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Literature summary for 4.1.2.13 extracted from

  • Aziz, I.; Rashid, N.; Ashraf, R.; Bashir, Q.; Imanaka, T.; Akhtar, M.
    Pcal_0111, a highly thermostable bifunctional fructose-1,6-bisphosphate aldolase/phosphatase from Pyrobaculum calidifontis (2017), Extremophiles, 21, 513-521 .
    View publication on PubMed
Search for more literature for 4.1.2.13

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21-CodonPlus(DE3)-RIL cells Pyrobaculum calidifontis
expression in Escherichia coli Pyrobaculum calidifontis
gene Pcal_0111, sequence comparisons, recombinant expression of mostly soluble enzyme in Escherichia coli strain BL21-CodonPlus(DE3)-RIL Pyrobaculum calidifontis

Inhibitors

Inhibitors Comment Organism Structure
ADP
-
 Pyrobaculum calidifontis
ATP
-
 Pyrobaculum calidifontis
additional information the enzyme does not show any inhibition in the presence of 10 mM AMP Pyrobaculum calidifontis
Sodium borohydride incubation with sodium borohydride in the presence of substrate results more than 80% of decrease in aldolase activity; more than 80% of decrease in aldolase activity in the presence of substrate. Negligible decrease in aldolase activity when Pcal_0111 and sodium borohydride are incubated in the absence of the substrate; results in more than 80% of decrease in aldolase activity of Pcal_0111 in the presence of substrate, also a negligible decrease in aldolase activity is observed when Pcal_0111 and sodium borohydride are incubated in the absence of the substrate Pyrobaculum calidifontis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten kinetics, from Arrhenius plots, activation energies for phosphatase and aldolase reactions are calculated 46.5 and 75.62 kJ/mol, respectively Pyrobaculum calidifontis
0.821
-
 D-fructose 1,6-bisphosphate pH 8.0, 25°C Pyrobaculum calidifontis
0.822
-
 D-fructose 1,6-bisphosphate at pH 6.0 and 25°C Pyrobaculum calidifontis
0.822
-
 D-fructose 1,6-bisphosphate recombinant enzyme, pH 8.0, 55°C Pyrobaculum calidifontis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ activates Pyrobaculum calidifontis
Mn2+ activates Pyrobaculum calidifontis
additional information residues involved in metal binding include Asp11, His18, Asp52, Asp53, Gln95, Asp132, Asp233, and Glu357 Pyrobaculum calidifontis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
44513
-
 calculated from sequence Pyrobaculum calidifontis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-fructose 1,6-bisphosphate Pyrobaculum calidifontis
-
 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate Pyrobaculum calidifontis JCM 11548
-
 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate Pyrobaculum calidifontis VA1
-
 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate Pyrobaculum calidifontis JGM 11548
-
 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate + H2O Pyrobaculum calidifontis
-
 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate + H2O Pyrobaculum calidifontis JCM 11548
-
 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate + H2O Pyrobaculum calidifontis VA1
-
 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate + H2O Pyrobaculum calidifontis JGM 11548
-
 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Pyrobaculum calidifontis A3MSD2
-
-
Pyrobaculum calidifontis JCM 11548 A3MSD2
-
-
Pyrobaculum calidifontis JGM 11548 A3MSD2
-
-
Pyrobaculum calidifontis VA1 A3MSD2
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pyrobaculum calidifontis
Ni-NTA column chromatography Pyrobaculum calidifontis
recombinant enzyme from Escherichia coli strain BL21-CodonPlus(DE3)-RIL by heat treatment at 80°C for 25 min, nickel affinity chromatography, and dialysis Pyrobaculum calidifontis

Source Tissue

Source Tissue Comment Organism Textmining
additional information the facultative aerobe that grows optimally at 90-95°C Pyrobaculum calidifontis
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.3
-
 pH 8.0, 25°C Pyrobaculum calidifontis
1.32
-
 purified recombinant enzyme, substrate D-fructose 1,6-bisphosphate, pH 8.0, 55°C Pyrobaculum calidifontis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-fructose 1,6-bisphosphate
-
 Pyrobaculum calidifontis glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate 100% activity Pyrobaculum calidifontis glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate
-
 Pyrobaculum calidifontis JCM 11548 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate 100% activity Pyrobaculum calidifontis JCM 11548 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate
-
 Pyrobaculum calidifontis VA1 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate 100% activity Pyrobaculum calidifontis VA1 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate
-
 Pyrobaculum calidifontis JGM 11548 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate 100% activity Pyrobaculum calidifontis JGM 11548 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate + H2O
-
 Pyrobaculum calidifontis glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate + H2O preferred substrate Pyrobaculum calidifontis glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate + H2O
-
 Pyrobaculum calidifontis JCM 11548 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate + H2O preferred substrate Pyrobaculum calidifontis JCM 11548 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate + H2O
-
 Pyrobaculum calidifontis VA1 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate + H2O preferred substrate Pyrobaculum calidifontis VA1 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate + H2O
-
 Pyrobaculum calidifontis JGM 11548 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
D-fructose 1,6-bisphosphate + H2O preferred substrate Pyrobaculum calidifontis JGM 11548 glycerone phosphate + D-glyceraldehyde 3-phosphate
-
 ?
additional information purified recombinant Pcal_0111 catalyzes both phosphatase and aldolase reactions with specific activity values of 4 U and 1.3 U, respectively. Although the enzyme can utilize several substrates for dephosphorylation, no phosphatase activity is detected with AMP, UTP, NADP, glucose 6-phosphate, ribose 5-phosphate, riboflavin 5-monophosphate, and pyridoxal 5-phosphate. Varying amounts of phosphatase activity are found when ATP, GTP, ADP, CTP, glyceraldehyde 3-phosphate, fructose 6-phosphate, fructose 1,6-bisphosphate, and phosphoribosyl diphosphate are used as substrates. Substrate binding residues include Tyr229, Lys232, and Tyr358 Pyrobaculum calidifontis ?
-
 ?
additional information purified recombinant Pcal_0111 catalyzes both phosphatase and aldolase reactions with specific activity values of 4 U and 1.3 U, respectively. Although the enzyme can utilize several substrates for dephosphorylation, no phosphatase activity is detected with AMP, UTP, NADP, glucose 6-phosphate, ribose 5-phosphate, riboflavin 5-monophosphate, and pyridoxal 5-phosphate. Varying amounts of phosphatase activity are found when ATP, GTP, ADP, CTP, glyceraldehyde 3-phosphate, fructose 6-phosphate, fructose 1,6-bisphosphate, and phosphoribosyl diphosphate are used as substrates. Substrate binding residues include Tyr229, Lys232, and Tyr358 Pyrobaculum calidifontis JCM 11548 ?
-
 ?
additional information purified recombinant Pcal_0111 catalyzes both phosphatase and aldolase reactions with specific activity values of 4 U and 1.3 U, respectively. Although the enzyme can utilize several substrates for dephosphorylation, no phosphatase activity is detected with AMP, UTP, NADP, glucose 6-phosphate, ribose 5-phosphate, riboflavin 5-monophosphate, and pyridoxal 5-phosphate. Varying amounts of phosphatase activity are found when ATP, GTP, ADP, CTP, glyceraldehyde 3-phosphate, fructose 6-phosphate, fructose 1,6-bisphosphate, and phosphoribosyl diphosphate are used as substrates. Substrate binding residues include Tyr229, Lys232, and Tyr358 Pyrobaculum calidifontis VA1 ?
-
 ?
additional information purified recombinant Pcal_0111 catalyzes both phosphatase and aldolase reactions with specific activity values of 4 U and 1.3 U, respectively. Although the enzyme can utilize several substrates for dephosphorylation, no phosphatase activity is detected with AMP, UTP, NADP, glucose 6-phosphate, ribose 5-phosphate, riboflavin 5-monophosphate, and pyridoxal 5-phosphate. Varying amounts of phosphatase activity are found when ATP, GTP, ADP, CTP, glyceraldehyde 3-phosphate, fructose 6-phosphate, fructose 1,6-bisphosphate, and phosphoribosyl diphosphate are used as substrates. Substrate binding residues include Tyr229, Lys232, and Tyr358 Pyrobaculum calidifontis JGM 11548 ?
-
 ?

Subunits

Subunits Comment Organism
? x * 44000, SDS-PAGE Pyrobaculum calidifontis
? x * 44513, calculated from amino acid sequence Pyrobaculum calidifontis
? x * 44513, sequence calculation Pyrobaculum calidifontis

Synonyms

Synonyms Comment Organism
bifunctional fructose-1,6-bisphosphate aldolase/phosphatase
-
 Pyrobaculum calidifontis
fructose 1,6-bisphosphatase/aldolase
-
 Pyrobaculum calidifontis
fructose-1,6-bisphosphate aldolase/phosphatase
-
 Pyrobaculum calidifontis
More cf. EC 3.1.3.11 Pyrobaculum calidifontis
Pcal_0111
-
 Pyrobaculum calidifontis
Pcal_0111 bifunctional fructose-1,6-bisphosphate aldolase/phosphatase Pyrobaculum calidifontis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
80
-
 the enzyme denatures in the presence of ATP Pyrobaculum calidifontis
90
-
-
 Pyrobaculum calidifontis
90
-
 fructose 1,6-bisphosphate aldolase Pyrobaculum calidifontis
90
-
 difference in optimal temperature of aldolase and phosphatase activity of the enzyme could be due to difference in heat stabilities of the substrates Pyrobaculum calidifontis
100
-
 difference in optimal temperature of aldolase and phosphatase activity of the enzyme could be due to difference in heat stabilities of the substrates Pyrobaculum calidifontis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
80
-
 purified recombinant enzyme, 24 h, completely stable Pyrobaculum calidifontis
80
-
 no significant change in activity even after 24 h. The enzyme denatures in the presence of ATP Pyrobaculum calidifontis
90 100 purified recombinant enzyme, half-life is 120 min, no significant change in the circular dichroism spectra of the protein up to 90°C Pyrobaculum calidifontis
100
-
 half-life: 120 min Pyrobaculum calidifontis
100
-
 recombinant enzyme is highly stable with a half-life of 120 min at 100°C Pyrobaculum calidifontis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
 Pyrobaculum calidifontis

pI Value

Organism Comment pI Value Maximum pI Value
Pyrobaculum calidifontis sequence calculation
-
 6.16
Pyrobaculum calidifontis calculated from sequence
-
 6.16
Pyrobaculum calidifontis calculated from amino acid sequence
-
 6.2

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.75
-
 at pH 6.0 and 25°C Pyrobaculum calidifontis ATP

General Information

General Information Comment Organism
evolution the archaeal enzyme belongs to the class V of fructose-1,6-bisphosphatases. Gene expression of class V FBPase is regulated at the transcription level. The substrate binding residues, including Tyr229, Lys232, and Tyr358, and the residues involved in metal binding, including Asp11, His18, Asp52, Asp53, Gln95, Asp132, Asp233, and Glu357 are completely conserved in all the archaeal FBPases Pyrobaculum calidifontis
additional information the active site of enzyme Pcal_0111 contains a lysine residue which makes Schiff base with carbonyl group of the substrate Pyrobaculum calidifontis