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Literature summary for 4.1.3.3 extracted from

  • Ji, W.; Sun, W.; Feng, J.; Song, T.; Zhang, D.; Ouyang, P.; Gu, Z.; Xie, J.
    Characterization of a novel N-acetylneuraminic acid lyase favoring industrial N-acetylneuraminic acid synthesis (2015), Sci. Rep., 5, 9341 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 4.1.3.3

Activating Compound

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(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rosetta (DE3) cells Corynebacterium glutamicum

Inhibitors

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 [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

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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Corynebacterium glutamicum Q8NMD2
-
-
Corynebacterium glutamicum ATCC 13032 Q8NMD2
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA agarose column chromatography Corynebacterium glutamicum

Substrates and Products (Substrate)

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

Subunits Comment Organism
? x * 33000, SDS-PAGE Corynebacterium glutamicum

Synonyms

Synonyms Comment Organism
N-Acetylneuraminic acid lyase
-
 Corynebacterium glutamicum
NAL
-
 Corynebacterium glutamicum

Temperature Optimum [°C]

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 [°C]

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 [1/s]

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

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 [mM/s]

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