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Literature summary for 2.6.1.16 extracted from

  • Czarnecka, J.; Kwiatkowska, K.; Gabriel, I.; Wojciechowski, M.; Milewski, S.
    Engineering Candida albicans glucosamine-6-phosphate synthase for efficient enzyme purification (2012), J. Mol. Recognit., 25, 564-570.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
overexpressed in Escherichia coli Candida albicans

Protein Variants

Protein Variants Comment Organism
additional information three recombinant versions containing internal oligoHis fragments are constructed: (a) by substituting residues 343-348 of the interdomain undecapeptide linker with hexaHis, (b) by replacing solvent-exposed residues 655-660 of the isomerase domain with hexaHis, and (c) by replacing amino acids at positions 568 and 569 with His residues to generate the three-dimensional hexaHis microdomain in the enzyme quaternary structure. The resulting constructs are purified to homogeneity. Catalytic properties are comparable with that of the wild-type enzyme. The construct containing the 655-660 hexaHis insert is found to be a homodimeric protein Candida albicans

Inhibitors

Inhibitors Comment Organism Structure
uridine 5'-diphospho-N-acetyl-D-glucosamine
-
Candida albicans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.12
-
D-fructose 6-phosphate mutant mutant Gfa1-His6DELTA655-660, pH 6.8, 37°C Candida albicans
1.33
-
L-glutamate mutant Gfa1-K568H/S569H, pH 6.8, 37°C Candida albicans
1.36
-
L-glutamate mutant Gfa1-His6DELTA343-348, pH 6.8, 37°C Candida albicans
1.41
-
L-glutamate wild-type, pH 6.8, 37°C Candida albicans
1.56
-
D-fructose 6-phosphate wild-type, pH 6.8, 37°C Candida albicans
1.9
-
D-fructose 6-phosphate mutant mutant Gfa1-K568H/S569H, pH 6.8, 37°C Candida albicans
2.36
-
L-glutamate mutant Gfa1-His6DELTA655-660, pH 6.8, 37°C Candida albicans
2.6
-
D-fructose 6-phosphate mutant Gfa1-His6DELTA343-348, pH 6.8, 37°C Candida albicans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
79000
-
SDS-PAGE, wild-type and mutants Candida albicans
160000
-
gel filtration, mutant Gfa1-His6DELTA655-660 Candida albicans
328000
-
gel filtration, mutant Gfa1-K568H/S569H Candida albicans
335000
-
gel filtration, mutant Gfa1-His6DELTA343-348 Candida albicans
340000
-
gel filtration, wildtype Candida albicans

Organism

Organism UniProt Comment Textmining
Candida albicans
-
-
-

Purification (Commentary)

Purification (Comment) Organism
using one-step immobilized metal-ion affinity chromatography Candida albicans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-glutamine + D-fructose 6-phosphate
-
Candida albicans L-glutamate + D-glucosamine 6-phosphate
-
?

Synonyms

Synonyms Comment Organism
glucosamine-6-phosphate synthase
-
Candida albicans

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
1.55
-
wild-type, pH 6.8, 37°C Candida albicans uridine 5'-diphospho-N-acetyl-D-glucosamine
1.7
-
mutant Gfa1-His6DELTA343-348, pH 6.8, 37°C Candida albicans uridine 5'-diphospho-N-acetyl-D-glucosamine
1.85
-
mutant Gfa1-His6DELTA655-660, pH 6.8, 37°C Candida albicans uridine 5'-diphospho-N-acetyl-D-glucosamine
2
-
mutant Gfa1-K568H/S569H, pH 6.8, 37°C Candida albicans uridine 5'-diphospho-N-acetyl-D-glucosamine