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Literature summary for 1.3.3.4 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)

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

Protein Variants

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

Inhibitors

Inhibitors Comment Organism Structure
acifluorfen
-
Bacillus subtilis
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]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Bacillus subtilis P32397
-
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
protoporphyrinogen IX + 3 O2
-
Bacillus subtilis protoporphyrin + 3 H2O2
-
?
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

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

Synonyms

Synonyms Comment Organism
PPO
-
Bacillus subtilis
protoporphyrinogen IX oxidase
-
Bacillus subtilis

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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

Cofactor

Cofactor Comment Organism Structure
FAD
-
Bacillus subtilis

Ki Value [mM]

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

IC50 Value

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

General Information

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