Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Blume, A.; Ghaderi, D.; Liebich, V.; Hinderlich, S.; Donner, P.; Reutter, W.; Lucka, L.
    UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase, functionally expressed in and purified from Escherichia coli, yeast, and insect cells (2004), Protein Expr. Purif., 35, 387-396.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.60 expressed with or without His-tag in Escherichia coli BL21 Star(DE3), Saccharomyces cerevisiae, Pichia pastoris, High five, Sf9, and Sf-900 cells, low yield in Escherichia coli and yeast, high expression of enzyme in High five, and Sf-900 cells Rattus norvegicus
3.2.1.183 functionally expressed in Escherichia coli, the yeast strains Saccharomyces cerevisiae and Pichia pastoris, and insect cells. In Escherichia coli, up to 2mg protein/l cell culture is expressed, in yeast cells 0.4mg/l, up to 100 mg/l, are expressed in insect cells. In all three cell systems, insoluble protein aggregates are also observed Rattus norvegicus
5.1.3.14 functional expression in Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris or insect cells. In all cell types, the expressed enzyme displayes both epimerase and kinase activities. In Escherichia coli up to 2 mg protein/l cell culture is expressed in yeast cells only 0.4 mg/ml, in insect cells up to 100 mg/L. In all three cell systems, insoluble protein aggregates are also observed. Expression and purification of the UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase in Sf-900 cells can yield the miligram amount of protein required for structural characterization of the enzyme. The easier expression in Escherichia coli and yeast provides sufficient quantities for enzymatic and kinetic characterization Rattus norvegicus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.1.60 79000
-
 6 * 79000 Rattus norvegicus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.60 ATP + N-acyl-D-mannosamine Rattus norvegicus key enzyme in the synthesis of N-acetylneuraminic acid and therefore of nearly all other sialic acids ADP + N-acyl-D-mannosamine 6-phosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.60 Rattus norvegicus
-
-
-
3.2.1.183 Rattus norvegicus
-
-
-
5.1.3.14 Rattus norvegicus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.60 recombinant enzymes from all species, purification from Sf-900 can yield milligram amounts of soluble, active protein Rattus norvegicus
3.2.1.183 purification from Escherichia coli results in 100 microgram/l pure and structurally intact protein. For insect cells, purification methods are established which result in up to 50 mg/l pure, soluble, and active protein Rattus norvegicus
5.1.3.14 enzyme expressed in Escherichia coli and enzyme expressed in insect cells Rattus norvegicus

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.7.1.60 liver
-
 Rattus norvegicus
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.1.60 10.9
-
 purified enzyme from Sf-900 cells, pH 8.1, 37°C Rattus norvegicus
5.1.3.14 10.9
-
 enzyme expressed in Sf-900 insect cells and purified Rattus norvegicus

Storage Stability

EC Number Storage Stability Organism
2.7.1.60 4°C, enzyme can be stored without substantial loss of activity Rattus norvegicus
5.1.3.14 4°C, purified enzyme expressed in insect cells, stable Rattus norvegicus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.60 ATP + N-acyl-D-mannosamine
-
 Rattus norvegicus ADP + N-acyl-D-mannosamine 6-phosphate
-
 ?
2.7.1.60 ATP + N-acyl-D-mannosamine key enzyme in the synthesis of N-acetylneuraminic acid and therefore of nearly all other sialic acids Rattus norvegicus ADP + N-acyl-D-mannosamine 6-phosphate
-
 ?
3.2.1.183 UDP-N-acetyl-alpha-D-glucosamine + H2O
-
 Rattus norvegicus ?
-
 ?
5.1.3.14 UDP-N-acetyl-D-glucosamine
-
 Rattus norvegicus UDP-N-acetyl-D-mannosamine
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.1.60 hexamer 6 * 79000 Rattus norvegicus

Synonyms

EC Number Synonyms Comment Organism
2.7.1.60 UDP-GlcNAc 2-epimerase/ManNAc kinase
-
 Rattus norvegicus
2.7.1.60 UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase two different enzyme activities combined in a single bifunctional enzyme Rattus norvegicus
3.2.1.183 UDP-GlcNAc 2-epimerase
-
 Rattus norvegicus
5.1.3.14 UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase
-
 Rattus norvegicus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.183 37
-
 assay at Rattus norvegicus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.183 7.5
-
 assay at Rattus norvegicus

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
2.7.1.60 1 2 pH above 12 results in almost complete disaggregation and complete loss of activity Rattus norvegicus
2.7.1.60 6
-
 pH below 6 results in almost complete disaggregation and complete loss of activity Rattus norvegicus