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Literature summary extracted from

  • Yep, A.; McLeish, M.J.
    Engineering the substrate binding site of benzoylformate decarboxylase (2009), Biochemistry, 48, 8387-8395.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.1.7 expressed in Escherichia coli BL21(DE3)pLysS cells Pseudomonas putida

Protein Variants

EC Number Protein Variants Comment Organism
4.1.1.7 A460Y the mutant shows significantly improved catalytic activity toward pyruvate Pseudomonas putida
4.1.1.7 T377L the mutant shows significantly improved catalytic activity toward pyruvate Pseudomonas putida
4.1.1.7 T377L/A460Y the double mutant shows significantly improved catalytic activity toward pyruvate (11000fold improvement in the ratio between pyruvate and benzoylformate utilization) Pseudomonas putida

Inhibitors

EC Number Inhibitors Comment Organism Structure
4.1.1.7 (R)-mandelate substrate analogue inhibitor Pseudomonas putida

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.1.1.7 0.04
-
 benzylformate mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 0.1
-
 2-ketopentanoate mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 0.15
-
 2-ketohexanoate mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 0.27
-
 benzylformate wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 0.28
-
 2-ketopentanoate mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 0.28
-
 2-ketohexanoate mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 0.42
-
 2-ketobutanoate mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 0.45
-
 benzylformate mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 1.1
-
 2-ketopentanoate mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 1.3
-
 2-ketobutanoate mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 1.3
-
 2-ketohexanoate mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 1.5
-
 benzylformate mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 2
-
 pyruvate mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 4.1
-
 2-ketohexanoate wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 4.3
-
 2-ketobutanoate mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 4.8
-
 pyruvate mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 6
-
 2-ketopentanoate wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 7.5
-
 2-ketobutanoate wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 21
-
 pyruvate mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida

Organism

EC Number Organism UniProt Comment Textmining
4.1.1.1 Zymomonas mobilis P06672
-
-
4.1.1.7 Pseudomonas putida P20906
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.1.7
-
 Pseudomonas putida

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.1.1 additional information PDC has no activity with benzoylformate Zymomonas mobilis ?
-
 ?
4.1.1.1 pyruvate
-
 Zymomonas mobilis acetaldehyde + CO2
-
 ?
4.1.1.7 2-ketohexanoate
-
 Pseudomonas putida valeraldehyde + CO2
-
 ?
4.1.1.7 2-ketopentanoate
-
 Pseudomonas putida butyraldehyde + CO2
-
 ?
4.1.1.7 2-oxobutanoate
-
 Pseudomonas putida propionaldehyde + CO2
-
 ?
4.1.1.7 benzylformate
-
 Pseudomonas putida benzaldehyde + CO2
-
 ?
4.1.1.7 pyruvate pyruvate is no substrate for the wild type enzyme Pseudomonas putida acetaldehyde + CO2
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.1.1.1 PDC
-
 Zymomonas mobilis
4.1.1.7 BFDC
-
 Pseudomonas putida

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.1.1.7 0.81
-
 2-ketopentanoate mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 1.1
-
 2-ketohexanoate mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 1.2
-
 2-ketohexanoate mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 1.4
-
 pyruvate mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 1.7
-
 benzylformate mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 2
-
 pyruvate mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 3.3
-
 2-ketopentanoate mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 3.6
-
 pyruvate mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 4
-
 2-ketobutanoate wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 4.4
-
 2-ketohexanoate mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 5.3
-
 2-ketohexanoate wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 5.8
-
 2-ketobutanoate mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 6
-
 2-ketobutanoate mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 6.2
-
 2-ketopentanoate mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 7.5
-
 2-ketobutanoate mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 11
-
 2-ketopentanoate wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 25
-
 benzylformate mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 110
-
 benzylformate mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 320
-
 benzylformate wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida

Cofactor

EC Number Cofactor Comment Organism Structure
4.1.1.1 thiamine diphosphate
-
 Zymomonas mobilis
4.1.1.7 thiamine diphosphate
-
 Pseudomonas putida

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.1.1.7 0.012
-
 pyruvate wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 0.065
-
 pyruvate mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 0.42
-
 pyruvate mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 0.5
-
 2-ketobutanoate wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 0.86
-
 2-ketohexanoate mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 1.3
-
 2-ketohexanoate wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 1.7
-
 2-ketobutanoate mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 1.8
-
 pyruvate mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 1.8
-
 2-ketopentanoate wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 3
-
 2-ketopentanoate mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 4.6
-
 2-ketobutanoate mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 8
-
 2-ketohexanoate mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 8.1
-
 2-ketopentanoate mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 14
-
 2-ketobutanoate mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 16
-
 2-ketohexanoate mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 16
-
 benzylformate mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 22
-
 2-ketopentanoate mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 43
-
 benzylformate mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 244
-
 benzylformate mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida
4.1.1.7 1180
-
 benzylformate wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C Pseudomonas putida