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Literature summary extracted from

  • Mullaney, E.J.; Locovare, H.; Sethumadhavan, K.; Boone, S.; Lei, X.G.; Ullah, A.H.
    Site-directed mutagenesis of disulfide bridges in Aspergillus niger NRRL 3135 phytase (PhyA), their expression in Pichia pastoris and catalytic characterization (2010), Appl. Microbiol. Biotechnol., 87, 1367-1372.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.3.8 expression in Pichia pastoris Aspergillus niger
3.1.3.8 gene phyA, expression in Pichia pastoris strain X33, subcloning in Escherichia coli strain JM109 and DH5alpha Aspergillus niger

Protein Variants

EC Number Protein Variants Comment Organism
3.1.3.8 C215S removal of disulfide bridge results in increase in Km, decrease in kcat value, loss of activity at 58°C Aspergillus niger
3.1.3.8 C215S site-directed mutagenesis, the mutant shows altered temperature optimum compared to the wild-type enzyme Aspergillus niger
3.1.3.8 C264G removal of disulfide bridge results in decrease in kcat value, loss of activity at 58°C Aspergillus niger
3.1.3.8 C264G site-directed mutagenesis, the mutant shows altered pH and temperature optima compared to the wild-type enzyme Aspergillus niger
3.1.3.8 C31G removal of disulfide bridge results in increase in Km, decrease in kcat value, loss of activity at 58°C Aspergillus niger
3.1.3.8 C31G site-directed mutagenesis, the mutant shows altered pH and temperature optima compared to the wild-type enzyme Aspergillus niger
3.1.3.8 C345G removal of disulfide bridge results in decrease in kcat value, loss of activity at 58°C Aspergillus niger
3.1.3.8 C435G site-directed mutagenesis, the mutant shows altered pH and temperature optima compared to the wild-type enzyme Aspergillus niger
3.1.3.8 C71S site-directed mutagenesis, inactive mutant Aspergillus niger
3.1.3.8 C71S removal of disulfide bridge results in loss of activity Aspergillus niger

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.3.8 0.045
-
 myo-inositol hexakisphosphate mutant C435G, pH 2.5, 42°C Aspergillus niger
3.1.3.8 0.045
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate pH 2.5, 42°C, recombinant mutant C435G Aspergillus niger
3.1.3.8 0.056
-
 myo-inositol hexakisphosphate mutant C264G, pH 6.0, 37°C Aspergillus niger
3.1.3.8 0.056
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate pH 6.0, 37°C, recombinant mutant C264G Aspergillus niger
3.1.3.8 0.07
-
 myo-inositol hexakisphosphate wild-type, pH 5.0, 58°C Aspergillus niger
3.1.3.8 0.07
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate pH 5.0, 58°C, recombinant wild-type enzyme Aspergillus niger
3.1.3.8 0.145
-
 myo-inositol hexakisphosphate mutant C215S, pH 5.0, 37°C Aspergillus niger
3.1.3.8 0.145
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate pH 5.0, 37°C, recombinant mutant C215S Aspergillus niger
3.1.3.8 0.18
-
 myo-inositol hexakisphosphate mutant C31G, pH 5.5, 53°C Aspergillus niger
3.1.3.8 0.18
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate pH 5.5, 53°C, recombinant mutant C31G Aspergillus niger

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.1.3.8 myo-inositol-1,2,3,4,5,6-hexakisphosphate + H2O Aspergillus niger
-
 ? + phosphate
-
 ?
3.1.3.8 myo-inositol-1,2,3,4,5,6-hexakisphosphate + H2O Aspergillus niger NRRL 3135
-
 ? + phosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.1.3.8 Aspergillus niger
-
 NRRL 3135
-
3.1.3.8 Aspergillus niger P34752 gene phyA
-
3.1.3.8 Aspergillus niger NRRL 3135
-
 NRRL 3135
-
3.1.3.8 Aspergillus niger NRRL 3135 P34752 gene phyA
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.3.8 recombinant wild-type enzyme mutant enzymes from Pichia pastoris strain X33 by ammonium sulfate fractionation, and two different steps of ion exchange chromatography Aspergillus niger

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.3.8 myo-inositol hexakisphosphate + H2O
-
 Aspergillus niger 1D-myo-inositol 1,2,4,5,6-pentakisphosphate + 1D-myo-inositol-1,2,5,6 tetrakisphosphate + phosphate
-
 ?
3.1.3.8 myo-inositol hexakisphosphate + H2O
-
 Aspergillus niger NRRL 3135 1D-myo-inositol 1,2,4,5,6-pentakisphosphate + 1D-myo-inositol-1,2,5,6 tetrakisphosphate + phosphate
-
 ?
3.1.3.8 myo-inositol-1,2,3,4,5,6-hexakisphosphate + H2O
-
 Aspergillus niger ? + phosphate
-
 ?
3.1.3.8 myo-inositol-1,2,3,4,5,6-hexakisphosphate + H2O
-
 Aspergillus niger NRRL 3135 ? + phosphate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.1.3.8 PhyA
-
 Aspergillus niger

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.1.3.8 37
-
 mutants C215S, C264G Aspergillus niger
3.1.3.8 37
-
 recombinant mutants C215S and C264G Aspergillus niger
3.1.3.8 42
-
 mutant C435G Aspergillus niger
3.1.3.8 42
-
 recombinant mutant C435G Aspergillus niger
3.1.3.8 53
-
 mutant C31G Aspergillus niger
3.1.3.8 53
-
 recombinant mutant C31G Aspergillus niger
3.1.3.8 58
-
 wild-type Aspergillus niger
3.1.3.8 58
-
 recombinant wild-type enzyme Aspergillus niger

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1.3.8 3 6 myo-inositol hexakisphosphate mutant C264G, pH 6.0, 37°C Aspergillus niger
3.1.3.8 3 6 myo-inositol-1,2,3,4,5,6-hexakisphosphate pH 6.0, 37°C, recombinant mutant C264G Aspergillus niger
3.1.3.8 62
-
 myo-inositol hexakisphosphate mutant C215S, pH 5.0, 37°C Aspergillus niger
3.1.3.8 62
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate pH 5.0, 37°C, recombinant mutant C215S Aspergillus niger
3.1.3.8 90
-
 myo-inositol hexakisphosphate mutant C435G, pH 2.5, 42°C Aspergillus niger
3.1.3.8 90
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate pH 2.5, 42°C, recombinant mutant C435G Aspergillus niger
3.1.3.8 234
-
 myo-inositol hexakisphosphate mutant C31G, pH 5.5, 53°C Aspergillus niger
3.1.3.8 234
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate pH 5.5, 53°C, recombinant mutant C31G Aspergillus niger
3.1.3.8 350
-
 myo-inositol hexakisphosphate wild-type, pH 5.0, 58°C Aspergillus niger
3.1.3.8 350
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate pH 5.0, 58°C, recombinant wild-type enzyme Aspergillus niger

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.3.8 2.5
-
 mutant C435G Aspergillus niger
3.1.3.8 2.5
-
 recombinant mutant C435G Aspergillus niger
3.1.3.8 5
-
 wild-type, mutant C215S Aspergillus niger
3.1.3.8 5
-
 recombinant wild-type enzyme and mutant C215S Aspergillus niger
3.1.3.8 5.5
-
 mutant C31G Aspergillus niger
3.1.3.8 5.5
-
 recombinant mutant C31G Aspergillus niger
3.1.3.8 6
-
 mutant C264G Aspergillus niger
3.1.3.8 6
-
 recombinant mutant C264G Aspergillus niger

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.1.3.8 additional information
-
 pH profiles of wild-type and mutant phytases PhyA, overview Aspergillus niger

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.1.3.8 0.00043
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate pH 5.0, 37°C, recombinant mutant C215S Aspergillus niger
3.1.3.8 0.00072
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate pH 6.0, 37°C, recombinant mutant C264G Aspergillus niger
3.1.3.8 0.0013
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate pH 5.5, 53°C, recombinant mutant C31G Aspergillus niger
3.1.3.8 0.002
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate pH 2.5, 42°C, recombinant mutant C435G Aspergillus niger
3.1.3.8 0.005
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate pH 5.0, 58°C, recombinant wild-type enzyme Aspergillus niger
3.1.3.8 430
-
 myo-inositol hexakisphosphate mutant C215S, pH 5.0, 37°C Aspergillus niger
3.1.3.8 720
-
 myo-inositol hexakisphosphate mutant C264G, pH 6.0, 37°C Aspergillus niger
3.1.3.8 1300
-
 myo-inositol hexakisphosphate mutant C31G, pH 5.5, 53°C Aspergillus niger
3.1.3.8 2000
-
 myo-inositol hexakisphosphate mutant C435G, pH 2.5, 42°C Aspergillus niger
3.1.3.8 5000
-
 myo-inositol hexakisphosphate wild-type, pH 5.0, 58°C Aspergillus niger