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

BRENDA support

Literature summary for 3.1.3.8 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
Search for more literature for 3.1.3.8

Cloned(Commentary)

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

Protein Variants

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

KM Value [mM]

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

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

Organism

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

Purification (Commentary)

Purification (Comment) Organism
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)

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

Synonyms

Synonyms Comment Organism
PhyA
-
 Aspergillus niger

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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

pH Range

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

kcat/KM [mM/s]

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