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Literature summary for 3.1.3.11 extracted from

  • Topaz, G.R.; Epiter-Smith, V.; Robolo, C.; Emad, M.; Ford, V.; Daley, J.; Byron, J.; Stieglitz, K.A.
    Characterization of recombinant fructose 1,6-bisphosphatase (FBPase) gene mutations evidence of inhibition/activation of FBPase protein by gene mutation (2019), Biosci. Rep., 39, BSR20180960 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Sus scrofa

Protein Variants

Protein Variants Comment Organism
K112A mutations in the AMP-binding site demonstrates an eight to nine-fold decrease in AMP inhibition Sus scrofa
L56A mutant enzyme (interfacial mutant) exhibits an increase in Ki for fructose-2,6-bisphosphate by approximately 5fold Sus scrofa
L73A mutant enzyme (interfacial mutant) exhibits an increase in Ki for fructose-2,6-bisphosphate by approximately 5fold Sus scrofa
M177A mutation reveals an approximate two to three-fold decrease in inhibitory constants (Ki's) for natural inhibitors AMP and fructose-2,6-bisphosphate compared with the wild-type enzyme Sus scrofa
M248D the active site mutant enzyme displays an approximate 7fold increase in Ki for fructose-2,6-bisphosphate. The mutant enzyme displays a four-fold decrease in its apparent Michelis constant, and a six-fold increase in catalytic efficiency Sus scrofa
Y113A mutations in the AMP-binding site demonstrates an eight to nine-fold decrease in AMP inhibition Sus scrofa
Y164A mutation reveals an approximate two to three-fold decrease in inhibitory constants (Ki's) for natural inhibitors AMP and fructose-2,6-bisphosphate compared with the wild-type enzyme Sus scrofa

Inhibitors

Inhibitors Comment Organism Structure
AMP
-
Sus scrofa
fructose 2,6-bisphosphate
-
Sus scrofa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00125
-
D-fructose 1,6-bisphosphate mutant enzyme M248D, pH 7.5, 37°C Sus scrofa
0.0025
-
D-fructose 1,6-bisphosphate mutant enzyme Y113A, pH 7.5, 37°C Sus scrofa
0.00325
-
D-fructose 1,6-bisphosphate mutant enzyme K112A, pH 7.5, 37°C Sus scrofa
0.0042
-
D-fructose 1,6-bisphosphate mutant enzyme M177A, pH 7.5, 37°C Sus scrofa
0.0048
-
D-fructose 1,6-bisphosphate wild-type enzyme, pH 7.5, 37°C Sus scrofa
0.0051
-
D-fructose 1,6-bisphosphate mutant enzyme L73A, pH 7.5, 37°C Sus scrofa
0.0052
-
D-fructose 1,6-bisphosphate mutant enzyme L56A, pH 7.5, 37°C Sus scrofa
0.0053
-
D-fructose 1,6-bisphosphate mutant enzyme Y164A, pH 7.5, 37°C Sus scrofa

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-fructose 1,6-bisphosphate + H2O Sus scrofa
-
D-fructose 6-phosphate + phosphate
-
r

Organism

Organism UniProt Comment Textmining
Sus scrofa P00636
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Sus scrofa

Source Tissue

Source Tissue Comment Organism Textmining
kidney
-
Sus scrofa
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-fructose 1,6-bisphosphate + H2O
-
Sus scrofa D-fructose 6-phosphate + phosphate
-
r

Synonyms

Synonyms Comment Organism
FBPase
-
Sus scrofa
fructose-1,6-bisphosphatase
-
Sus scrofa

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
melting point of mutant enzyme Y113A, without ligand Sus scrofa
51
-
melting point of mutant enzyme K112A, without ligand Sus scrofa
54.2
-
melting point of mutant enzyme M177A, without ligand Sus scrofa
55.4
-
melting point of mutant enzyme Y164A, without ligand Sus scrofa
55.8
-
melting point of mutant enzyme L73A, without ligand Sus scrofa
56
-
melting point of mutant enzyme L56A, without ligand Sus scrofa
56
-
melting point of wild-type enzyme, without ligand Sus scrofa
62
-
melting point of mutant enzyme M248D, without ligand Sus scrofa

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
12.8
-
D-fructose 1,6-bisphosphate mutant enzyme M177A, pH 7.5, 37°C Sus scrofa
13.5
-
D-fructose 1,6-bisphosphate mutant enzyme Y164A, pH 7.5, 37°C Sus scrofa
20.5
-
D-fructose 1,6-bisphosphate wild-type enzyme, pH 7.5, 37°C Sus scrofa
22.5
-
D-fructose 1,6-bisphosphate mutant enzyme L73A, pH 7.5, 37°C Sus scrofa
25.6
-
D-fructose 1,6-bisphosphate mutant enzyme L56A, pH 7.5, 37°C Sus scrofa
26.8
-
D-fructose 1,6-bisphosphate mutant enzyme K112A, pH 7.5, 37°C Sus scrofa
27.2
-
D-fructose 1,6-bisphosphate mutant enzyme Y113A, pH 7.5, 37°C Sus scrofa
30.5
-
D-fructose 1,6-bisphosphate mutant enzyme M248D, pH 7.5, 37°C Sus scrofa

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.00015
-
fructose 2,6-bisphosphate mutant enzyme Y164A, pH 7.5, 37°C Sus scrofa
0.00017
-
fructose 2,6-bisphosphate mutant enzyme M177A, pH 7.5, 37°C Sus scrofa
0.00024
-
fructose 2,6-bisphosphate mutant enzyme K112A, pH 7.5, 37°C Sus scrofa
0.00025
-
fructose 2,6-bisphosphate mutant enzyme Y113A, pH 7.5, 37°C Sus scrofa
0.00025
-
fructose 2,6-bisphosphate wild-type enzyme, pH 7.5, 37°C Sus scrofa
0.00125
-
fructose 2,6-bisphosphate mutant enzyme L56A, pH 7.5, 37°C Sus scrofa
0.00135
-
fructose 2,6-bisphosphate mutant enzyme L73A, pH 7.5, 37°C Sus scrofa
0.0015
-
fructose 2,6-bisphosphate mutant enzyme M248D, pH 7.5, 37°C Sus scrofa

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.00075
-
mutant enzyme M177A, pH 7.5, 37°C Sus scrofa fructose 2,6-bisphosphate
0.00095
-
mutant enzyme Y164A, pH 7.5, 37°C Sus scrofa fructose 2,6-bisphosphate
0.0015
-
wild-type enzyme, pH 7.5, 37°C Sus scrofa fructose 2,6-bisphosphate
0.0015
-
mutant enzyme K112A, pH 7.5, 37°C Sus scrofa fructose 2,6-bisphosphate
0.0018
-
mutant enzyme Y113A, pH 7.5, 37°C Sus scrofa fructose 2,6-bisphosphate
0.0025
-
mutant enzyme Y164A, pH 7.5, 37°C Sus scrofa AMP
0.0035
-
mutant enzyme M177A, pH 7.5, 37°C Sus scrofa AMP
0.00695
-
mutant enzyme L73A, pH 7.5, 37°C Sus scrofa AMP
0.0072
-
mutant enzyme M248D, pH 7.5, 37°C Sus scrofa AMP
0.00725
-
mutant enzyme L73A, pH 7.5, 37°C Sus scrofa fructose 2,6-bisphosphate
0.00735
-
mutant enzyme L56A, pH 7.5, 37°C Sus scrofa AMP
0.0075
-
wild-type enzyme, pH 7.5, 37°C Sus scrofa AMP
0.0075
-
mutant enzyme L56A, pH 7.5, 37°C Sus scrofa fructose 2,6-bisphosphate
0.0105
-
mutant enzyme M248D, pH 7.5, 37°C Sus scrofa fructose 2,6-bisphosphate
0.0486
-
mutant enzyme Y113A, pH 7.5, 37°C Sus scrofa AMP
0.0685
-
mutant enzyme K112A, pH 7.5, 37°C Sus scrofa AMP