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Literature summary 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

Activating Compound

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

Cloned(Commentary)

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

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.7.43 4-(hydroxymercuri) benzoic acid
-
 Acetivibrio thermocellus
2.7.7.43 4-(hydroxymercuri)benzoic acid
-
 Acetivibrio thermocellus
2.7.7.43 Ca2+ 95% inhibition of enzyme activity at 10 mM Acetivibrio thermocellus
2.7.7.43 Cu2+ 97% inhibition of enzyme activity at 10 mM Acetivibrio thermocellus
2.7.7.43 Fe3+ 91% inhibition of enzyme activity at 10 mM Acetivibrio thermocellus
2.7.7.43 additional information the addition of monovalent cations (K+, Na+) to the reaction mixtures has no effect on the enzyme activity Acetivibrio thermocellus
2.7.7.43 N-ethylmaleimide
-
 Acetivibrio thermocellus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.43 0.13
-
 NeuAc
-
 Acetivibrio thermocellus
2.7.7.43 0.13
-
 N-acylneuraminate at 50°C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5) Acetivibrio thermocellus
2.7.7.43 0.16
-
 NeuGc
-
 Acetivibrio thermocellus
2.7.7.43 0.16
-
 N-glycolylneuraminic acid at 50°C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5) Acetivibrio thermocellus
2.7.7.43 0.24
-
 CTP
-
 Acetivibrio thermocellus
2.7.7.43 0.24
-
 CTP at 50°C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5) Acetivibrio thermocellus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.7.43 Co2+ 66% as effective as Mn2+ Acetivibrio thermocellus
2.7.7.43 Co2+ can replace Mn2+, 66% as effective as Mn2+ at 10 mM Acetivibrio thermocellus
2.7.7.43 Mg2+ 20 mM Acetivibrio thermocellus
2.7.7.43 Mg2+ 57% as effective as Mn2+ Acetivibrio thermocellus
2.7.7.43 Mg2+ can replace Mn2+, 57% as effective as Mn2+ at 10 mM Acetivibrio thermocellus
2.7.7.43 Mn2+ absolute requirement for a divalent cation for activity, maximum activity at 10 mM Mn2+ Acetivibrio thermocellus
2.7.7.43 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
2.7.7.43 Zn2+ 62% as effective as Mn2+ Acetivibrio thermocellus
2.7.7.43 Zn2+ can replace Mn2+, 62% as effective as Mn2+ at 10 mM Acetivibrio thermocellus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.7.43 26000
-
 SDS-PAGE Acetivibrio thermocellus

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.43 Acetivibrio thermocellus
-
 strain ATCC 27405
-
2.7.7.43 Acetivibrio thermocellus
-
 ATCC27405
-
2.7.7.43 Acetivibrio thermocellus ATCC 27405
-
 ATCC27405
-

Purification (Commentary)

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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.43 CTP + N-acylneuraminate
-
 Acetivibrio thermocellus diphosphate + CMP-N-acylneuraminate
-
 ?
2.7.7.43 CTP + N-acylneuraminate
-
 Acetivibrio thermocellus ATCC 27405 diphosphate + CMP-N-acylneuraminate
-
 ?
2.7.7.43 CTP + N-glycolylneuraminic acid
-
 Acetivibrio thermocellus CMP-N-glycolylneuraminate + diphosphate
-
 ?
2.7.7.43 CTP + N-glycolylneuraminic acid
-
 Acetivibrio thermocellus ATCC 27405 CMP-N-glycolylneuraminate + diphosphate
-
 ?
2.7.7.43 N-acetylneuraminic acid + CTP bi-bi catalytic mechanism, enzyme also accepts N-glycolylneuraminic acid (NeuGc) as a substrate Acetivibrio thermocellus CMP-NeuAc + diphosphate
-
 ?
2.7.7.43 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
-
 ?
2.7.7.43 N-glycolylneuraminic acid + CTP
-
 Acetivibrio thermocellus CMP-N-glycolylneuraminate + diphosphate
-
 ?
2.7.7.43 N-glycolylneuraminic acid + CTP
-
 Acetivibrio thermocellus ATCC 27405 CMP-N-glycolylneuraminate + diphosphate
-
 ?

Synonyms

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.7.43 50
-
-
 Acetivibrio thermocellus
2.7.7.43 50
-
 maximum activity Acetivibrio thermocellus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.7.7.43 37 60 enzyme is active between 37°C - 60°C Acetivibrio thermocellus
2.7.7.43 37 60 in the presence of 0.2 mM dithiothreitol Acetivibrio thermocellus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.7.7.43 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]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.7.43 1.66
-
 NeuGc
-
 Acetivibrio thermocellus
2.7.7.43 1.66
-
 N-glycolylneuraminic acid at 50°C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5) Acetivibrio thermocellus
2.7.7.43 2.25
-
 NeuAc
-
 Acetivibrio thermocellus
2.7.7.43 2.25
-
 N-acylneuraminate at 50°C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5) Acetivibrio thermocellus
2.7.7.43 3.33
-
 CTP
-
 Acetivibrio thermocellus
2.7.7.43 3.33
-
 CTP at 50°C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5) Acetivibrio thermocellus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.7.43 9.5
-
-
 Acetivibrio thermocellus
2.7.7.43 9.5
-
 maximum activity Acetivibrio thermocellus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.7.7.43 8 13 enzyme is active between pH 8-13 Acetivibrio thermocellus
2.7.7.43 8 13 in Tris-HCl buffer Acetivibrio thermocellus