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Literature summary for 4.1.3.3 extracted from

  • Kruger, D.; Schauer, R.; Traving, C.
    Characterization and mutagenesis of the recombinant N-acetylneuraminate lyase from Clostridium perfringens: insights into the reaction mechanism (2001), Eur. J. Biochem., 268, 3831-3839.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Clostridium perfringens

Protein Variants

Protein Variants Comment Organism
D187E increased Km Clostridium perfringens
D187N decreased Vmax, increased Km Clostridium perfringens
E188D cleaves 5-N-acetyl-9-O-acetylneuraminic acid as good as N-acetylneuraminic acid Clostridium perfringens
E188Q cleaves 5-N-acetyl-9-O-acetylneuraminic acid as good as N-acetylneuraminic acid Clostridium perfringens
K161R decreased Vmax Clostridium perfringens
Y133C significant decrease in enzyme activity Clostridium perfringens
Y133F significant decrease in enzyme activity Clostridium perfringens
Y133H significant decrease in enzyme activity Clostridium perfringens
Y133W significant decrease in enzyme activity Clostridium perfringens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.4
-
N-acetylneuraminic acid pH 7.2, Y133F mutant Clostridium perfringens
1
-
N-acetylneuraminic acid pH 7.2, Y133H mutant Clostridium perfringens
2
-
N-acetylneuraminic acid pH 7.2, 50 mM phosphate buffer, wild-type Clostridium perfringens
2.4
-
N-acetylneuraminic acid pH 7.2, Y133C mutant Clostridium perfringens
2.9
-
N-acetylneuraminic acid pH 7.2, Y133W mutant Clostridium perfringens
3.2
-
N-acetylneuraminic acid pH 7.2, 250 mM phosphate buffer, wild-type Clostridium perfringens
3.5
-
N-acetylneuraminic acid pH 7.2, K161R mutant Clostridium perfringens
17
-
N-acetylneuraminic acid pH 7.2, E188Q mutant Clostridium perfringens
29
-
N-acetylneuraminic acid pH 7.2, D187E mutant Clostridium perfringens
44
-
N-acetylneuraminic acid pH 7.2, E188D mutant Clostridium perfringens
1100
-
N-acetylneuraminic acid pH 7.2, D187N mutant Clostridium perfringens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
34600
-
SDS-PAGE Clostridium perfringens

Organism

Organism UniProt Comment Textmining
Clostridium perfringens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
homogeneity Clostridium perfringens

Reaction

Reaction Comment Organism Reaction ID
aceneuramate = N-acetyl-D-mannosamine + pyruvate mechanism Clostridium perfringens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
11.36
-
-
Clostridium perfringens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5-N-acetyl-9-O-acetylneuraminic acid 54% the activity with N-acetylneuraminic acid Clostridium perfringens ?
-
?
N-Acetylneuraminic acid
-
Clostridium perfringens N-Acetyl-D-mannosamine + pyruvate
-
r

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
65.7
-
-
Clostridium perfringens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
60
-
increased activity after 10 min Clostridium perfringens
70
-
no loss of activity after 10 min Clostridium perfringens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
-
Clostridium perfringens