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Literature summary for 3.1.3.26 extracted from

  • Fu, D.; Li, Z.; Huang, H.; Yuan, T.; Shi, P.; Luo, H.; Meng, K.; Yang, P.; Yao, B.
    Catalytic efficiency of HAP phytases is determined by a key residue in close proximity to the active site (2011), Appl. Microbiol. Biotechnol., 90, 1295-1302.
    View publication on PubMed
Search for more literature for 3.1.3.26

Cloned(Commentary)

Cloned (Comment) Organism
gene APPA, DNA and amino acid sequence determination and analysis, expression of wild-type and mutant enzymes in Pichia pastoris strain GS115 Escherichia coli

Protein Variants

Protein Variants Comment Organism
G73D site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme Escherichia coli
G73E site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme Escherichia coli
G73L site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme Escherichia coli
G73R site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme Escherichia coli
G73S site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme Escherichia coli
G73T site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme Escherichia coli
G73Y site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.35
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate recombinant wild-type enzyme, pH 4.5, 37°C Escherichia coli
0.36
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate recombinant enzyme mutant G73S, pH 4.5, 37°C Escherichia coli
0.53
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate recombinant enzyme mutant G73D, pH 4.5, 37°C Escherichia coli
0.61
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate recombinant enzyme mutant G73E, pH 4.5, 37°C Escherichia coli
0.67
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate recombinant enzyme mutant G73T, pH 4.5, 37°C Escherichia coli

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 Escherichia coli
-
 ? + phosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli P07102 gene APPA
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Pichia pastoris strain GS115 by gel filtration to homogeneity Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
17
-
 purified recombinant enzyme mutant G73R, pH 4.5, 37°C Escherichia coli
74
-
 purified recombinant enzyme mutant G73Y, pH 4.5, 37°C Escherichia coli
220
-
 purified recombinant enzyme mutant G73L, pH 4.5, 37°C Escherichia coli
422
-
 purified recombinant enzyme mutant G73E, pH 4.5, 37°C Escherichia coli
513
-
 purified recombinant enzyme mutant G73T, pH 4.5, 37°C Escherichia coli
874
-
 purified recombinant enzyme mutant G73D, pH 4.5, 37°C Escherichia coli
2400
-
 purified recombinant enzyme mutant G73S, pH 4.5, 37°C Escherichia coli
3524
-
 purified recombinant wild-type enzyme, pH 4.5, 37°C Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
myo-inositol-1,2,3,4,5,6-hexakisphosphate + H2O
-
 Escherichia coli ? + phosphate
-
 ?

Subunits

Subunits Comment Organism
More molecular modeling using the crystal structure, overview Escherichia coli

Synonyms

Synonyms Comment Organism
AppA
-
 Escherichia coli
HAP phytase
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
 mutants G73R, G73T, G73Y, and G73D Escherichia coli
50
-
 wild-type enzyme and mutants G73E and G73L Escherichia coli
60
-
 mutant G73S Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5
-
 mutant G73D Escherichia coli
4
-
 mutants G73E and G73T Escherichia coli
4.5
-
 wild-type enzyme and mutants G73S, G73L, and G73Y Escherichia coli
5
-
 mutant G73R Escherichia coli

General Information

General Information Comment Organism
additional information catalytic efficiency of the HAP phytase is determined by key residue Arg79 located in close proximity to the active site Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
661
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate recombinant enzyme mutant G73T, pH 4.5, 37°C Escherichia coli
1199
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate recombinant enzyme mutant G73E, pH 4.5, 37°C Escherichia coli
1554
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate recombinant enzyme mutant G73D, pH 4.5, 37°C Escherichia coli
6017
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate recombinant enzyme mutant G73S, pH 4.5, 37°C Escherichia coli
6776
-
 myo-inositol-1,2,3,4,5,6-hexakisphosphate recombinant wild-type enzyme, pH 4.5, 37°C Escherichia coli