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Information on EC 2.6.1.16 - glutamine-fructose-6-phosphate transaminase (isomerizing) and Organism(s) Candida albicans and UniProt Accession P53704
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2.6.1.16 glutamine-fructose-6-phosphate transaminase (isomerizing)IUBMB Comments
Although the overall reaction is that of a transferase, the mechanism involves the formation of ketimine between fructose 6-phosphate and a 6-amino group from a lysine residue at the active site, which is subsequently displaced by ammonia (transamidination).
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Word Map
- 2.6.1.16
-
hexosamine
- udp-glcnac
- albicans
- n-acetylglucosamine
-
glcn6p
-
o-glcnac
- riboswitches
-
myasthenic
- udp-n-acetylglucosamine
-
o-linked
-
o-glcnacylation
- chitin
-
self-cleavage
- azaserine
-
self-cleaving
-
limb-girdle
- glutaminase
- o-glcnacase
- pyridostigmine
-
fatigable
-
glucosamine-induced
- d-glucosamine
-
hammerhead
- dpagt1
- 6-diazo-5-oxo-l-norleucine
- analysis
- synthesis
- medicine
The taxonomic range for the selected organisms is: Candida albicans
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
gfpt1, gfat1, glms ribozyme, glucosamine-6-phosphate synthase, glutamine:fructose-6-phosphate amidotransferase, glcn-6-p synthase, gfat2, glutamine fructose-6-phosphate amidotransferase, g-6-p synthase, glucosamine 6-phosphate synthase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
2-amino-2-deoxy-D-glucose-6-phosphate ketol-isomerase
-
-
-
-
GFAT
-
-
-
-
glucosamine 6-phosphate synthase
-
-
-
-
glucosamine 6-phosphate synthetase
-
-
-
-
glucosamine phosphate isomerase (glutamine-forming)
-
-
-
-
glucosamine synthase
-
-
-
-
glucosamine-6-phosphate isomerase (glutamine-forming)
-
-
-
-
glucosamine-6-phosphate synthetase
-
-
-
-
glucosamine:fructose-6-phosphate aminotransferase
-
-
-
-
glutamine-fructose 6-phosphate amidotransferase
-
-
-
-
glutamine-fructose 6-phosphate aminotransferase
-
-
-
-
glutamine:fructose-6-phosphate aminotransferase
-
-
-
-
hexosephosphate aminotransferase
-
-
-
-
isomerase, glucosamine phosphate (glutamine-forming)
-
-
-
-
L-glutamine fructose 6-phosphate transamidase
-
-
-
-
L-glutamine:D-fructose-6-phosphate amidotransferase (hexose isomerizing)
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
amino group transfer
-
-
-
-
PATHWAY SOURCE
PATHWAYS
BRENDA
-
-, -, -, -
SYSTEMATIC NAME
IUBMB Comments
L-glutamine:D-fructose-6-phosphate isomerase (deaminating)
Although the overall reaction is that of a transferase, the mechanism involves the formation of ketimine between fructose 6-phosphate and a 6-amino group from a lysine residue at the active site, which is subsequently displaced by ammonia (transamidination).
CAS REGISTRY NUMBER
COMMENTARY
9030-45-9
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
L-glutamine + D-fructose 6-phosphate
L-glutamate + D-glucosamine 6-phosphate
-
-
-
?
D-fructose 6-phosphate
D-glucose 6-phosphate
-
determination of hexose phosphate-isomerizing activity
-
-
?
L-gamma-glutamyl-p-nitroanilide + H2O
L-glutamine + p-nitroaniline
-
determination of amidohydrolysing activity
-
-
?
L-glutamine + D-fructose 6-phosphate
L-glutamate + D-glucosamine 6-phosphate
-
-
-
?
L-glutamine + D-fructose 6-phosphate
L-glutamate + D-glucosamine 6-phosphate
-
-
-
-
?
L-glutamine + D-fructose 6-phosphate
L-glutamate + D-glucosamine 6-phosphate
-
-
-
-
ir
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
L-glutamine + D-fructose 6-phosphate
L-glutamate + D-glucosamine 6-phosphate
-
-
-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
4-(1,3-dihydroxypropan-2-ylimino)-2-(4-hydroxyphenyl)chroman-5,7-diol
-
1-methyl 8-(2-oxopropyl) (2E,7S)-7-amino-4-oxooct-2-enedioate
-
ester derivative of N3-(4-metoxyfumaroyl)-(S)-2,3-diaminopropanoic acid, potent inhibitory activity against fungal glucosamine-6-phosphate synthase, good antifungal activity against Candida albicans
1-methyl 8-[(2R)-3-oxobutan-2-yl] (2E,7S)-7-amino-4-oxooct-2-enedioate
-
ester derivative of N3-(4-metoxyfumaroyl)-(S)-2,3-diaminopropanoic acid, potent inhibitory activity against fungal glucosamine-6-phosphate synthase, good antifungal activity against Candida albicans
8-(3,3-dimethyl-2-oxobutyl) 1-methyl (2E,7S)-7-amino-4-oxooct-2-enedioate
-
ester derivative of N3-(4-metoxyfumaroyl)-(S)-2,3-diaminopropanoic acid, potent inhibitory activity against fungal glucosamine-6-phosphate synthase, good antifungal activity against Candida albicans
methyl (2E)-4-([(2S)-2-amino-3-(methylamino)-3-oxopropyl]amino)-4-oxobut-2-enoate
-
-
N3-bromoacetyl-L-2,3-diaminopropanoic acid
-
competitive with respect to L-glutamine
N3-chloroacetyl-L-2,3-diaminopropanoic acid
-
competitive with respect to L-glutamine
N3-iodoacetyl-L-2,3-diaminopropanoic acid
-
competitive with respect to L-glutamine
additional information
synthesis of naringenin derivatives with potent glucosamine-6-phosphate synthase inhibitory capacities and antioxidant, antimicrobial, and preservative efficacy. Molecular docking and in silico ADMET analysis, structure-activity relationship studies, overview. MIC values for growth inhibition of the cells
-
additional information
-
synthesis of naringenin derivatives with potent glucosamine-6-phosphate synthase inhibitory capacities and antioxidant, antimicrobial, and preservative efficacy. Molecular docking and in silico ADMET analysis, structure-activity relationship studies, overview. MIC values for growth inhibition of the cells
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.67
L-gamma -glutamyl-p-nitroanilide
-
reaction mixture contains 1 mM L-gamma-glutamyl-p-nitroanilide, 1 mM EDTA, 1 mM DTT and the appropriately diluted enzyme preparation in 20 mM HEPES buffer (pH 7.5) at 25°C
0.34
L-glutamine
-
glutamine 6-phosphate-synthetic activity, wild-type, pH 7.5, 25°C
1.08
D-fructose 6-phosphate
-
hexose phosphate-isomerizing activity, wild-type with N-terminal His-tag, pH 7.5, 25°C
1.12
D-fructose 6-phosphate
-
mutant mutant Gfa1-His6DELTA655-660, pH 6.8, 37°C
1.2
D-fructose 6-phosphate
-
reaction mixture contains 2 mM D-fructose 6-phosphate, 0.5 mM NADP+, 1 mM EDTA, and appropriately diluted GlcN-6-P synthase preparation in 50 mM Tris/HCl buffer (pH 7.5) at 25°C
1.2
D-fructose 6-phosphate
-
wild type enzyme, in in 25 mM potassium phosphate buffer at pH 7.0 and 37°C
1.2
D-fructose 6-phosphate
-
hexose phosphate-isomerizing activity, wild-type, pH 7.5, 25°C
1.4
D-fructose 6-phosphate
-
hexose phosphate-isomerizing activity, residues 346-712, His-tagged hexose phosphate-isomerizing domain, pH 7.5, 25°C
1.41
D-fructose 6-phosphate
-
glutamine 6-phosphate-synthetic activity, wild-type, pH 7.5, 25°C
6.5
D-fructose 6-phosphate
-
hexose phosphate-isomerizing activity, wild-type with C-terminal His-tag, pH 7.5, 25°C
0.31
L-gamma-glutamyl-p-nitroanilide
-
residues 1-345, His-tagged glutamine amide-hydrolysing domain, pH 7.5, 25°C
0.64
L-gamma-glutamyl-p-nitroanilide
-
wild-type with C-terminal His-tag, pH 7.5, 25°C
12.2
L-gamma-glutamyl-p-nitroanilide
-
wild-type with N-terminal His-tag, pH 7.5, 25°C
0.34
L-glutamate
-
reaction mixture contains 10 mM L-glutamate, 1 mM EDTA, 1 mM DTT and an appropriately diluted enzyme preparation in 20 mM HEPES buffer (pH 7.5) at 25°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
12
L-glutamate
-
reaction mixture contains 10 mM L-glutamate, 1 mM EDTA, 1 mM DTT and an appropriately diluted enzyme preparation in 20 mM HEPES buffer (pH 7.5) at 25°C
12
L-glutamine
-
glutamine 6-phosphate-synthetic activity, wild-type, pH 7.5, 25°C
0.004
D-fructose 6-phosphate
-
hexose phosphate-isomerizing activity, wild-type with C-terminal His-tag, pH 7.5, 25°C
0.0092
D-fructose 6-phosphate
-
hexose phosphate-isomerizing activity, residues 346-712, His-tagged hexose phosphate-isomerizing domain, pH 7.5, 25°C
0.021
D-fructose 6-phosphate
-
hexose phosphate-isomerizing activity, wild-type with N-terminal His-tag, pH 7.5, 25°C
0.021
D-fructose 6-phosphate
-
hexose phosphate-isomerizing activity, wild-type, pH 7.5, 25°C
0.0213
D-fructose 6-phosphate
-
reaction mixture contains 2 mM D-fructose 6-phosphate, 0.5 mM NADP+, 1 mM EDTA, and appropriately diluted GlcN-6-P synthase preparation in 50 mM Tris/HCl buffer (pH 7.5) at 25°C
0.00092
L-gamma-glutamyl-p-nitroanilide
-
wild-type with N-terminal His-tag, pH 7.5, 25°C
0.0075
L-gamma-glutamyl-p-nitroanilide
-
residues 1-345, His-tagged glutamine amide-hydrolysing domain, pH 7.5, 25°C
0.0123
L-gamma-glutamyl-p-nitroanilide
-
reaction mixture contains 1 mM L-gamma-glutamyl-p-nitroanilide, 1 mM EDTA, 1 mM DTT and the appropriately diluted enzyme preparation in 20 mM HEPES buffer (pH 7.5) at 25°C
0.013
L-gamma-glutamyl-p-nitroanilide
-
wild-type with C-terminal His-tag, pH 7.5, 25°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0051
1-methyl 8-(2-oxopropyl) (2E,7S)-7-amino-4-oxooct-2-enedioate
Candida albicans
-
pH not specified in the publication, temperature not specified in the publication
0.0049
1-methyl 8-[(2R)-3-oxobutan-2-yl] (2E,7S)-7-amino-4-oxooct-2-enedioate
Candida albicans
-
pH not specified in the publication, temperature not specified in the publication
0.0052
8-(3,3-dimethyl-2-oxobutyl) 1-methyl (2E,7S)-7-amino-4-oxooct-2-enedioate
Candida albicans
-
pH not specified in the publication, temperature not specified in the publication
0.081
methyl (2E)-4-([(2S)-2,3-diamino-3-oxopropyl]amino)-4-oxobut-2-enoate
Candida albicans
-
pH 7.0, 37°C
0.127
methyl (2E)-4-([(2S)-2-amino-3-(methylamino)-3-oxopropyl]amino)-4-oxobut-2-enoate
Candida albicans
-
pH 7.0, 37°C
1.7
uridine 5'-diphospho-N-acetyl-D-glucosamine
Candida albicans
-
mutant Gfa1-His6DELTA343-348, pH 6.8, 37°C
1.85
uridine 5'-diphospho-N-acetyl-D-glucosamine
Candida albicans
-
mutant Gfa1-His6DELTA655-660, pH 6.8, 37°C
2
uridine 5'-diphospho-N-acetyl-D-glucosamine
Candida albicans
-
mutant Gfa1-K568H/S569H, pH 6.8, 37°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.021
-
glutamine 6-phosphate-synthetic activity, mutant V711F, pH 7.5, 25°C
0.095
-
amidohydrolysing activity of the mutant enzyme W97G using D-fructose 6-phosphate as substrate
0.256
-
amidohydrolysing activity of the mutant enzyme W97F using D-fructose 6-phosphate as substrate
0.89
-
amidohydrolysing activity of the wild type enzyme using D-fructose 6-phosphate as substrate
0.94
-
amidohydrolysing activity of the mutant enzyme V711F using D-fructose 6-phosphate as substrate
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
glucosamine-6-phosphate synthase is an important enzyme for bacterial and fungal cell wall synthesis
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
159000
-
hexose phosphate-isomerizing domain, Superdex 200 HR 10/30 gel filtration
39500
-
4 * 39500, hexose phosphate-isomerizing domain, Superdex 200 HR 10/30 gel filtration
42000
42000
-
isomerase domain consisting of residues 346-712, X-ray crystallography
42000
-
1 * 42000, glutamine amide-hydrolysing domain, Superdex 200 HR 10/30 gel filtration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homotetramer
monomer
-
1 * 42000, glutamine amide-hydrolysing domain, Superdex 200 HR 10/30 gel filtration
homotetramer
-
4 * 39500, hexose phosphate-isomerizing domain, Superdex 200 HR 10/30 gel filtration
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structure of the isomerase domain in complex with the allosteric inhibitor UDP-GlcNAc and in the presence of glucose 6-phosphate, fructose 6-phosphate and an analogue of the reaction intermediate, 2-amino-2-deoxy-D-mannitol 6-phosphate. Deduction of a solution structure of the native protein. The tetrameric enzyme can be described as a dimer of dimers, with each half similar to the related enzyme from Escherichia coli. The core of the protein consists of the isomerase domains
hanging drop vapour diffusion and sitting drop vapour diffusion in 20% (w/v) PEG 600 and 0.15 M KSCN at pH 8.5
-
molecular docking of inhibitors methyl (2E)-4-([(2S)-2,3-diamino-3-oxopropyl]amino)-4-oxobut-2-enoate and methyl (2E)-4-([(2S)-2-amino-3-(methylamino)-3-oxopropyl]amino)-4-oxobut-2-enoate
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
V711F
W97F
W97G
additional information
V711F
-
mutation results in an almost complete elimination of the GlcN-6-P-synthetic activity, with the retention of the amidohydrolase and sugar phosphate-isomerizing activities
V711F
-
reduction of the glucosamine 6-phosphate-synthetic activity, with the retention of the amidohydrolase and sugar phosphate-isomerizing activities
W97F
-
mutation results in an almost complete elimination of the GlcN-6-P synthetic activity, with the retention of the amidohydrolase and sugar phosphate-isomerizing activities
W97F
-
reduction of the glucosamine 6-phosphate-synthetic activity, with the retention of the amidohydrolase and sugar phosphate-isomerizing activities. Residue W97 functions as a molecular gate, opening and closing the intramolecular channel that connects the glutamine amide-hydrolysing and hexose phosphate-isomerizing domains
W97G
-
mutation results in an almost complete elimination of the GlcN-6-P-synthetic activity, with the retention of the amidohydrolase and sugar phosphate-isomerizing activities
W97G
-
almost complete elimination of the glucosamine 6-phosphate-synthetic activity, with the retention of the amidohydrolase and sugar phosphate-isomerizing activities. Residue W97 functions as a molecular gate, opening and closing the intramolecular channel that connects the glutamine amide-hydrolysing and hexose phosphate-isomerizing domains
additional information
-
expression of truncated enzyme variants as His-tagged proteins. Fragments encompassing residues 1-345 and 346-712 represent the functional glutamine amide-hydrolysing GAH and hexose phosphate-isomerizing domains ISOM, respectively. The native GAH domain is monomeric, whereas the native ISOM domain forms tetramers, as does the whole enzyme. The binding site for the feedback inhibitor, uridine 5'-diphospho-N-acetyl-D-glucosamine, is located in the ISOM domain. Inhibitor binding affects amidohydrolysing activity of the GAH domain and, as a consequence, the D-glucosamine-6-phosphate-synthetic activity of the whole enzyme. The fragment containing residues 218-283 is neither involved in ligand binding nor in protein oligomerization. An intramolecular channel connects the GAH and ISOM domains. The channel becomes leaky upon deletion of amino acids 709-712 and in the W97F and W97G mutants
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
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Ni2+-IDA agarose affinity chromatography and HiTrap desalting column chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Milewski, S.; Chmara, H.; Andruszkiewicz, R.; Borowski, E.
N3-haloacetyl derivatives of L-2,3-diaminopropanoic acid: novel inactivators of glucosamine-6-phosphate synthase
Biochim. Biophys. Acta
1115
225-229
1992
Bacillus pumilus, Candida albicans, Escherichia coli
Olchowy, J.; Jedrzejczak, R.; Milewski, S.; Rypniewski, W.
Crystallization and preliminary X-ray analysis of the isomerase domain of glucosamine-6-phosphate synthase from Candida albicans
Acta Crystallogr. Sect. F
61
994-996
2005
Candida albicans
Milewski, S.; Janiak, A.; Wojciechowski, M.
Structural analogues of reactive intermediates as inhibitors of glucosamine-6-phosphate synthase and phosphoglucose isomerase
Arch. Biochem. Biophys.
450
39-49
2006
Candida albicans
Olchowy, J.; Gabriel, I.; Milewski, S.
Functional domains and interdomain communication in Candida albicans glucosamine-6-phosphate synthase
Biochem. J.
404
121-130
2007
Candida albicans
Olchowy, J.; Kur, K.; Sachadyn, P.; Milewski, S.
Construction, purification, and functional characterization of His-tagged Candida albicans glucosamine-6-phosphate synthase expressed in Escherichia coli
Protein Expr. Purif.
46
309-315
2006
Candida albicans
Raczynska, J.; Olchowy, J.; Konariev, P.V.; Svergun, D.I.; Milewski, S.; Rypniewski, W.
The crystal and solution studies of glucosamine-6-phosphate synthase from Candida albicans
J. Mol. Biol.
372
672-688
2007
Candida albicans (P53704), Candida albicans
Czarnecka, J.; Kwiatkowska, K.; Gabriel, I.; Wojciechowski, M.; Milewski, S.
Engineering Candida albicans glucosamine-6-phosphate synthase for efficient enzyme purification
J. Mol. Recognit.
25
564-570
2012
Candida albicans
Pawlak, D.; Schielmann, M.; Wojciechowski, M.; Andruszkiewicz, R.
Synthesis and biological activity of novel ester derivatives of N(3)-(4-metoxyfumaroyl)-(S)-2,3-diaminopropanoic acid containing amide and keto function as inhibitors of glucosamine-6-phosphate synthase
Bioorg. Med. Chem. Lett.
26
3586-3589
2016
Candida albicans, Candida albicans ATCC 10231
Pawlak, D.; Stolarska, M.; Wojciechowski, M.; Andruszkiewicz, R.
Synthesis, anticandidal activity of N(3)-(4-methoxyfumaroyl)-(S)-2,3-diaminopropanoic amide derivatives - novel inhibitors of glucosamine-6-phosphate synthase
Eur. J. Med. Chem.
90
577-582
2015
Candida albicans
Lather, A.; Sharma, S.; Khatkar, A.
Naringenin derivatives as glucosamine-6-phosphate synthase inhibitors synthesis, antioxidants, antimicrobial, preservative efficacy, molecular docking and in silico ADMET analysis
BMC Chem.
14
41
2020
Aspergillus niger (A2QH83), Aspergillus niger, Proteus mirabilis (B4F0F0), Proteus mirabilis, Escherichia coli (P17169), Escherichia coli, Candida albicans (P53704), Candida albicans, Staphylococcus aureus (Q6GES3), Staphylococcus aureus, Pseudomonas aeruginosa (Q9HT25), Pseudomonas aeruginosa ATCC 15692 (Q9HT25), Staphylococcus aureus MRSA252 (Q6GES3), Proteus mirabilis HI4320 (B4F0F0), Pseudomonas aeruginosa 1C (Q9HT25), Candida albicans ATCC MYA-2876 (P53704), Pseudomonas aeruginosa PRS 101 (Q9HT25), Aspergillus niger FGSC A1513 (A2QH83), Pseudomonas aeruginosa DSM 22644 (Q9HT25), Pseudomonas aeruginosa CIP 104116 (Q9HT25), Pseudomonas aeruginosa LMG 12228 (Q9HT25), Aspergillus niger CBS 513.88 (A2QH83), Pseudomonas aeruginosa JCM 14847 (Q9HT25)
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