-
assay at
Brassica oleracea
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
5.5
7
methylation of iodide
Brassica oleracea
pI Value (protein specific)
Organism
Commentary
pI Value Maximum
pI Value
Brassica oleracea
chromatofocusing
-
4.8
Other publictions for EC 2.1.1.165
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
696997
Itoh
Involvement of S-adenosylmethi ...
Raphanus sativus
BMC Plant Biol.
9
116
2009
-
-
1
-
-
-
-
4
-
-
2
1
-
1
-
-
1
-
-
1
-
-
5
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
4
-
-
2
1
-
-
-
1
-
1
-
-
5
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
698496
Bayer
Synthesis of methyl halides fr ...
Batis maritima, Brassica rapa, Burkholderia pseudomallei, Burkholderia thailandensis, Oryza sativa, Paraburkholderia phytofirmans, Paraburkholderia xenovorans, Vitis vinifera
J. Am. Chem. Soc.
131
6508-6515
2009
-
8
8
-
-
-
-
-
-
-
-
-
-
16
-
-
-
-
-
-
-
-
21
-
-
-
-
-
-
-
-
-
-
-
-
-
8
8
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
21
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
698916
Nagatoshi Y
Arabidopsis HARMLESS TO OZONE ...
Arabidopsis thaliana
J. Biol. Chem.
284
19301-19309
2009
-
-
-
-
-
-
-
-
-
-
-
1
-
3
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
700702
Nagatoshi. Y.; Nakamura
-
Characterization of three hali ...
Arabidopsis thaliana
Plant Biotechnol.
24
503-506
2007
-
-
1
-
-
-
-
-
-
-
2
3
-
3
-
-
-
-
-
-
3
3
9
1
-
-
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
3
3
-
-
-
-
-
-
3
3
9
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
679357
Rhew
Genetic control of methyl hali ...
Arabidopsis thaliana
Curr. Biol.
13
1809-1813
2003
-
-
1
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
1
-
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-
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1
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-
-
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1
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-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
695730
Amachi
Bacteria mediate methylation o ...
Rhizobium sp., Rhizobium sp. MRCD 19
Appl. Environ. Microbiol.
67
2718-2722
2001
-
-
-
-
-
-
1
2
-
-
-
2
-
2
-
-
-
-
-
-
1
-
4
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
2
-
-
-
2
-
-
-
-
-
-
1
-
4
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
696802
Ohsawa
Purification and characterizat ...
Pavlova pinguis
Biosci. Biotechnol. Biochem.
65
2397-2404
2001
-
-
-
-
-
-
14
2
-
1
1
-
-
2
-
-
1
-
-
-
1
-
4
1
-
-
-
-
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
14
-
2
-
1
1
-
-
-
-
1
-
-
1
-
4
1
-
-
-
-
1
1
-
1
-
-
-
-
-
-
682595
Ni
Expression of Batis maritima m ...
Batis maritima
Proc. Natl. Acad. Sci. USA
96
3611-3615
1999
1
-
1
-
-
-
1
8
-
1
-
1
-
5
-
-
-
-
-
-
-
-
4
-
-
-
-
-
3
-
-
-
-
-
-
1
-
1
-
-
-
-
-
1
-
8
-
1
-
1
-
-
-
-
-
-
-
-
4
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
677650
Saxena
Biochemical characterization o ...
Phellinus pomaceus
Appl. Environ. Microbiol.
64
2831-2835
1998
-
-
-
-
-
-
1
4
1
-
-
1
-
3
-
-
1
-
-
1
-
1
4
-
-
-
-
-
1
1
-
1
-
-
-
-
-
-
1
-
-
-
-
1
-
4
1
-
-
1
-
-
-
1
-
1
-
1
4
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
682593
Ni
cDNA cloning of Batis maritima ...
Batis maritima
Proc. Natl. Acad. Sci. USA
95
12866-12871
1998
-
-
1
-
1
-
-
-
-
-
3
1
-
3
-
-
1
-
-
1
1
1
4
1
1
-
-
-
1
-
-
1
-
1
-
-
-
1
1
-
1
-
-
-
-
-
-
-
3
1
-
-
-
1
-
1
1
1
4
1
1
-
-
-
1
-
-
1
-
-
-
-
-
-
700660
Itoh
-
Formation and emission of mono ...
Papenfussiella kuromo, Pavlova gyrans, Sargassum horneri
Phytochemistry
45
67-73
1997
-
-
-
-
-
-
1
2
-
-
-
3
-
3
-
-
-
-
-
-
-
4
8
-
1
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
2
-
-
-
3
-
-
-
-
-
-
-
4
8
-
1
-
-
-
1
1
-
-
-
-
-
-
-
-
680600
Attieh
Purification and characterizat ...
Brassica oleracea
J. Biol. Chem.
270
9250-9257
1995
-
-
-
-
-
-
2
4
-
-
2
-
-
2
-
-
1
-
-
1
-
4
4
1
1
-
-
-
1
-
-
-
2
1
-
-
-
-
-
-
-
-
-
2
2
4
-
-
2
-
-
-
-
1
-
1
-
4
4
1
1
-
-
-
1
-
-
1
-
-
-
-
-
-
700704
Saini
-
Biosynthesis of halomethanes a ...
Brassica oleracea
Plant Cell Environ.
18
1027-1033
1995
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
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-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
701215
Wuosmaa
Methyl chloride transferase: a ...
Endocladia muricata
Science
249
160-162
1990
-
-
-
-
-
-
-
2
-
-
1
-
-
4
-
-
1
-
-
-
1
-
4
-
-
-
-
-
1
1
-
-
-
-
-
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-
-
-
-
-
-
-
-
-
2
-
-
1
-
-
-
-
1
-
-
1
-
4
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-