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Literature summary extracted from

  • Sun, L.; Wen, X.; Tan, Y.; Li, H.; Yang, X.; Zhao, Y.; Wang, B.; Cao, Q.; Niu, C.; Xi, Z.
    Site-directed mutagenesis and computational study of the Y366 active site in Bacillus subtilis protoporphyrinogen oxidase (2009), Amino Acids, 37, 523-530.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.3.3.4 expression of wild-type and mutant enzymes in Escherichia coli strain XL-1 Blue in inclusion bodies Bacillus subtilis

Protein Variants

EC Number Protein Variants Comment Organism
1.3.3.4 Y366A site-directed mutagenesis, the mutant enzyme shows 10% of wild-type enzyme activity Bacillus subtilis
1.3.3.4 Y366E site-directed mutagenesis, the mutant enzyme shows 1% of wild-type enzyme activity Bacillus subtilis
1.3.3.4 Y366H site-directed mutagenesis, the mutant enzyme shows 10% of wild-type enzyme activity Bacillus subtilis

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.3.3.4 acifluorfen
-
 Bacillus subtilis
1.3.3.4 additional information Bacillus subtilis PPO is unique because of its resistance to inhibition by diphenylethers involving residue Y366 at a site adjacent to substrate and FAD cofactor, modeling and docking studies, overview Bacillus subtilis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.3.3.4 0.00067
-
 protoporphyrinogen IX pH 7.5, 22°C, recombinant wild-type enzyme Bacillus subtilis
1.3.3.4 0.00107
-
 protoporphyrinogen IX pH 7.5, 22°C, recombinant mutant Y366A Bacillus subtilis
1.3.3.4 0.00234
-
 protoporphyrinogen IX pH 7.5, 22°C, recombinant mutant Y366E Bacillus subtilis
1.3.3.4 0.00354
-
 protoporphyrinogen IX pH 7.5, 22°C, recombinant mutant Y366H Bacillus subtilis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.3.3.4 protoporphyrinogen IX + 3 O2 Bacillus subtilis
-
 protoporphyrin + 3 H2O2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.3.3.4 Bacillus subtilis P32397
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.3.3.4 recombinant wild-type and mutant enzymes from Escherichia coli strain XL-1 Blue inclusion bodies Bacillus subtilis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.3.3.4 additional information
-
-
 Bacillus subtilis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.3.4 protoporphyrinogen IX + 3 O2
-
 Bacillus subtilis protoporphyrin + 3 H2O2
-
 ?
1.3.3.4 protoporphyrinogen IX + 3 O2 Bacillus subtilis PPO shows a broad substrate specificity involving residue Y366 at a site adjacent to substrate and FAD cofactor, modeling and docking studies and structure-function relationship, overview Bacillus subtilis protoporphyrin + 3 H2O2
-
 ?

Subunits

EC Number Subunits Comment Organism
1.3.3.4 More modeling and docking studies and structure-function relationship, overview. Homology modelling and structure comparisons, overview Bacillus subtilis

Synonyms

EC Number Synonyms Comment Organism
1.3.3.4 PPO
-
 Bacillus subtilis
1.3.3.4 protoporphyrinogen IX oxidase
-
 Bacillus subtilis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.3.3.4 22
-
 assay at room temperature Bacillus subtilis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.3.3.4 0.00015
-
 protoporphyrinogen IX pH 7.5, 22°C, recombinant mutant Y366E Bacillus subtilis
1.3.3.4 0.00042
-
 protoporphyrinogen IX pH 7.5, 22°C, recombinant mutant Y366A Bacillus subtilis
1.3.3.4 0.0016
-
 protoporphyrinogen IX pH 7.5, 22°C, recombinant mutant Y366H Bacillus subtilis
1.3.3.4 0.0034
-
 protoporphyrinogen IX pH 7.5, 22°C, recombinant wild-type enzyme Bacillus subtilis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.3.3.4 7.5
-
 assay at Bacillus subtilis

Cofactor

EC Number Cofactor Comment Organism Structure
1.3.3.4 FAD
-
 Bacillus subtilis

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.3.3.4 0.029
-
 acifluorfen pH 7.5, 22°C, recombinant mutant Y366A Bacillus subtilis
1.3.3.4 0.033
-
 acifluorfen pH 7.5, 22°C, recombinant wild-type enzyme Bacillus subtilis
1.3.3.4 0.053
-
 acifluorfen pH 7.5, 22°C, recombinant mutant Y366H Bacillus subtilis
1.3.3.4 0.154
-
 acifluorfen pH 7.5, 22°C, recombinant mutant Y366E Bacillus subtilis

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
1.3.3.4 0.133
-
 pH 7.5, 22°C, recombinant mutant Y366H Bacillus subtilis acifluorfen
1.3.3.4 0.173
-
 pH 7.5, 22°C, recombinant mutant Y366A Bacillus subtilis acifluorfen
1.3.3.4 0.183
-
 pH 7.5, 22°C, recombinant wild-type enzyme Bacillus subtilis acifluorfen
1.3.3.4 0.617
-
 pH 7.5, 22°C, recombinant mutant Y366E Bacillus subtilis acifluorfen

General Information

EC Number General Information Comment Organism
1.3.3.4 metabolism protoporphyrinogen IX oxidase is the last common enzyme of heme and chlorophyll biosynthesis Bacillus subtilis