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Literature summary for 2.7.7.43 extracted from

  • Mizanur, R.M.; Pohl, N.L.
    Cloning and characterization of a heat-stable CMP-N-acylneuraminic acid synthetase from Clostridium thermocellum (2007), Appl. Microbiol. Biotechnol., 76, 827-834.
    View publication on PubMed
Search for more literature for 2.7.7.43

Activating Compound

Activating Compound Comment Organism Structure
dithiothreitol Dithiothreitol at a concentration of 0.2 mM stimulates enzyme activity by 158% Acetivibrio thermocellus
DTT maximum activity in the presence of 0.2 mM DTT Acetivibrio thermocellus
additional information mercaptoethanol has no stimulatory effect Acetivibrio thermocellus

Cloned(Commentary)

Cloned (Comment) Organism
cloned in Escherichia coli XL-10 Acetivibrio thermocellus
expressed in Escherichia coli strain BL21 Acetivibrio thermocellus

Inhibitors

Inhibitors Comment Organism Structure
4-(hydroxymercuri) benzoic acid
-
 Acetivibrio thermocellus
4-(hydroxymercuri)benzoic acid
-
 Acetivibrio thermocellus
Ca2+ 95% inhibition of enzyme activity at 10 mM Acetivibrio thermocellus
Cu2+ 97% inhibition of enzyme activity at 10 mM Acetivibrio thermocellus
Fe3+ 91% inhibition of enzyme activity at 10 mM Acetivibrio thermocellus
additional information the addition of monovalent cations (K+, Na+) to the reaction mixtures has no effect on the enzyme activity Acetivibrio thermocellus
N-ethylmaleimide
-
 Acetivibrio thermocellus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.13
-
 NeuAc
-
 Acetivibrio thermocellus
0.13
-
 N-acylneuraminate at 50°C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5) Acetivibrio thermocellus
0.16
-
 NeuGc
-
 Acetivibrio thermocellus
0.16
-
 N-glycolylneuraminic acid at 50°C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5) Acetivibrio thermocellus
0.24
-
 CTP
-
 Acetivibrio thermocellus
0.24
-
 CTP at 50°C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5) Acetivibrio thermocellus

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ 66% as effective as Mn2+ Acetivibrio thermocellus
Co2+ can replace Mn2+, 66% as effective as Mn2+ at 10 mM Acetivibrio thermocellus
Mg2+ 20 mM Acetivibrio thermocellus
Mg2+ 57% as effective as Mn2+ Acetivibrio thermocellus
Mg2+ can replace Mn2+, 57% as effective as Mn2+ at 10 mM Acetivibrio thermocellus
Mn2+ absolute requirement for a divalent cation for activity, maximum activity at 10 mM Mn2+ Acetivibrio thermocellus
Mn2+ the enzyme has an absolute requirement for a divalent cation for activity and reaches maximum activity in the presence of 10 mM Mn2+ Acetivibrio thermocellus
Zn2+ 62% as effective as Mn2+ Acetivibrio thermocellus
Zn2+ can replace Mn2+, 62% as effective as Mn2+ at 10 mM Acetivibrio thermocellus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
26000
-
 SDS-PAGE Acetivibrio thermocellus

Organism

Organism UniProt Comment Textmining
Acetivibrio thermocellus
-
 strain ATCC 27405
-
Acetivibrio thermocellus
-
 ATCC27405
-
Acetivibrio thermocellus ATCC 27405
-
 ATCC27405
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Acetivibrio thermocellus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
CTP + N-acylneuraminate
-
 Acetivibrio thermocellus diphosphate + CMP-N-acylneuraminate
-
 ?
CTP + N-acylneuraminate
-
 Acetivibrio thermocellus ATCC 27405 diphosphate + CMP-N-acylneuraminate
-
 ?
CTP + N-glycolylneuraminic acid
-
 Acetivibrio thermocellus CMP-N-glycolylneuraminate + diphosphate
-
 ?
CTP + N-glycolylneuraminic acid
-
 Acetivibrio thermocellus ATCC 27405 CMP-N-glycolylneuraminate + diphosphate
-
 ?
N-acetylneuraminic acid + CTP bi-bi catalytic mechanism, enzyme also accepts N-glycolylneuraminic acid (NeuGc) as a substrate Acetivibrio thermocellus CMP-NeuAc + diphosphate
-
 ?
N-acetylneuraminic acid + CTP bi-bi catalytic mechanism, enzyme also accepts N-glycolylneuraminic acid (NeuGc) as a substrate Acetivibrio thermocellus ATCC 27405 CMP-NeuAc + diphosphate
-
 ?
N-glycolylneuraminic acid + CTP
-
 Acetivibrio thermocellus CMP-N-glycolylneuraminate + diphosphate
-
 ?
N-glycolylneuraminic acid + CTP
-
 Acetivibrio thermocellus ATCC 27405 CMP-N-glycolylneuraminate + diphosphate
-
 ?

Synonyms

Synonyms Comment Organism
CMP-N-acylneuraminic acid synthetase
-
 Acetivibrio thermocellus
CMP-NeuAc synthetase
-
 Acetivibrio thermocellus
CSS
-
 Acetivibrio thermocellus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
-
 Acetivibrio thermocellus
50
-
 maximum activity Acetivibrio thermocellus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
37 60 enzyme is active between 37°C - 60°C Acetivibrio thermocellus
37 60 in the presence of 0.2 mM dithiothreitol Acetivibrio thermocellus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
37 70 complete loss of activity above 70°C, no loss of activity after 24 h incubation at 37°C and 50°C Acetivibrio thermocellus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.66
-
 NeuGc
-
 Acetivibrio thermocellus
1.66
-
 N-glycolylneuraminic acid at 50°C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5) Acetivibrio thermocellus
2.25
-
 NeuAc
-
 Acetivibrio thermocellus
2.25
-
 N-acylneuraminate at 50°C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5) Acetivibrio thermocellus
3.33
-
 CTP
-
 Acetivibrio thermocellus
3.33
-
 CTP at 50°C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5) Acetivibrio thermocellus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9.5
-
-
 Acetivibrio thermocellus
9.5
-
 maximum activity Acetivibrio thermocellus

pH Range

pH Minimum pH Maximum Comment Organism
8 13 enzyme is active between pH 8-13 Acetivibrio thermocellus
8 13 in Tris-HCl buffer Acetivibrio thermocellus