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

BRENDA support

Literature summary extracted from

  • Bansal, S.; Srivastava, A.; Mukherjee, G.; Pandey, R.; Verma, A.K.; Mishra, P.; Kundu, B.
    Hyperthermophilic asparaginase mutants with enhanced substrate affinity and antineoplastic activity: structural insights on their mechanism of action (2012), FASEB J., 26, 1161-1171.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.1 expression of His-tagged mutant enzymes in Escherichia coli strain Rosetta (DE3) Pyrococcus furiosus

Protein Variants

EC Number Protein Variants Comment Organism
3.5.1.1 K274E site-directed mutagenesis, the active site mutant shows improved enzymatic properties at physiological conditions compared to the wild-type. The mutant is thermodynamically stable and resistant to proteolytic digestion, displays no glutaminase activity, and shows increased and more significant killing of human cell lines HL60, MCF7, and K562 as compared to the Escherichia coli L-asparaginase Pyrococcus furiosus
3.5.1.1 T53Q site-directed mutagenesis, the active site mutant shows improved enzymatic properties at physiological conditions compared to the wild-type. The mutant is thermodynamically stable and resistant to proteolytic digestion, displays no glutaminase activity, and shows increased and more significant killing of human cell lines HL60, MCF7, and K562 as compared to the Escherichia coli L-asparaginase Pyrococcus furiosus
3.5.1.1 T53Q/K274E site-directed mutagenesis, the active site mutant shows improved enzymatic properties at physiological conditions compared to the wild-type. The mutant is thermodynamically stable and resistant to proteolytic digestion, displays no glutaminase activity, and shows increased and more significant killing of human cell lines HL60, MCF7, and K562 as compared to the Escherichia coli L-asparaginase Pyrococcus furiosus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.1 additional information
-
 additional information steady-state enzyme kinetics at different conditions of wild-type and mutant enzymes, overview Pyrococcus furiosus
3.5.1.1 2.1
-
 L-asparagine pH 8.2, 80°C, mutant K274E Pyrococcus furiosus
3.5.1.1 4.3
-
 L-asparagine pH 7.4, 37°C, mutant K274E Pyrococcus furiosus
3.5.1.1 5.41
-
 L-asparagine pH 7.4, 37°C, mutant T53Q/K274E Pyrococcus furiosus
3.5.1.1 7.52
-
 L-asparagine pH 7.4, 37°C, mutant T53Q Pyrococcus furiosus
3.5.1.1 8.12
-
 L-asparagine pH 7.4, 37°C, wild-type enzyme Pyrococcus furiosus
3.5.1.1 8.2
-
 L-asparagine pH 8.2, 80°C, mutant T53Q/K274E Pyrococcus furiosus
3.5.1.1 8.3
-
 L-asparagine pH 8.2, 80°C, mutant T53Q Pyrococcus furiosus
3.5.1.1 12.1
-
 L-asparagine pH 8.2, 80°C, wild-type enzyme Pyrococcus furiosus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.5.1.1 L-asparagine + H2O Pyrococcus furiosus
-
 L-aspartate + NH3
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.1 Pyrococcus furiosus Q8TZE8
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.1.1 recombinant His-tagged mutant enzymes from Escherichia coli strain Rosetta (DE3) by nickel affinity chromatography Pyrococcus furiosus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.1 L-asparagine + H2O
-
 Pyrococcus furiosus L-aspartate + NH3
-
 ?
3.5.1.1 additional information increased substrate accessibility through the active site loop plays a major role in determining activity, dynamic flipping of a critical Tyr residue is responsible for the activity of thermophilic L-asparaginases, molecular dynamic simulation and reaction mechanism, overview Pyrococcus furiosus ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.5.1.1 L-asparaginase
-
 Pyrococcus furiosus
3.5.1.1 PfA
-
 Pyrococcus furiosus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.1.1 37
-
 assay at in phosphate buffer Pyrococcus furiosus
3.5.1.1 80
-
 assay at in Tris-HCl buffer Pyrococcus furiosus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.1.1 17.98
-
 L-asparagine pH 7.4, 37°C, wild-type enzyme Pyrococcus furiosus
3.5.1.1 23.8
-
 L-asparagine pH 7.4, 37°C, mutant T53Q/K274E Pyrococcus furiosus
3.5.1.1 24.9
-
 L-asparagine pH 7.4, 37°C, mutant K274E Pyrococcus furiosus
3.5.1.1 35.2
-
 L-asparagine pH 7.4, 37°C, mutant T53Q Pyrococcus furiosus
3.5.1.1 199.2
-
 L-asparagine pH 8.2, 80°C, mutant K274E Pyrococcus furiosus
3.5.1.1 246.7
-
 L-asparagine pH 8.2, 80°C, mutant T53Q Pyrococcus furiosus
3.5.1.1 277.9
-
 L-asparagine pH 8.2, 80°C, mutant T53Q/K274E Pyrococcus furiosus
3.5.1.1 888.6
-
 L-asparagine pH 8.2, 80°C, wild-type enzyme Pyrococcus furiosus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.1.1 7.4
-
 assay at in phosphate buffer Pyrococcus furiosus
3.5.1.1 8.2
-
 assay at in Tris-HCl buffer Pyrococcus furiosus

General Information

EC Number General Information Comment Organism
3.5.1.1 additional information increased substrate accessibility through the active site loop plays a major role in determining activity, dynamic flipping of a critical Tyr residue is responsible for the activity of thermophilic L-asparaginases, molecular dynamic simulation and reaction mechanism, overview. Structure-function analysis of active site residues T53 and K274 Pyrococcus furiosus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.5.1.1 22.1
-
 L-asparagine pH 7.4, 37°C, wild-type enzyme Pyrococcus furiosus
3.5.1.1 30
-
 L-asparagine pH 8.2, 80°C, mutant T53Q Pyrococcus furiosus
3.5.1.1 34
-
 L-asparagine pH 8.2, 80°C, mutant T53Q/K274E Pyrococcus furiosus
3.5.1.1 44
-
 L-asparagine pH 7.4, 37°C, mutant T53Q/K274E Pyrococcus furiosus
3.5.1.1 47.2
-
 L-asparagine pH 7.4, 37°C, mutant T53Q Pyrococcus furiosus
3.5.1.1 57.8
-
 L-asparagine pH 7.4, 37°C, mutant K274E Pyrococcus furiosus
3.5.1.1 73
-
 L-asparagine pH 8.2, 80°C, wild-type enzyme Pyrococcus furiosus
3.5.1.1 96
-
 L-asparagine pH 8.2, 80°C, mutant K274E Pyrococcus furiosus