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Literature summary for 3.2.1.23 extracted from

  • Shaikh, F.A.; Randriantsoa, M.; Withers, S.G.
    Mechanistic analysis of the blood group antigen-cleaving endo-beta-galactosidase from Clostridium perfringens (2009), Biochemistry, 48, 8396-8404.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Clostridium perfringens

Protein Variants

Protein Variants Comment Organism
D429A the mutant has wild type activity Clostridium perfringens
D453A inactive Clostridium perfringens
E354A the activity of the E354A mutant with nonactivated natural substrates is 1100fold lower than that of the wild type enzyme, while its activity is only 10fold lower when assayed with 2,4-dinitrophenyl 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-3)-[alpha-L-fucopyranosyl-(1-2)]-beta-D-galactopyranoside Clostridium perfringens
E467A the mutant has wild type activity Clostridium perfringens
E506A inactive Clostridium perfringens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.031
-
2,4-dinitrophenyl 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-3)-[alpha-L-fucopyranosyl-(1-2)]-beta-D-galactopyranoside mutant enzyme E354A, in 20 mM sodium acetate buffer at pH 6.0, 37°C Clostridium perfringens
0.059
-
2,4-dinitrophenyl 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-3)-[alpha-L-fucopyranosyl-(1-2)]-beta-D-galactopyranoside mutant enzyme E467A, in 20 mM sodium acetate buffer at pH 6.0, 37°C Clostridium perfringens
0.061
-
2,4-dinitrophenyl 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-3)-[alpha-L-fucopyranosyl-(1-2)]-beta-D-galactopyranoside mutant enzyme D429A, in 20 mM sodium acetate buffer at pH 6.0, 37°C Clostridium perfringens
0.064
-
2,4-dinitrophenyl 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-3)-[alpha-L-fucopyranosyl-(1-2)]-beta-D-galactopyranoside wild type enzyme, in 20 mM sodium acetate buffer at pH 6.0, 37°C Clostridium perfringens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
88000
-
x * 88000, SDS-PAGE Clostridium perfringens
88150
-
subunit, calculated from amino acid sequence Clostridium perfringens

Organism

Organism UniProt Comment Textmining
Clostridium perfringens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
HisTrap column chromatography and MonoQ column chromatography Clostridium perfringens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,4-dinitrophenyl 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-3)-[alpha-L-fucopyranosyl-(1-2)]-beta-D-galactopyranoside + H2O
-
Clostridium perfringens 2,4-dinitrophenol + 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-3)-[alpha-L-fucopyranosyl-(1-2)]-beta-D-galactopyranoside
-
?
Gal-alpha(1,3)-Gal-beta-2,4-dinitrophenyl + H2O
-
Clostridium perfringens ?
-
?

Subunits

Subunits Comment Organism
? x * 88000, SDS-PAGE Clostridium perfringens

Synonyms

Synonyms Comment Organism
EABase
-
Clostridium perfringens
endo-beta-galactosidase
-
Clostridium perfringens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.17
-
2,4-dinitrophenyl 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-3)-[alpha-L-fucopyranosyl-(1-2)]-beta-D-galactopyranoside mutant enzyme E354A, in 20 mM sodium acetate buffer at pH 6.0, 37°C Clostridium perfringens
1.35
-
2,4-dinitrophenyl 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-3)-[alpha-L-fucopyranosyl-(1-2)]-beta-D-galactopyranoside mutant enzyme D429A, in 20 mM sodium acetate buffer at pH 6.0, 37°C Clostridium perfringens
1.55
-
2,4-dinitrophenyl 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-3)-[alpha-L-fucopyranosyl-(1-2)]-beta-D-galactopyranoside mutant enzyme E467A, in 20 mM sodium acetate buffer at pH 6.0, 37°C Clostridium perfringens
1.75
-
2,4-dinitrophenyl 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-3)-[alpha-L-fucopyranosyl-(1-2)]-beta-D-galactopyranoside wild type enzyme, in 20 mM sodium acetate buffer at pH 6.0, 37°C Clostridium perfringens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.33
-
2,4-dinitrophenyl 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-3)-[alpha-L-fucopyranosyl-(1-2)]-beta-D-galactopyranoside mutant enzyme E354A, in 20 mM sodium acetate buffer at pH 6.0, 37°C Clostridium perfringens
21.67
-
2,4-dinitrophenyl 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-3)-[alpha-L-fucopyranosyl-(1-2)]-beta-D-galactopyranoside mutant enzyme D429A, in 20 mM sodium acetate buffer at pH 6.0, 37°C Clostridium perfringens
26.67
-
2,4-dinitrophenyl 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-3)-[alpha-L-fucopyranosyl-(1-2)]-beta-D-galactopyranoside mutant enzyme E467A, in 20 mM sodium acetate buffer at pH 6.0, 37°C Clostridium perfringens
26.67
-
2,4-dinitrophenyl 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-3)-[alpha-L-fucopyranosyl-(1-2)]-beta-D-galactopyranoside wild type enzyme, in 20 mM sodium acetate buffer at pH 6.0, 37°C Clostridium perfringens