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Literature summary extracted from

  • Comfort, D.A.; Bobrov, K.S.; Ivanen, D.R.; Shabalin, K.A.; Harris, J.M.; Kulminskaya, A.A.; Brumer, H.; Kelly, R.M.
    Biochemical analysis of Thermotoga maritima GH36 alpha-galactosidase (TmGalA) confirms the mechanistic commonality of clan GH-D glycoside hydrolases (2007), Biochemistry, 46, 3319-3330.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.22 expressed in Escherichia coli BL-21(DE3) cells Thermotoga maritima

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.22 D220A exhibits less than 1% of the wild-type enzyme activity Thermotoga maritima
3.2.1.22 D327A exhibits less than 1% of the wild-type enzyme activity Thermotoga maritima
3.2.1.22 D327G mutant has a 200-800fold lower catalytic rate on aryl galactosides relative to the wild type enzyme Thermotoga maritima
3.2.1.22 D387A exhibits less than 1% of the wild-type enzyme activity Thermotoga maritima
3.2.1.22 D387G mutant is 1500fold catalytically slower than the wild type enzyme on p-nitrophenyl alpha-D-galactopyranoside, and exhibits higher activity with increasing pH Thermotoga maritima

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.22 0.037
-
 2,4-dinitrophenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.039
-
 beta-naphthyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.048
-
 4-O-methylphenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.08
-
 4-nitrophenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.083
-
 4-nitrophenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.086
-
 3,4-dimethylphenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.088
-
 4-O-methylphenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.089
-
 3,4-dimethylphenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.096
-
 3-nitrophenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.104
-
 4-bromophenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.111
-
 4-bromophenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.133
-
 2-nitrophenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.135
-
 2,4-dinitrophenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.14
-
 3-nitrophenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.143
-
 2-nitrophenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.143
-
 2,5-dinitrophenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.176
-
 phenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.225
-
 beta-naphthyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.243
-
 2,5-dinitrophenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.324
-
 phenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.22 Thermotoga maritima O33835
-
-
3.2.1.22 Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 O33835
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.22
-
 Thermotoga maritima

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.22 2,4-dinitrophenyl alpha-D-galactoside + H2O
-
 Thermotoga maritima 2,4-dinitrophenol + alpha-D-galactose
-
 ?
3.2.1.22 2,4-dinitrophenyl alpha-D-galactoside + H2O
-
 Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 2,4-dinitrophenol + alpha-D-galactose
-
 ?
3.2.1.22 2,5-dinitrophenyl alpha-D-galactoside + H2O
-
 Thermotoga maritima 2,5-dinitrophenol + alpha-D-galactose
-
 ?
3.2.1.22 2,5-dinitrophenyl alpha-D-galactoside + H2O
-
 Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 2,5-dinitrophenol + alpha-D-galactose
-
 ?
3.2.1.22 2-nitrophenyl alpha-D-galactoside + H2O
-
 Thermotoga maritima 2-nitrophenol + alpha-D-galactose
-
 ?
3.2.1.22 2-nitrophenyl alpha-D-galactoside + H2O
-
 Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 2-nitrophenol + alpha-D-galactose
-
 ?
3.2.1.22 3,4-dimethylphenyl alpha-D-galactoside + H2O
-
 Thermotoga maritima 3,4-dimethylphenol + alpha-D-galactose
-
 ?
3.2.1.22 3,4-dimethylphenyl alpha-D-galactoside + H2O
-
 Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 3,4-dimethylphenol + alpha-D-galactose
-
 ?
3.2.1.22 3-nitrophenyl alpha-D-galactoside + H2O
-
 Thermotoga maritima 3-nitrophenol + alpha-D-galactose
-
 ?
3.2.1.22 4-bromophenyl alpha-D-galactoside + H2O
-
 Thermotoga maritima 4-bromophenol + alpha-D-galactose
-
 ?
3.2.1.22 4-methoxyphenyl alpha-D-galactoside + H2O
-
 Thermotoga maritima 4-methoxyphenol + alpha-D-galactose
-
 ?
3.2.1.22 4-nitrophenyl alpha-D-galactoside + H2O
-
 Thermotoga maritima 4-nitrophenol + alpha-D-galactose
-
 ?
3.2.1.22 beta-naphthyl alpha-D-galactoside + H2O
-
 Thermotoga maritima beta-naphthol + alpha-D-galactose
-
 ?
3.2.1.22 additional information no detectable enzymatic activity on xylo-, fuco-, manno-, or glucopyranosides Thermotoga maritima ?
-
 ?
3.2.1.22 additional information no detectable enzymatic activity on xylo-, fuco-, manno-, or glucopyranosides Thermotoga maritima MSB8 / DSM 3109 / ATCC 43589 ?
-
 ?
3.2.1.22 phenyl alpha-D-galactoside + H2O
-
 Thermotoga maritima phenol + alpha-D-galactose
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.22 GALA
-
 Thermotoga maritima
3.2.1.22 GH36 alpha-galactosidase
-
 Thermotoga maritima

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.22 89.8
-
 melting temperature of the wild type enzyme Thermotoga maritima

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.22 additional information
-
 phenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.01
-
 4-O-methylphenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.01
-
 3,4-dimethylphenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.016
-
 4-bromophenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.021
-
 beta-naphthyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.027
-
 phenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.03 0.55 4-O-methylphenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.03
-
 4-nitrophenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.035
-
 2-nitrophenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.042
-
 2,5-dinitrophenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.042
-
 3-nitrophenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.052 2.1 3,4-dimethylphenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.085
-
 2,4-dinitrophenyl alpha-D-galactoside mutant enzyme D327G, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.41
-
 2,4-dinitrophenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 0.52
-
 2-nitrophenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 5.71
-
 phenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 6.26
-
 beta-naphthyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 6.69
-
 4-O-methylphenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 8.58
-
 4-nitrophenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 10.39
-
 3,4-dimethylphenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 10.5
-
 3-nitrophenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 11.2
-
 2,5-dinitrophenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 12.23
-
 2,4-dinitrophenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 12.58
-
 4-bromophenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima
3.2.1.22 13.67
-
 2-nitrophenyl alpha-D-galactoside wild type enzyme, in 200 mM sodium citrate buffer (pH 5.0), at 37°C Thermotoga maritima