Activating Compound | Comment | Organism | Structure |
---|---|---|---|
EDTA | about 120% activity at 5 mM and pH 7.5 for N-acetylneuraminate synthesis and about 150% activity at 5 mM and pH 8.5 for N-acetylneuraminate cleavage | Corynebacterium glutamicum | |
Triton X-100 | about 150% activity at 5 mM and pH 8.5 for N-acetylneuraminate synthesis and about 220% activity at 5 mM and pH 8.5 for N-acetylneuraminate cleavage | Corynebacterium glutamicum |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli Rosetta (DE3) cells | Corynebacterium glutamicum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
CTAB | about 30% residual activity at 5 mM and pH 7.5 for N-acetylneuraminate synthesis and almost complete inhibition at 5 mM for N-acetylneuraminate cleavage | Corynebacterium glutamicum | |
SDS | about 30% residual activity at 5 mM and pH 7.5 for N-acetylneuraminate synthesis and almost complete inhibition at 5 mM for N-acetylneuraminate cleavage | Corynebacterium glutamicum | |
Zn2+ | about 25% residual activity at 5 mM and pH 7.5 for N-acetylneuraminate synthesis and almost complete inhibition at 5 mM and pH 7.5 for N-acetylneuraminate cleavage | Corynebacterium glutamicum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3 | 3.5 | N-acetylneuraminate | at pH 7.5 and 37°C | Corynebacterium glutamicum | |
14.7 | - |
pyruvate | at pH 7.5 and 37°C | Corynebacterium glutamicum | |
53.3 | - |
N-acetyl-D-mannosamine | at pH 7.5 and 37°C | Corynebacterium glutamicum | |
72.4 | - |
pyruvate | at pH 8.5 and 37°C | Corynebacterium glutamicum | |
87.7 | - |
N-acetylneuraminate | at pH 8.5 and 37°C | Corynebacterium glutamicum | |
92.1 | - |
N-acetyl-D-mannosamine | at pH 8.5 and 37°C | Corynebacterium glutamicum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ba2+ | about 170% activity at 5 mM and pH 7.5 for N-acetylneuraminate synthesis | Corynebacterium glutamicum | |
Co2+ | about 225% activity at 5 mM and pH 7.5 for N-acetylneuraminate synthesis | Corynebacterium glutamicum | |
Fe3+ | about 125% activity at 5 mM and pH 7.5 for N-acetylneuraminate synthesis | Corynebacterium glutamicum | |
Mg2+ | about 270% activity at 5 mM and pH 7.5 for N-acetylneuraminate synthesis | Corynebacterium glutamicum | |
Ni2+ | about 220% activity at 5 mM and pH 7.5 for N-acetylneuraminate synthesis | Corynebacterium glutamicum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
N-acetylneuraminate | Corynebacterium glutamicum | - |
N-acetyl-D-mannosamine + pyruvate | - |
r | |
N-acetylneuraminate | Corynebacterium glutamicum ATCC 13032 | - |
N-acetyl-D-mannosamine + pyruvate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Corynebacterium glutamicum | Q8NMD2 | - |
- |
Corynebacterium glutamicum ATCC 13032 | Q8NMD2 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA agarose column chromatography | Corynebacterium glutamicum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
N-acetyl-D-mannosamine + pyruvate | - |
Corynebacterium glutamicum | N-acetylneuraminate | - |
r | |
N-acetyl-D-mannosamine + pyruvate | - |
Corynebacterium glutamicum ATCC 13032 | N-acetylneuraminate | - |
r | |
N-acetylneuraminate | - |
Corynebacterium glutamicum | N-acetyl-D-mannosamine + pyruvate | - |
r | |
N-acetylneuraminate | - |
Corynebacterium glutamicum ATCC 13032 | N-acetyl-D-mannosamine + pyruvate | - |
r |
Subunits | Comment | Organism |
---|---|---|
? | x * 33000, SDS-PAGE | Corynebacterium glutamicum |
Synonyms | Comment | Organism |
---|---|---|
N-Acetylneuraminic acid lyase | - |
Corynebacterium glutamicum |
NAL | - |
Corynebacterium glutamicum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
for N-acetylneuraminate cleavage at pH 7.5 and N-acetylneuraminate synthesis at pH 8.5 | Corynebacterium glutamicum |
40 | 50 | for N-acetylneuraminate synthesis at pH 7.5 | Corynebacterium glutamicum |
45 | - |
for N-acetylneuraminate cleavage at pH 8.5 | Corynebacterium glutamicum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
when the enzyme is incubated at 40°C, pH 8.5 for 48 h, in N-acetylneuraminate synthesis direction, the enzyme keeps 100% activity for the first 24 h but remains 60% of its activity in 48h. 75% and 20% of N-acetylneuraminate cleavage activity remain in 24 h and 48 h, respectively | Corynebacterium glutamicum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5.67 | - |
N-acetyl-D-mannosamine | at pH 7.5 and 37°C | Corynebacterium glutamicum | |
6.1 | - |
pyruvate | at pH 7.5 and 37°C | Corynebacterium glutamicum | |
9.3 | - |
N-acetylneuraminate | at pH 7.5 and 37°C | Corynebacterium glutamicum | |
40.67 | - |
N-acetyl-D-mannosamine | at pH 8.5 and 37°C | Corynebacterium glutamicum | |
42.58 | - |
pyruvate | at pH 8.5 and 37°C | Corynebacterium glutamicum | |
44.2 | - |
N-acetylneuraminate | at pH 8.5 and 37°C | Corynebacterium glutamicum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.2 | 8.4 | for N-acetylneuraminate cleavage | Corynebacterium glutamicum |
8.6 | - |
for N-acetylneuraminate synthesis | Corynebacterium glutamicum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.07 | - |
N-acetyl-D-mannosamine | at pH 8.5 and 37°C | Corynebacterium glutamicum | |
0.11 | - |
N-acetyl-D-mannosamine | at pH 7.5 and 37°C | Corynebacterium glutamicum | |
0.11 | - |
pyruvate | at pH 8.5 and 37°C | Corynebacterium glutamicum | |
0.28 | - |
N-acetylneuraminate | at pH 7.5 and 37°C | Corynebacterium glutamicum | |
0.41 | - |
pyruvate | at pH 7.5 and 37°C | Corynebacterium glutamicum | |
23.75 | - |
N-acetylneuraminate | at pH 8.5 and 37°C | Corynebacterium glutamicum |