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Literature summary for 3.1.3.8 extracted from

  • Tran, T.; Mamo, G.; Búxo, L.; Le, N.; Gaber, Y.; Mattiasson, B.; Hatti-Kaul, R.
    Site-directed mutagenesis of an alkaline phytase: influencing specificity, activity and stability in acidic milieu (2011), Enzyme Microb. Technol., 49, 177-182.
    View publication on PubMed
Search for more literature for 3.1.3.8

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Bacillus sp. (in: Bacteria)

Protein Variants

Protein Variants Comment Organism
E180N site-directed mutagenesis, cell surface residue, the mutant shows activity similar to the wild-type enzyme Bacillus sp. (in: Bacteria)
E227S site-directed mutagenesis, active site residue, the mutant shows highly reduced activity compared to the wild-type enzyme Bacillus sp. (in: Bacteria)
E229V site-directed mutagenesis, cell surface residue, the mutant shows 19% increased activity compared to the wild-type enzyme Bacillus sp. (in: Bacteria)
K179R site-directed mutagenesis, active site residue, the mutant shows highly reduced activity compared to the wild-type enzyme Bacillus sp. (in: Bacteria)
K77R site-directed mutagenesis, active site residue, the mutant shows highly reduced activity compared to the wild-type enzyme Bacillus sp. (in: Bacteria)
K77R/K179R site-directed mutagenesis, mutation of residues not directly involved in the catalysis but involved in substrate binding, the double mutant phytase shows higher stability at pH 2.6-3.0 compared to the wild-type enzyme. The mutant retains over 80% of its initial activity after 3 h incubation at pH 2.6 while the wild-type phytase retains only about 40% of its original activity. The mutant shows highly reduced activity compared to the wild-type enzyme Bacillus sp. (in: Bacteria)
P257R site-directed mutagenesis, cell surface residue, the mutant shows reduced activity compared to the wild-type enzyme Bacillus sp. (in: Bacteria)
S283R site-directed mutagenesis, cell surface residue, the mutant shows 13% increased activity compared to the wild-type enzyme Bacillus sp. (in: Bacteria)

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ activates Bacillus sp. (in: Bacteria)

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
phytate + H2O Bacillus sp. (in: Bacteria)
-
 ?
-
 ?
phytate + H2O Bacillus sp. (in: Bacteria) MD2
-
 ?
-
 ?

Organism

Organism UniProt Comment Textmining
Bacillus sp. (in: Bacteria)
-
-
-
Bacillus sp. (in: Bacteria) MD2
-
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.3
-
 recombinant mutant E227S, pH 7.0, 70°C Bacillus sp. (in: Bacteria)
5.7
-
 recombinant mutant K179R, pH 7.0, 70°C Bacillus sp. (in: Bacteria)
7.3
-
 recombinant mutant K77R, pH 7.0, 70°C Bacillus sp. (in: Bacteria)
7.5
-
 recombinant mutant K177R/K179R, pH 7.0, 70°C Bacillus sp. (in: Bacteria)
18.5
-
 recombinant mutant P257R, pH 7.0, 70°C Bacillus sp. (in: Bacteria)
31.2
-
 recombinant mutant E180N, pH 7.0, 70°C Bacillus sp. (in: Bacteria)
32.2
-
 recombinant wild-type enzyme, pH 7.0, 70°C Bacillus sp. (in: Bacteria)
35.7
-
 recombinant mutant S283R, pH 7.0, 70°C Bacillus sp. (in: Bacteria)
37.8
-
 recombinant mutant E229V, pH 7.0, 70°C Bacillus sp. (in: Bacteria)

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl phosphate + H2O
-
 Bacillus sp. (in: Bacteria) 4-nitrophenol + phosphate
-
 ?
4-nitrophenyl phosphate + H2O
-
 Bacillus sp. (in: Bacteria) MD2 4-nitrophenol + phosphate
-
 ?
diphosphate + H2O Na-diphosphate Bacillus sp. (in: Bacteria) 2 phosphate
-
 ?
diphosphate + H2O Na-diphosphate Bacillus sp. (in: Bacteria) MD2 2 phosphate
-
 ?
additional information substrate specificities of wild-type and mutant enzymes, overview Bacillus sp. (in: Bacteria) ?
-
 ?
additional information substrate specificities of wild-type and mutant enzymes, overview Bacillus sp. (in: Bacteria) MD2 ?
-
 ?
phytate + H2O
-
 Bacillus sp. (in: Bacteria) ?
-
 ?
phytate + H2O Na- and Ca-phytate, the first results in higher activity, substrate binding analysis by molecular modelling of phytate inside the active site, residue K77 is able to form two hydrogen bonds with the two phosphate groups on the phytate, whereas in case of K77R, the guanidinium group of arginine is able to form up to 4 H-bonds with the two phosphate groups Bacillus sp. (in: Bacteria) ?
-
 ?
phytate + H2O
-
 Bacillus sp. (in: Bacteria) MD2 ?
-
 ?
phytate + H2O Na- and Ca-phytate, the first results in higher activity, substrate binding analysis by molecular modelling of phytate inside the active site, residue K77 is able to form two hydrogen bonds with the two phosphate groups on the phytate, whereas in case of K77R, the guanidinium group of arginine is able to form up to 4 H-bonds with the two phosphate groups Bacillus sp. (in: Bacteria) MD2 ?
-
 ?

Synonyms

Synonyms Comment Organism
alkaline phytase
-
 Bacillus sp. (in: Bacteria)

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
 assay at Bacillus sp. (in: Bacteria)

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5
-
 enzyme mutant E227S Bacillus sp. (in: Bacteria)
6
-
 wild-type enzyme Bacillus sp. (in: Bacteria)