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Literature summary extracted from

  • Okuyama, M.; Kitamura, M.; Hondoh, H.; Kang, M.S.; Mori, H.; Kimura, A.; Tanaka, I.; Yao, M.
    Catalytic mechanism of retaining alpha-galactosidase belonging to glycoside hydrolase family 97 (2009), J. Mol. Biol., 392, 1232-1241.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.22 expressed in Escherichia coli MV1184 cells Bacteroides thetaiotaomicron

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.22 hanging drop vapor diffusion method, using 100 mM MES-NaOH buffer, pH 6.2, containing 12% (w/v) PEG (polyethylene glycol) 6000 or 100 mM MES-NaOH buffer containing 10% (w/v) PEG 10000, at 20°C Bacteroides thetaiotaomicron

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.22 D415G inactive Bacteroides thetaiotaomicron
3.2.1.22 D415N the catalytic activity of the mutant enzyme E470Q is significantly decreased (kcat is 100000 lower than that of the wild type enzyme whereas the Km values of both D415N and wild type enzymes are roughly similar.) Bacteroides thetaiotaomicron
3.2.1.22 E470Q the catalytic activity of the mutant enzyme E470Q is significantly decreased (kcat is 28000 lower than that of the wild type enzyme whereas the Km values of both D415N and wild type enzymes are the same) Bacteroides thetaiotaomicron

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.22 0.19
-
 4-nitrophenyl alpha-D-galactoside mutant enzyme D415N, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C Bacteroides thetaiotaomicron
3.2.1.22 0.31
-
 4-nitrophenyl alpha-D-galactoside wild type enzyme, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C Bacteroides thetaiotaomicron
3.2.1.22 0.32
-
 4-nitrophenyl alpha-D-galactoside mutant enzyme E470Q, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C Bacteroides thetaiotaomicron

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.22 Bacteroides thetaiotaomicron Q8A6L0 strain ATCC 29148
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.22 Ni-chelating Sepharose column chromatography and DEAE-Sepharose column chromatography Bacteroides thetaiotaomicron

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.22 1-azido-1-deoxy-beta-D-galactopyranoside + H2O
-
 Bacteroides thetaiotaomicron ?
-
 ?
3.2.1.22 4-nitrophenyl alpha-D-galactoside + H2O
-
 Bacteroides thetaiotaomicron 4-nitrophenol + alpha-D-galactose
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.22 GH97b
-
 Bacteroides thetaiotaomicron
3.2.1.22 retaining alpha-galactosidase
-
 Bacteroides thetaiotaomicron

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.22 0.00218
-
 4-nitrophenyl alpha-D-galactoside mutant enzyme D415N, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C Bacteroides thetaiotaomicron
3.2.1.22 0.0684
-
 4-nitrophenyl alpha-D-galactoside mutant enzyme E470Q, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C Bacteroides thetaiotaomicron
3.2.1.22 244
-
 4-nitrophenyl alpha-D-galactoside wild type enzyme, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C Bacteroides thetaiotaomicron

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.22 0.0115
-
 4-nitrophenyl alpha-D-galactoside mutant enzyme D415N, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C Bacteroides thetaiotaomicron
3.2.1.22 0.213
-
 4-nitrophenyl alpha-D-galactoside mutant enzyme E470Q, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C Bacteroides thetaiotaomicron
3.2.1.22 778
-
 4-nitrophenyl alpha-D-galactoside wild type enzyme, in 40 mM sodium phosphate buffer, pH 6.0, at 35°C Bacteroides thetaiotaomicron