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Literature summary for 4.1.1.7 extracted from

  • Gocke, D.; Walter, L.; Gauchenova, E.; Kolter, G.; Knoll, M.; Berthold, C.L.; Schneider, G.; Pleiss, J.; Mueller, M.; Pohl, M.
    Rational protein design of ThDP-dependent enzymes-engineering stereoselectivity (2008), ChemBioChem, 9, 406-412.
    View publication on PubMed

Crystallization (Commentary)

Crystallization (Comment) Organism
mutant enzyme L461A, hanging drop vapour diffusion method Pseudomonas putida

Protein Variants

Protein Variants Comment Organism
A460G mutant exhibits a higher Km value compared to the wild type enzyme Pseudomonas putida
L461A the mutant catalyzes the S-stereoselective addition of larger acceptor aldehydes, such as propanal with benzaldehyde and its derivatives (a reaction not catalyzed by the wild type enzyme) Pseudomonas putida
L461G the mutant catalyzes the S-stereoselective addition of larger acceptor aldehydes, such as propanal with benzaldehyde and its derivatives (a reaction not catalyzed by the wild type enzyme) Pseudomonas putida
L461V mutant exhibits a higher Km value compared to the wild type enzyme Pseudomonas putida
P24A mutant exhibits a higher Km value compared to the wild type enzyme Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.06
-
benzoylformate mutant enzyme L461V, in potassium phosphate buffer (pH 6.5, 50 mM) Pseudomonas putida
0.25
-
benzoylformate mutant enzyme L461A, in potassium phosphate buffer (pH 6.5, 50 mM) Pseudomonas putida
0.37
-
benzoylformate wild type enzyme, in potassium phosphate buffer (pH 6.5, 50 mM) Pseudomonas putida
0.51
-
benzoylformate mutant enzyme P24A, in potassium phosphate buffer (pH 6.5, 50 mM) Pseudomonas putida
1.4
-
benzoylformate mutant enzyme L461G, in potassium phosphate buffer (pH 6.5, 50 mM) Pseudomonas putida
1.54
-
benzoylformate mutant enzyme A460G, in potassium phosphate buffer (pH 6.5, 50 mM) Pseudomonas putida

Organism

Organism UniProt Comment Textmining
Pseudomonas putida P20906
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography and Superdex G200 gel filtration Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
benzaldehyde + acetaldehyde
-
Pseudomonas putida (S)-2-hydroxy-1-phenylpropan-1-one
-
?
benzaldehyde + benzaldehyde
-
Pseudomonas putida (R)-benzoin
-
?
benzoylformate
-
Pseudomonas putida benzaldehyde + CO2
-
?
additional information the S-stereoselective addition of larger acceptor aldehydes, such as propanal with benzaldehyde and its derivatives is not catalyzed by the wild type enzyme, but with mutant enzyme L461A Pseudomonas putida ?
-
?

Synonyms

Synonyms Comment Organism
BFD
-
Pseudomonas putida

Cofactor

Cofactor Comment Organism Structure
thiamine diphosphate
-
Pseudomonas putida