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Literature summary for 5.1.3.8 extracted from

  • Klermund, L.; Groher, A.; Castiglione, K.
    New N-acyl-D-glucosamine 2-epimerases from cyanobacteria with high activity in the absence of ATP and low inhibition by pyruvate (2013), J. Biotechnol., 168, 256-263.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
ATP allosteric activation Acaryochloris marina
ATP allosteric activation, ATP has a more pronounced effect on the Km for the substrate than on the enzyme activity. The enzyme shows 20% of maximal activity without ATP. At high substrate concentrations of 200 mM and without ATP, the enzyme reaches up to 32% of the activity measured with ATP in excess Trichormus variabilis
ATP allosteric activation, the enzyme shows 13% of maximal activity without ATP Nostoc sp.
ATP allosteric activation, the enzyme shows 18% of maximal activity without ATP Nostoc punctiforme

Cloned(Commentary)

Cloned (Comment) Organism
expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3) in inclusion bodies, subcloning in Escherichia coli strain DH5alpha Acaryochloris marina
expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha Trichormus variabilis
expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha Nostoc sp.
expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha Nostoc punctiforme

Inhibitors

Inhibitors Comment Organism Structure
N-acetylneuraminic acid linear competitive with respect to GlcNAc, kinetics of the reversible uni-uni reaction, overview Nostoc punctiforme
N-acetylneuraminic acid linear competitive with respect to GlcNAc, kinetics of the reversible uni-uni reaction, overview Nostoc sp.
N-acetylneuraminic acid linear competitive with respect to GlcNAc, kinetics of the reversible uni-uni reaction, overview Trichormus variabilis
pyruvate
-
Acaryochloris marina
pyruvate linear competitive with respect to GlcNAc, kinetics of the reversible uni-uni reaction, overview Nostoc punctiforme
pyruvate linear competitive with respect to GlcNAc, kinetics of the reversible uni-uni reaction, overview Nostoc sp.
pyruvate low inhibition, linear competitive with respect to GlcNAc, kinetics of the reversible uni-uni reaction, overview Trichormus variabilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2 2.2 N-acetyl-D-glucosamine pH 9.0, 30°C Trichormus variabilis
16.3
-
N-acetyl-D-glucosamine pH 7.5, 30°C Trichormus variabilis
16.6
-
N-acetyl-D-glucosamine pH 7.5, 30°C Nostoc punctiforme
16.6
-
N-acetyl-D-mannosamine pH 7.5, 30°C Nostoc punctiforme
16.7
-
N-acetyl-D-mannosamine pH 7.5, 30°C Trichormus variabilis
18.8
-
N-acetyl-D-glucosamine pH 7.5, 37°C Trichormus variabilis
18.8
-
N-acetyl-D-glucosamine pH 8.0, 30°C Trichormus variabilis
19.7
-
N-acetyl-D-mannosamine pH 7.5, 30°C Nostoc sp.
24.6
-
N-acetyl-D-glucosamine pH 7.5, 30°C Nostoc sp.
26.1
-
N-acetyl-D-glucosamine pH 7.0, 30°C Trichormus variabilis
62.5
-
N-acetyl-D-glucosamine pH 6.0, 30°C Trichormus variabilis
113.9
-
N-acetyl-D-glucosamine pH 7.5, 30°C, without ATP Trichormus variabilis
231.9
-
N-acetyl-D-glucosamine pH 7.5, 37°C, without ATP Trichormus variabilis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
N-acetyl-D-glucosamine Trichormus variabilis
-
N-acetyl-D-mannosamine
-
r
N-acetyl-D-glucosamine Nostoc sp.
-
N-acetyl-D-mannosamine
-
r
N-acetyl-D-glucosamine Nostoc punctiforme
-
N-acetyl-D-mannosamine
-
r
N-acetyl-D-glucosamine Acaryochloris marina
-
N-acetyl-D-mannosamine
-
r
N-acetyl-D-glucosamine Nostoc sp. PCC 7120
-
N-acetyl-D-mannosamine
-
r
N-acetyl-D-glucosamine Acaryochloris marina MBIC 11017
-
N-acetyl-D-mannosamine
-
r

Organism

Organism UniProt Comment Textmining
Acaryochloris marina
-
-
-
Acaryochloris marina MBIC 11017
-
-
-
Nostoc punctiforme
-
-
-
Nostoc sp.
-
-
-
Nostoc sp. PCC 7120
-
-
-
Trichormus variabilis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis Trichormus variabilis
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis Nostoc sp.
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis Nostoc punctiforme

Renatured (Commentary)

Renatured (Comment) Organism
recombinant His6-tagged enzyme expressed in Escherichia coli strain BL21(DE3) cannot be solubilized/purified from inclusion bodies Acaryochloris marina

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
79.2
-
with N-acetyl-D-glucosamine, pH 6.0, 30°C Trichormus variabilis
82.1
-
with N-acetyl-D-glucosamine, pH 7.0, 30°C Trichormus variabilis
82.4
-
with 250 mM N-acetyl-D-glucosamine, pH 9.0, 30°C Trichormus variabilis
90.8
-
with N-acetyl-D-glucosamine, pH 8.0, 30°C Trichormus variabilis
117
-
with N-acetyl-D-glucosamine, pH 7.5, 30°C Trichormus variabilis
131
-
with 430 mM N-acetyl-D-glucosamine, pH 9.0, 30°C Trichormus variabilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
N-acetyl-D-glucosamine
-
Trichormus variabilis N-acetyl-D-mannosamine
-
r
N-acetyl-D-glucosamine
-
Nostoc sp. N-acetyl-D-mannosamine
-
r
N-acetyl-D-glucosamine
-
Nostoc punctiforme N-acetyl-D-mannosamine
-
r
N-acetyl-D-glucosamine
-
Acaryochloris marina N-acetyl-D-mannosamine
-
r
N-acetyl-D-glucosamine
-
Nostoc sp. PCC 7120 N-acetyl-D-mannosamine
-
r
N-acetyl-D-glucosamine
-
Acaryochloris marina MBIC 11017 N-acetyl-D-mannosamine
-
r
N-acetyl-D-mannosamine
-
Trichormus variabilis N-acetyl-D-glucosamine
-
r
N-acetyl-D-mannosamine
-
Nostoc sp. N-acetyl-D-glucosamine
-
r
N-acetyl-D-mannosamine
-
Nostoc punctiforme N-acetyl-D-glucosamine
-
r
N-acetyl-D-mannosamine
-
Nostoc sp. PCC 7120 N-acetyl-D-glucosamine
-
r

Synonyms

Synonyms Comment Organism
AGE
-
Trichormus variabilis
AGE
-
Nostoc sp.
AGE
-
Nostoc punctiforme
AGE
-
Acaryochloris marina
N-acyl-D-glucosamine 2-epimerase
-
Trichormus variabilis
N-acyl-D-glucosamine 2-epimerase
-
Nostoc sp.
N-acyl-D-glucosamine 2-epimerase
-
Nostoc punctiforme
N-acyl-D-glucosamine 2-epimerase
-
Acaryochloris marina

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Trichormus variabilis
30
-
assay at Nostoc sp.
30
-
assay at Nostoc punctiforme

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Trichormus variabilis
7.5
-
assay at Nostoc sp.
7.5
-
assay at Nostoc punctiforme

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
14.7
-
N-acetylneuraminic acid pH 7.5, 30°C Nostoc sp.
15.1
-
N-acetylneuraminic acid pH 7.5, 30°C Nostoc punctiforme
38.7
-
N-acetylneuraminic acid pH 7.5, 30°C Trichormus variabilis
412
-
pyruvate pH 7.5, 30°C Trichormus variabilis
454
-
pyruvate pH 7.5, 30°C Nostoc sp.
463
-
pyruvate pH 7.5, 30°C Nostoc punctiforme

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.17
-
N-acetyl-D-glucosamine pH 7.5, 30°C Trichormus variabilis
3.15
-
N-acetyl-D-glucosamine pH 6.0, 30°C Trichormus variabilis
3.26
-
N-acetyl-D-glucosamine pH 8.0, 30°C Trichormus variabilis
4.96
-
N-acetyl-D-glucosamine pH 7.0, 30°C Trichormus variabilis