Functional identification of triterpene methyltransferases from Botryococcus braunii race B
Niehaus, T.D.; Kinison, S.; Okada, S.; Yeo, Y.S.; Bell, S.A.; Cui, P.; Devarenne, T.P.; Chappell, J.; J. Biol. Chem. 287, 8163-8173 (2012) 
Data extracted from this reference:
Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
2.1.1.262
overexpression of TMT-1 and TMT-2 in Saccharomyces cerevisiae microsomes leading to mono- and dimethylated products, co-expression of various TMT and SMT-like genes in Saccharomyces cerevisiae strain TN-7
Botryococcus braunii
2.1.1.263
overexpression of TMT-3 in Saccharomyces cerevisiae microsomes leading to mono- and dimethylated products, co-expression of various TMT and SMT-like genes in Saccharomyces cerevisiae strain TN-7
Botryococcus braunii
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.1.1.262
additional information
-
additional information
Michaelis-Menten enzyme kinetics for TMT-1 and TMT-2
Botryococcus braunii
2.1.1.263
additional information
-
additional information
Michaelis-Menten enzyme kinetics for TMT-3
Botryococcus braunii
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
2.1.1.262
membrane
-
Botryococcus braunii
16020
-
2.1.1.263
membrane
-
Botryococcus braunii
16020
-
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
2.1.1.262
2 S-adenosyl-L-methionine + squalene
Botryococcus braunii
overall reaction
2 S-adenosyl-L-homocysteine + 3,22-dimethyl-1,2,23,24-tetradehydro-2,3,22,23-tetrahydrosqualene
-
-
ir
2.1.1.262
2 S-adenosyl-L-methionine + squalene
Botryococcus braunii race B
overall reaction
2 S-adenosyl-L-homocysteine + 3,22-dimethyl-1,2,23,24-tetradehydro-2,3,22,23-tetrahydrosqualene
-
-
ir
2.1.1.262
S-adenosyl-L-methionine + 3-methyl-1,2-didehydro-2,3-dihydrosqualene
Botryococcus braunii
-
S-adenosyl-L-homocysteine + 3,22-dimethyl-1,2,23,24-tetradehydro-2,3,22,23-tetrahydrosqualene
-
-
?
2.1.1.262
S-adenosyl-L-methionine + squalene
Botryococcus braunii
-
S-adenosyl-L-homocysteine + 3-methyl-1,2-didehydro-2,3-dihydrosqualene
-
-
?
2.1.1.263
2 S-adenosyl-L-methionine + C30 botryococcene
Botryococcus braunii
overall reaction
2 S-adenosyl-L-homocysteine + 3,20-dimethyl-1,2,21,22-tetradehydro-2,3,20,21-tetrahydrobotryococcene
-
-
ir
2.1.1.263
S-adenosyl-L-methionine + 20-methyl-21,22-didehydro-20,21-dihydrobotryococcene
Botryococcus braunii
-
S-adenosyl-L-homocysteine + 3,20-dimethyl-1,2,21,22-tetradehydro-2,3,20,21-tetrahydrobotryococcene
-
-
?
2.1.1.263
S-adenosyl-L-methionine + 3-methyl-1,2-didehydro-2,3-dihydrobotryococcene
Botryococcus braunii
-
S-adenosyl-L-homocysteine + 3,20-dimethyl-1,2,21,22-tetradehydro-2,3,20,21-tetrahydrobotryococcene
-
-
?
2.1.1.263
S-adenosyl-L-methionine + C30 botryococcene
Botryococcus braunii
-
S-adenosyl-L-homocysteine + 20-methyl-21,22-didehydro-20,21-dihydrobotryococcene
-
-
?
2.1.1.263
S-adenosyl-L-methionine + C30 botryococcene
Botryococcus braunii
-
S-adenosyl-L-homocysteine + 3-methyl-1,2-didehydro-2,3-dihydrobotryococcene
-
-
?
Organism
EC Number
Organism
UniProt
Commentary
Textmining
1.3.1.96
Botryococcus braunii
-
race B
-
1.3.1.97
Botryococcus braunii
-
race B
-
2.1.1.262
Botryococcus braunii
-
-
-
2.1.1.262
Botryococcus braunii race B
-
-
-
2.1.1.263
Botryococcus braunii
-
-
-
2.1.1.263
Botryococcus braunii race B
-
-
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.1.1.262
8.56
-
recombinant TMT-1, pH 7.5, 37°C
Botryococcus braunii
2.1.1.262
143.7
-
recombinant TMT-2, pH 7.5, 37°C
Botryococcus braunii
2.1.1.263
7.25
-
recombinant enzyme, pH 7.5, 37°C
Botryococcus braunii
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
1.3.1.96
additional information
SSL-2 cannot efficiently use farnesyl diphosphate as a substrate
720061
Botryococcus braunii
?
-
-
-
?
1.3.1.96
additional information
SSL-2 cannot efficiently use farnesyl diphosphate as a substrate
720061
Botryococcus braunii race B
?
-
-
-
?
1.3.1.96
presqualene diphosphate + NADPH + H+
-
720061
Botryococcus braunii
squalene + diphosphate + NADP+
-
-
-
ir
1.3.1.96
presqualene diphosphate + NADPH + H+
-
720061
Botryococcus braunii race B
squalene + diphosphate + NADP+
-
-
-
ir
1.3.1.97
presqualene diphosphate + NADPH + H+
-
720061
Botryococcus braunii
C30 botryococcene + NADP+ + diphosphate
-
-
-
?
1.3.1.97
presqualene diphosphate + NADPH + H+
-
720061
Botryococcus braunii race B
C30 botryococcene + NADP+ + diphosphate
-
-
-
?
2.1.1.262
2 S-adenosyl-L-methionine + squalene
overall reaction
720061
Botryococcus braunii
2 S-adenosyl-L-homocysteine + 3,22-dimethyl-1,2,23,24-tetradehydro-2,3,22,23-tetrahydrosqualene
-
-
-
ir
2.1.1.262
2 S-adenosyl-L-methionine + squalene
overall reaction
720061
Botryococcus braunii race B
2 S-adenosyl-L-homocysteine + 3,22-dimethyl-1,2,23,24-tetradehydro-2,3,22,23-tetrahydrosqualene
-
-
-
ir
2.1.1.262
additional information
mono- and dimethylated products with terminal carbon C-3 as the atomic acceptor sites for the methyl additions to squalene, NMR analysis, overview TMT-1 and TMT-2 show only low activity with C30 botryococcene
720061
Botryococcus braunii
?
-
-
-
?
2.1.1.262
additional information
mono- and dimethylated products with terminal carbon C-3 as the atomic acceptor sites for the methyl additions to squalene, NMR analysis, overview TMT-1 and TMT-2 show only low activity with C30 botryococcene
720061
Botryococcus braunii race B
?
-
-
-
?
2.1.1.262
S-adenosyl-L-methionine + 3-methyl-1,2-didehydro-2,3-dihydrosqualene
-
720061
Botryococcus braunii
S-adenosyl-L-homocysteine + 3,22-dimethyl-1,2,23,24-tetradehydro-2,3,22,23-tetrahydrosqualene
-
-
-
?
2.1.1.262
S-adenosyl-L-methionine + squalene
-
720061
Botryococcus braunii
S-adenosyl-L-homocysteine + 3-methyl-1,2-didehydro-2,3-dihydrosqualene
-
-
-
?
2.1.1.263
2 S-adenosyl-L-methionine + C30 botryococcene
overall reaction
720061
Botryococcus braunii
2 S-adenosyl-L-homocysteine + 3,20-dimethyl-1,2,21,22-tetradehydro-2,3,20,21-tetrahydrobotryococcene
-
-
-
ir
2.1.1.263
additional information
mono- and dimethylated products with terminal carbons C-22/C-20 as the atomic acceptor sites for the methyl additions to botryococcene, NMR analysis, overview. TMT-3 shows only low activity with squalene
720061
Botryococcus braunii
?
-
-
-
?
2.1.1.263
S-adenosyl-L-methionine + 20-methyl-21,22-didehydro-20,21-dihydrobotryococcene
-
720061
Botryococcus braunii
S-adenosyl-L-homocysteine + 3,20-dimethyl-1,2,21,22-tetradehydro-2,3,20,21-tetrahydrobotryococcene
-
-
-
?
2.1.1.263
S-adenosyl-L-methionine + 3-methyl-1,2-didehydro-2,3-dihydrobotryococcene
-
720061
Botryococcus braunii
S-adenosyl-L-homocysteine + 3,20-dimethyl-1,2,21,22-tetradehydro-2,3,20,21-tetrahydrobotryococcene
-
-
-
?
2.1.1.263
S-adenosyl-L-methionine + C30 botryococcene
-
720061
Botryococcus braunii
S-adenosyl-L-homocysteine + 20-methyl-21,22-didehydro-20,21-dihydrobotryococcene
-
-
-
?
2.1.1.263
S-adenosyl-L-methionine + C30 botryococcene
-
720061
Botryococcus braunii
S-adenosyl-L-homocysteine + 3-methyl-1,2-didehydro-2,3-dihydrobotryococcene
-
-
-
?
Synonyms
EC Number
Synonyms
Commentary
Organism
1.3.1.96
squalene synthase-like 2
-
Botryococcus braunii
1.3.1.96
SSL-2
-
Botryococcus braunii
1.3.1.97
SSL-3
-
Botryococcus braunii
2.1.1.262
triterpene methyltransferase
-
Botryococcus braunii
2.1.1.263
TMT-3
-
Botryococcus braunii
2.1.1.263
triterpene methyltransferase-3
-
Botryococcus braunii
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.1.1.262
37
-
assay at
Botryococcus braunii
2.1.1.263
37
-
assay at
Botryococcus braunii
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.1.1.262
7.5
-
assay at
Botryococcus braunii
2.1.1.263
7.5
-
assay at
Botryococcus braunii
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
2.1.1.262
S-adenosyl-L-methionine
-
Botryococcus braunii
2.1.1.263
S-adenosyl-L-methionine
-
Botryococcus braunii
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.1.262
overexpression of TMT-1 and TMT-2 in Saccharomyces cerevisiae microsomes leading to mono- and dimethylated products, co-expression of various TMT and SMT-like genes in Saccharomyces cerevisiae strain TN-7
Botryococcus braunii
2.1.1.263
overexpression of TMT-3 in Saccharomyces cerevisiae microsomes leading to mono- and dimethylated products, co-expression of various TMT and SMT-like genes in Saccharomyces cerevisiae strain TN-7
Botryococcus braunii
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
2.1.1.262
S-adenosyl-L-methionine
-
Botryococcus braunii
2.1.1.263
S-adenosyl-L-methionine
-
Botryococcus braunii
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.1.1.262
additional information
-
additional information
Michaelis-Menten enzyme kinetics for TMT-1 and TMT-2
Botryococcus braunii
2.1.1.263
additional information
-
additional information
Michaelis-Menten enzyme kinetics for TMT-3
Botryococcus braunii
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
2.1.1.262
membrane
-
Botryococcus braunii
16020
-
2.1.1.263
membrane
-
Botryococcus braunii
16020
-
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
2.1.1.262
2 S-adenosyl-L-methionine + squalene
Botryococcus braunii
overall reaction
2 S-adenosyl-L-homocysteine + 3,22-dimethyl-1,2,23,24-tetradehydro-2,3,22,23-tetrahydrosqualene
-
-
ir
2.1.1.262
2 S-adenosyl-L-methionine + squalene
Botryococcus braunii race B
overall reaction
2 S-adenosyl-L-homocysteine + 3,22-dimethyl-1,2,23,24-tetradehydro-2,3,22,23-tetrahydrosqualene
-
-
ir
2.1.1.262
S-adenosyl-L-methionine + 3-methyl-1,2-didehydro-2,3-dihydrosqualene
Botryococcus braunii
-
S-adenosyl-L-homocysteine + 3,22-dimethyl-1,2,23,24-tetradehydro-2,3,22,23-tetrahydrosqualene
-
-
?
2.1.1.262
S-adenosyl-L-methionine + squalene
Botryococcus braunii
-
S-adenosyl-L-homocysteine + 3-methyl-1,2-didehydro-2,3-dihydrosqualene
-
-
?
2.1.1.263
2 S-adenosyl-L-methionine + C30 botryococcene
Botryococcus braunii
overall reaction
2 S-adenosyl-L-homocysteine + 3,20-dimethyl-1,2,21,22-tetradehydro-2,3,20,21-tetrahydrobotryococcene
-
-
ir
2.1.1.263
S-adenosyl-L-methionine + 20-methyl-21,22-didehydro-20,21-dihydrobotryococcene
Botryococcus braunii
-
S-adenosyl-L-homocysteine + 3,20-dimethyl-1,2,21,22-tetradehydro-2,3,20,21-tetrahydrobotryococcene
-
-
?
2.1.1.263
S-adenosyl-L-methionine + 3-methyl-1,2-didehydro-2,3-dihydrobotryococcene
Botryococcus braunii
-
S-adenosyl-L-homocysteine + 3,20-dimethyl-1,2,21,22-tetradehydro-2,3,20,21-tetrahydrobotryococcene
-
-
?
2.1.1.263
S-adenosyl-L-methionine + C30 botryococcene
Botryococcus braunii
-
S-adenosyl-L-homocysteine + 20-methyl-21,22-didehydro-20,21-dihydrobotryococcene
-
-
?
2.1.1.263
S-adenosyl-L-methionine + C30 botryococcene
Botryococcus braunii
-
S-adenosyl-L-homocysteine + 3-methyl-1,2-didehydro-2,3-dihydrobotryococcene
-
-
?
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.1.1.262
8.56
-
recombinant TMT-1, pH 7.5, 37°C
Botryococcus braunii
2.1.1.262
143.7
-
recombinant TMT-2, pH 7.5, 37°C
Botryococcus braunii
2.1.1.263
7.25
-
recombinant enzyme, pH 7.5, 37°C
Botryococcus braunii
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
1.3.1.96
additional information
SSL-2 cannot efficiently use farnesyl diphosphate as a substrate
720061
Botryococcus braunii
?
-
-
-
?
1.3.1.96
additional information
SSL-2 cannot efficiently use farnesyl diphosphate as a substrate
720061
Botryococcus braunii race B
?
-
-
-
?
1.3.1.96
presqualene diphosphate + NADPH + H+
-
720061
Botryococcus braunii
squalene + diphosphate + NADP+
-
-
-
ir
1.3.1.96
presqualene diphosphate + NADPH + H+
-
720061
Botryococcus braunii race B
squalene + diphosphate + NADP+
-
-
-
ir
1.3.1.97
presqualene diphosphate + NADPH + H+
-
720061
Botryococcus braunii
C30 botryococcene + NADP+ + diphosphate
-
-
-
?
1.3.1.97
presqualene diphosphate + NADPH + H+
-
720061
Botryococcus braunii race B
C30 botryococcene + NADP+ + diphosphate
-
-
-
?
2.1.1.262
2 S-adenosyl-L-methionine + squalene
overall reaction
720061
Botryococcus braunii
2 S-adenosyl-L-homocysteine + 3,22-dimethyl-1,2,23,24-tetradehydro-2,3,22,23-tetrahydrosqualene
-
-
-
ir
2.1.1.262
2 S-adenosyl-L-methionine + squalene
overall reaction
720061
Botryococcus braunii race B
2 S-adenosyl-L-homocysteine + 3,22-dimethyl-1,2,23,24-tetradehydro-2,3,22,23-tetrahydrosqualene
-
-
-
ir
2.1.1.262
additional information
mono- and dimethylated products with terminal carbon C-3 as the atomic acceptor sites for the methyl additions to squalene, NMR analysis, overview TMT-1 and TMT-2 show only low activity with C30 botryococcene
720061
Botryococcus braunii
?
-
-
-
?
2.1.1.262
additional information
mono- and dimethylated products with terminal carbon C-3 as the atomic acceptor sites for the methyl additions to squalene, NMR analysis, overview TMT-1 and TMT-2 show only low activity with C30 botryococcene
720061
Botryococcus braunii race B
?
-
-
-
?
2.1.1.262
S-adenosyl-L-methionine + 3-methyl-1,2-didehydro-2,3-dihydrosqualene
-
720061
Botryococcus braunii
S-adenosyl-L-homocysteine + 3,22-dimethyl-1,2,23,24-tetradehydro-2,3,22,23-tetrahydrosqualene
-
-
-
?
2.1.1.262
S-adenosyl-L-methionine + squalene
-
720061
Botryococcus braunii
S-adenosyl-L-homocysteine + 3-methyl-1,2-didehydro-2,3-dihydrosqualene
-
-
-
?
2.1.1.263
2 S-adenosyl-L-methionine + C30 botryococcene
overall reaction
720061
Botryococcus braunii
2 S-adenosyl-L-homocysteine + 3,20-dimethyl-1,2,21,22-tetradehydro-2,3,20,21-tetrahydrobotryococcene
-
-
-
ir
2.1.1.263
additional information
mono- and dimethylated products with terminal carbons C-22/C-20 as the atomic acceptor sites for the methyl additions to botryococcene, NMR analysis, overview. TMT-3 shows only low activity with squalene
720061
Botryococcus braunii
?
-
-
-
?
2.1.1.263
S-adenosyl-L-methionine + 20-methyl-21,22-didehydro-20,21-dihydrobotryococcene
-
720061
Botryococcus braunii
S-adenosyl-L-homocysteine + 3,20-dimethyl-1,2,21,22-tetradehydro-2,3,20,21-tetrahydrobotryococcene
-
-
-
?
2.1.1.263
S-adenosyl-L-methionine + 3-methyl-1,2-didehydro-2,3-dihydrobotryococcene
-
720061
Botryococcus braunii
S-adenosyl-L-homocysteine + 3,20-dimethyl-1,2,21,22-tetradehydro-2,3,20,21-tetrahydrobotryococcene
-
-
-
?
2.1.1.263
S-adenosyl-L-methionine + C30 botryococcene
-
720061
Botryococcus braunii
S-adenosyl-L-homocysteine + 20-methyl-21,22-didehydro-20,21-dihydrobotryococcene
-
-
-
?
2.1.1.263
S-adenosyl-L-methionine + C30 botryococcene
-
720061
Botryococcus braunii
S-adenosyl-L-homocysteine + 3-methyl-1,2-didehydro-2,3-dihydrobotryococcene
-
-
-
?
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.1.1.262
37
-
assay at
Botryococcus braunii
2.1.1.263
37
-
assay at
Botryococcus braunii
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.1.1.262
7.5
-
assay at
Botryococcus braunii
2.1.1.263
7.5
-
assay at
Botryococcus braunii
General Information
EC Number
General Information
Commentary
Organism
2.1.1.262
metabolism
TMT-1 and TMT-2 exhibit preference for squalene as the methyl acceptor substrate
Botryococcus braunii
2.1.1.263
metabolism
TMT-3 shows a striking preference for botryococcene as its methyl acceptor substrate
Botryococcus braunii
General Information (protein specific)
EC Number
General Information
Commentary
Organism
2.1.1.262
metabolism
TMT-1 and TMT-2 exhibit preference for squalene as the methyl acceptor substrate
Botryococcus braunii
2.1.1.263
metabolism
TMT-3 shows a striking preference for botryococcene as its methyl acceptor substrate
Botryococcus braunii