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Literature summary extracted from

  • Garcia-Garcia, M.I.; Gil-Ortiz, F.; Garcia-Carmona, F.; Sanchez-Ferrer, A.
    First functional and mutational analysis of group 3 N-acetylneuraminate lyases from Lactobacillus antri and Lactobacillus sakei 23K (2014), PLoS ONE, 9, e96976 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.3.3 expressed in Escherichia coli Rosetta 2 cells Latilactobacillus sakei
4.1.3.3 expressed in Escherichia coli Rosetta 2 cells Limosilactobacillus antri

Protein Variants

EC Number Protein Variants Comment Organism
4.1.3.3 G210S/Y213G the mutant shows reduced activity compared to the wild type enzyme Limosilactobacillus antri
4.1.3.3 G211S the mutant shows reduced activity compared to the wild type enzyme Limosilactobacillus antri
4.1.3.3 P193Y the mutant shows reduced activity compared to the wild type enzyme Limosilactobacillus antri

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.1.3.3 0.27
-
N-acetylneuraminate mutant enzyme P193Y, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 0.32
-
N-acetylneuraminate wild type enzyme, at pH 7.0 and 37°C Latilactobacillus sakei
4.1.3.3 0.5
-
pyruvate mutant enzyme P193Y, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 1.1
-
pyruvate wild type enzyme, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 1.1
-
N-acetylneuraminate wild type enzyme, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 1.2
-
N-acetylneuraminate mutant enzyme G211S, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 2
-
N-acetylneuraminate mutant enzyme G210S/Y213G, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 4.3
-
pyruvate mutant enzyme G210S/Y213G, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 12.5
-
pyruvate wild type enzyme, at pH 7.0 and 37°C Latilactobacillus sakei
4.1.3.3 272
-
N-acetyl-D-mannosamine wild type enzyme, at pH 7.0 and 37°C Latilactobacillus sakei
4.1.3.3 333
-
N-acetyl-D-mannosamine wild type enzyme, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 915
-
N-acetyl-D-mannosamine mutant enzyme P193Y, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 3200
-
N-acetyl-D-mannosamine mutant enzyme G210S/Y213G, at pH 7.0 and 37°C Limosilactobacillus antri

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.1.3.3 140000
-
gel filtration Latilactobacillus sakei
4.1.3.3 140000
-
gel filtration Limosilactobacillus antri

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.1.3.3 N-acetylneuraminate Latilactobacillus sakei 100% activity N-acetyl-D-mannosamine + pyruvate
-
r
4.1.3.3 N-acetylneuraminate Limosilactobacillus antri 100% activity N-acetyl-D-mannosamine + pyruvate
-
r
4.1.3.3 N-acetylneuraminate Latilactobacillus sakei 23K 100% activity N-acetyl-D-mannosamine + pyruvate
-
r
4.1.3.3 N-acetylneuraminate Limosilactobacillus antri DSMZ 16041 100% activity N-acetyl-D-mannosamine + pyruvate
-
r

Organism

EC Number Organism UniProt Comment Textmining
4.1.3.3 Latilactobacillus sakei
-
-
-
4.1.3.3 Latilactobacillus sakei 23K
-
-
-
4.1.3.3 Limosilactobacillus antri C8P490
-
-
4.1.3.3 Limosilactobacillus antri DSMZ 16041 C8P490
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.3.3 Ni2+-chelating affinity column chromatography Latilactobacillus sakei
4.1.3.3 Ni2+-chelating affinity column chromatography Limosilactobacillus antri

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.3.3 2-deoxy-D-glucose + pyruvate 1.2% activity compared to N-acetylneuraminate Limosilactobacillus antri ?
-
r
4.1.3.3 2-deoxy-D-glucose + pyruvate 33% activity compared to N-acetylneuraminate Latilactobacillus sakei ?
-
r
4.1.3.3 2-deoxy-D-glucose + pyruvate 33% activity compared to N-acetylneuraminate Latilactobacillus sakei 23K ?
-
r
4.1.3.3 2-deoxy-D-glucose + pyruvate 1.2% activity compared to N-acetylneuraminate Limosilactobacillus antri DSMZ 16041 ?
-
r
4.1.3.3 D-arabinose + pyruvate 1.0% activity compared to N-acetylneuraminate Latilactobacillus sakei 3-deoxy-D-manno-2-octulosonic acid
-
r
4.1.3.3 D-arabinose + pyruvate 1.5% activity compared to N-acetylneuraminate Limosilactobacillus antri 3-deoxy-D-manno-2-octulosonic acid
-
r
4.1.3.3 D-mannose + pyruvate 10% activity compared to N-acetylneuraminate Limosilactobacillus antri ?
-
r
4.1.3.3 D-mannose + pyruvate 95% activity compared to N-acetylneuraminate Latilactobacillus sakei ?
-
r
4.1.3.3 D-mannose + pyruvate 95% activity compared to N-acetylneuraminate Latilactobacillus sakei 23K ?
-
r
4.1.3.3 D-mannose + pyruvate 10% activity compared to N-acetylneuraminate Limosilactobacillus antri DSMZ 16041 ?
-
r
4.1.3.3 D-xylose + pyruvate 0.6% activity compared to N-acetylneuraminate Limosilactobacillus antri ?
-
r
4.1.3.3 D-xylose + pyruvate 2.4% activity compared to N-acetylneuraminate Latilactobacillus sakei ?
-
r
4.1.3.3 D-xylose + pyruvate 2.4% activity compared to N-acetylneuraminate Latilactobacillus sakei 23K ?
-
r
4.1.3.3 D-xylose + pyruvate 0.6% activity compared to N-acetylneuraminate Limosilactobacillus antri DSMZ 16041 ?
-
r
4.1.3.3 L-gulose + pyruvate 0.7% activity compared to N-acetylneuraminate Limosilactobacillus antri ?
-
r
4.1.3.3 L-gulose + pyruvate 3.6% activity compared to N-acetylneuraminate Latilactobacillus sakei ?
-
r
4.1.3.3 L-gulose + pyruvate 3.6% activity compared to N-acetylneuraminate Latilactobacillus sakei 23K ?
-
r
4.1.3.3 L-gulose + pyruvate 0.7% activity compared to N-acetylneuraminate Limosilactobacillus antri DSMZ 16041 ?
-
r
4.1.3.3 N-acetylneuraminate 100% activity Latilactobacillus sakei N-acetyl-D-mannosamine + pyruvate
-
r
4.1.3.3 N-acetylneuraminate 100% activity Limosilactobacillus antri N-acetyl-D-mannosamine + pyruvate
-
r
4.1.3.3 N-acetylneuraminate 100% activity Latilactobacillus sakei 23K N-acetyl-D-mannosamine + pyruvate
-
r
4.1.3.3 N-acetylneuraminate 100% activity Limosilactobacillus antri DSMZ 16041 N-acetyl-D-mannosamine + pyruvate
-
r

Subunits

EC Number Subunits Comment Organism
4.1.3.3 homotetramer 4 * 35000, SDS-PAGE Latilactobacillus sakei
4.1.3.3 homotetramer 4 * 35000, SDS-PAGE Limosilactobacillus antri
4.1.3.3 homotetramer 4 * 33500, electrospray ionization mass spectrometry Latilactobacillus sakei
4.1.3.3 homotetramer 4 * 35700, electrospray ionization mass spectrometry Limosilactobacillus antri

Synonyms

EC Number Synonyms Comment Organism
4.1.3.3 NAL
-
Latilactobacillus sakei
4.1.3.3 NAL
-
Limosilactobacillus antri
4.1.3.3 Neu5Ac aldolase
-
Latilactobacillus sakei
4.1.3.3 Neu5Ac aldolase
-
Limosilactobacillus antri

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.1.3.3 40
-
in synthetic reaction Latilactobacillus sakei
4.1.3.3 50
-
in synthetic reaction Latilactobacillus sakei
4.1.3.3 60
-
in synthetic and hydrolytic reactions Limosilactobacillus antri

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4.1.3.3 42
-
melting temperature in 100 mM phosphate buffer pH 7.0 Latilactobacillus sakei
4.1.3.3 65
-
melting temperature in 100 mM phosphate buffer pH 7.0 Limosilactobacillus antri

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.1.3.3 0.032
-
N-acetylneuraminate mutant enzyme G211S, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 0.13
-
N-acetylneuraminate mutant enzyme P193Y, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 0.47
-
N-acetylneuraminate mutant enzyme G210S/Y213G, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 0.9
-
N-acetylneuraminate wild type enzyme, at pH 7.0 and 37°C Latilactobacillus sakei
4.1.3.3 0.99
-
pyruvate mutant enzyme P193Y, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 1.9
-
N-acetylneuraminate wild type enzyme, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 2.4
-
N-acetyl-D-mannosamine mutant enzyme P193Y, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 3
-
pyruvate mutant enzyme G210S/Y213G, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 3.9
-
pyruvate wild type enzyme, at pH 7.0 and 37°C Latilactobacillus sakei
4.1.3.3 4.6
-
N-acetyl-D-mannosamine wild type enzyme, at pH 7.0 and 37°C Latilactobacillus sakei
4.1.3.3 5.4
-
N-acetyl-D-mannosamine wild type enzyme, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 5.6
-
pyruvate wild type enzyme, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 15.3
-
N-acetyl-D-mannosamine mutant enzyme G210S/Y213G, at pH 7.0 and 37°C Limosilactobacillus antri

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.1.3.3 6 7
-
Latilactobacillus sakei
4.1.3.3 6 7
-
Limosilactobacillus antri

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.1.3.3 0.002
-
N-acetyl-D-mannosamine mutant enzyme P193Y, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 0.004
-
N-acetyl-D-mannosamine mutant enzyme G210S/Y213G, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 0.016
-
N-acetyl-D-mannosamine wild type enzyme, at pH 7.0 and 37°C Latilactobacillus sakei
4.1.3.3 0.016
-
N-acetyl-D-mannosamine wild type enzyme, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 0.026
-
N-acetylneuraminate mutant enzyme G211S, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 0.23
-
N-acetylneuraminate mutant enzyme G210S/Y213G, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 0.31
-
pyruvate wild type enzyme, at pH 7.0 and 37°C Latilactobacillus sakei
4.1.3.3 0.48
-
N-acetylneuraminate mutant enzyme P193Y, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 0.71
-
pyruvate mutant enzyme G210S/Y213G, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 1.7
-
N-acetylneuraminate wild type enzyme, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 1.98
-
pyruvate mutant enzyme P193Y, at pH 7.0 and 37°C Limosilactobacillus antri
4.1.3.3 2.8
-
N-acetylneuraminate wild type enzyme, at pH 7.0 and 37°C Latilactobacillus sakei
4.1.3.3 5.1
-
pyruvate wild type enzyme, at pH 7.0 and 37°C Limosilactobacillus antri