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Literature summary for 4.1.3.3 extracted from

  • Garcia Garcia, M.I.; Sola Carvajal, A.; Garcia Carmona, F.; Sanchez Ferrer, A.
    Characterization of a novel N-acetylneuraminate lyase from Staphylococcus carnosus TM300 and its application to N-acetylneuraminic acid production (2012), J. Agric. Food Chem., 60, 7450-7456.
    View publication on PubMed

Application

Application Comment Organism
synthesis the enzyme can be a useful and cheap biocatalyst to produce N-acetylneuraminic acid (Neu5Ac) Staphylococcus carnosus

Cloned(Commentary)

Cloned (Comment) Organism
gene ScNAL, sequence comparison, expression of His-tagged enzyme in Escherichia coli strain Rosetta (DE3) pLys Staphylococcus carnosus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1 4 pyruvate recombinant His-taggged enzyme, pH 7.0, 37°C Staphylococcus carnosus
2
-
N-acetylneuraminate recombinant His-taggged enzyme, pH 7.0, 37°C Staphylococcus carnosus
149
-
N-acetyl-D-mannosamine recombinant His-taggged enzyme, pH 7.0, 37°C Staphylococcus carnosus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
N-acetyl-D-mannosamine + pyruvate Staphylococcus carnosus reversible aldol condensation of pyruvate and N-acetyl-D-mannosamine N-acetylneuraminate
-
r
N-acetyl-D-mannosamine + pyruvate Staphylococcus carnosus TM300 reversible aldol condensation of pyruvate and N-acetyl-D-mannosamine N-acetylneuraminate
-
r
N-acetylneuraminate Staphylococcus carnosus
-
N-acetyl-D-mannosamine + pyruvate
-
r
N-acetylneuraminate Staphylococcus carnosus TM300
-
N-acetyl-D-mannosamine + pyruvate
-
r

Organism

Organism UniProt Comment Textmining
Staphylococcus carnosus B9DIJ2
-
-
Staphylococcus carnosus TM300 B9DIJ2
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme 7.5fold from Escherichia coli strain Rosetta (DE3) pLys by ultrafiltration, nickel affinity chromatography, and gel filtration, to homogeneity Staphylococcus carnosus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
12.7
-
purified recombinant His-taggged enzyme, pH 7.0, 37°C Staphylococcus carnosus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
N-acetyl-D-mannosamine + pyruvate reversible aldol condensation of pyruvate and N-acetyl-D-mannosamine Staphylococcus carnosus N-acetylneuraminate
-
r
N-acetyl-D-mannosamine + pyruvate reversible aldol condensation of pyruvate and N-acetyl-D-mannosamine Staphylococcus carnosus TM300 N-acetylneuraminate
-
r
N-acetylneuraminate
-
Staphylococcus carnosus N-acetyl-D-mannosamine + pyruvate
-
r
N-acetylneuraminate
-
Staphylococcus carnosus TM300 N-acetyl-D-mannosamine + pyruvate
-
r

Synonyms

Synonyms Comment Organism
NAL
-
Staphylococcus carnosus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
synthesis reaction Staphylococcus carnosus
60
-
hydrolysis reaction Staphylococcus carnosus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
20 80 synthesis and hydrolysis reactions, profiles, overview Staphylococcus carnosus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
20 50 purified recombinant His-tagged enzyme, pH 7.0, completely stable over 140-160 min Staphylococcus carnosus
60
-
purified recombinant His-tagged enzyme, pH 7.0, 120 min, 60% activity remaining Staphylococcus carnosus
70
-
purified recombinant His-tagged enzyme, pH 7.0, inactivation within 60 min Staphylococcus carnosus
80
-
purified recombinant His-tagged enzyme, pH 7.0, inactivation within 10 min Staphylococcus carnosus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.9
-
pyruvate recombinant His-taggged enzyme, pH 7.0, 37°C Staphylococcus carnosus
4
-
N-acetylneuraminate recombinant His-taggged enzyme, pH 7.0, 37°C Staphylococcus carnosus
4.5
-
N-acetyl-D-mannosamine recombinant His-taggged enzyme, pH 7.0, 37°C Staphylococcus carnosus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Staphylococcus carnosus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.03
-
N-acetyl-D-mannosamine recombinant His-taggged enzyme, pH 7.0, 37°C Staphylococcus carnosus
0.21
-
pyruvate recombinant His-taggged enzyme, pH 7.0, 37°C Staphylococcus carnosus
2
-
N-acetylneuraminate recombinant His-taggged enzyme, pH 7.0, 37°C Staphylococcus carnosus