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Literature summary for 4.1.1.7 extracted from

  • Andrews, F.; Wechsler, C.; Rogers, M.; Meyer, D.; Tittmann, K.; McLeish, M.
    Mechanistic and structural insight to an evolved benzoylformate decarboxylase with enhanced pyruvate decarboxylase activity (2016), Catalysts, 6, 190 .
No PubMed abstract available

Crystallization (Commentary)

Crystallization (Comment) Organism
mutant enzyme T377L/A460Y, hanging drop vapor diffusion method, using 0.1 M Tris (pH 8.5), 0.15 M CaCl2 and 22% PEG 400 (w/w) Pseudomonas putida

Protein Variants

Protein Variants Comment Organism
T377L/A460Y the mutant exhibits a greater than 10000fold increase in pyruvate/benzoylformate substrate utilization ratio compared to that of the wild type enzyme Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.14
-
4-methyl-2-oxopentanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
0.14
-
2-Oxohexanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
0.14
-
4-methylthio-2-oxobutanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
0.15
-
2-oxopentanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
0.27
-
benzoylformate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
0.39
-
2-oxobutanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
0.45
-
benzoylformate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
0.77
-
3-methyl-2-oxobutanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
0.82
-
phenylpyruvate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
1.2
-
3-methyl-2-oxopentanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
1.3
-
4-methylthio-2-oxobutanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
4.1
-
2-oxobutanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
5.3
-
2-Oxohexanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
9.6
-
4-methyl-2-oxopentanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
11
-
2-oxopentanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required, 1 mM used in assay conditions Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
benzoylformate Pseudomonas putida
-
benzaldehyde + CO2
-
?

Organism

Organism UniProt Comment Textmining
Pseudomonas putida P20906
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-oxobutanoate
-
Pseudomonas putida propionaldehyde + CO2
-
?
2-oxohexanoate
-
Pseudomonas putida valeraldehyde + CO2
-
?
2-oxopentanoate
-
Pseudomonas putida butyraldehyde + CO2
-
?
3-methyl-2-oxobutanoate
-
Pseudomonas putida 2-methylpropanal + CO2
-
?
3-methyl-2-oxopentanoate
-
Pseudomonas putida 2-methylbutanal + CO2
-
?
4-methyl-2-oxopentanoate
-
Pseudomonas putida 3-methylbutanal + CO2
-
?
4-methylthio-2-oxobutanoate
-
Pseudomonas putida 3-(methylsulfanyl)propanal + CO2
-
?
benzoylformate
-
Pseudomonas putida benzaldehyde + CO2
-
?
benzoylformate highest activity Pseudomonas putida benzaldehyde + CO2
-
?
additional information the enzyme exhibits very limited activity with pyruvate Pseudomonas putida ?
-
?
Phenylpyruvate
-
Pseudomonas putida Phenylacetaldehyde + CO2
-
?

Synonyms

Synonyms Comment Organism
BFDC
-
Pseudomonas putida

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1
-
2-oxopentanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
2
-
phenylpyruvate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
2.7
-
4-methyl-2-oxopentanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
4.1
-
2-Oxohexanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
4.5
-
4-methylthio-2-oxobutanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
4.6
-
4-methylthio-2-oxobutanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
5.9
-
2-Oxohexanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
6
-
2-oxobutanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
6
-
2-oxopentanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
7.5
-
2-oxobutanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
11
-
4-methyl-2-oxopentanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
12
-
3-methyl-2-oxobutanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
15
-
benzoylformate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
16
-
3-methyl-2-oxopentanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
320
-
benzoylformate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida

Cofactor

Cofactor Comment Organism Structure
thiamine diphosphate
-
Pseudomonas putida

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.1
-
phenylpyruvate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
0.5
-
2-oxobutanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
1.1
-
4-methyl-2-oxopentanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
1.3
-
2-Oxohexanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
1.6
-
3-methyl-2-oxopentanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
1.8
-
2-oxopentanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
2.5
-
phenylpyruvate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
2.6
-
3-methyl-2-oxobutanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
3.5
-
4-methylthio-2-oxobutanoate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida
7.5
-
2-oxopentanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
13
-
3-methyl-2-oxopentanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
15
-
3-methyl-2-oxobutanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
16
-
2-oxobutanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
20
-
4-methyl-2-oxopentanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
33
-
4-methylthio-2-oxobutanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
34
-
benzoylformate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
42
-
2-Oxohexanoate mutant enzyme T377L/A460Y, at pH 6.0 and 30°C Pseudomonas putida
1180
-
benzoylformate wild type enzyme, at pH 6.0 and 30°C Pseudomonas putida