Information on EC 4.1.1.7 - benzoylformate decarboxylase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
4.1.1.7
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RECOMMENDED NAME
GeneOntology No.
benzoylformate decarboxylase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
phenylglyoxylate = benzaldehyde + CO2
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C-C bond formation
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carboligation
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condensation
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decarboxylation
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
4-hydroxymandelate degradation
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Aminobenzoate degradation
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mandelate degradation I
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Microbial metabolism in diverse environments
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4-hydroxymandelate degradation
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SYSTEMATIC NAME
IUBMB Comments
benzoylformate carboxy-lyase (benzaldehyde-forming)
A thiamine-diphosphate protein.
CAS REGISTRY NUMBER
COMMENTARY hide
9025-00-7
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain EBF 65/65
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
gene mdlC
SwissProt
Manually annotated by BRENDA team
strain LW-4
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Manually annotated by BRENDA team
strain PRS 1
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Manually annotated by BRENDA team
strain ST-201
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Manually annotated by BRENDA team
strain ST-201
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
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penultimate enzyme in the mandelate pathway, that catalyzes the non-oxidative decarboxylation of benzoylformate to yield carbon dioxide and benzaldehyde
physiological function
additional information
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the first step in catalysis by all ThDP-dependent enzymes is the deprotonation of the C2 carbon by the 4'-imino group to form the ylide. Both active sites contain two histidine residues in the catalytic dimer, three ionizable residues in the active site of the enzyme are Ser26, His70 and His281. His281 is the donor for the protonation of the enamine intermediate, Ser26 forms a hydrogen bond with the carboxylate of the substrate
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(E)-2-oxo-4(pyridin-3-yl)-3-butenoic acid
3-(pyridin-3-yl)acrylaldehyde + CO2
show the reaction diagram
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-
?
(p-methoxybenzoyl)formate
p-methoxybenzaldehyde + CO2
show the reaction diagram
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-
-
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(p-methylbenzoyl)formate
p-methylbenzaldehyde + CO2
show the reaction diagram
2 benzaldehyde
(R)-benzoin
show the reaction diagram
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i.e. (R)-2-hydroxy-1,2-diphenylethan-1-one
-
?
2-Fluoro-benzaldehyde + acetaldehyde
(S)-1-(2-Fluoro-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
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-
-
-
?
2-Fluoro-benzaldehyde + acetaldehyde
1-(2-Fluoro-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
-
91% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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-
?
2-fluorobenzaldehyde
(R)-1,2-Bis-(2-fluoro-phenyl)-2-hydroxy-ethanone
show the reaction diagram
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carboligation
68% yield and more than 99% enantiomeric excess of the (R)-enantiomer
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?
2-furaldehyde
(R)-1,2-di-furan-2-yl-2-hydroxy-ethanone
show the reaction diagram
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carboligation
62% yield and 94% enantiomeric excess of the (R)-enantiomer
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?
2-furaldehyde + acetaldehyde
(S)-1-Furan-2-yl-2-hydroxy-propan-1-one
show the reaction diagram
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?
2-ketohexanoate
valeraldehyde + CO2
show the reaction diagram
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?
2-ketopentanoate
butyraldehyde + CO2
show the reaction diagram
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?
2-Methyl-benzaldehyde + acetaldehyde
(S)-2-Hydroxy-1-(2-methylphenyl)-propanone
show the reaction diagram
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-
-
?
2-Methyl-benzaldehyde + acetaldehyde
2-Hydroxy-1-o-tolyl-propan-1-one
show the reaction diagram
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4% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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?
2-oxo-3-phenylpropanoic acid
phenylacetaldehyde + CO2
show the reaction diagram
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?
2-oxo-4-methylhexanoic acid
3-methylpentanal + CO2
show the reaction diagram
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?
2-oxo-4-methylpentanoic acid
3-methylbutanal + CO2
show the reaction diagram
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?
2-oxo-4-phenylbutanoic acid
3-phenylpropionaldehyde + CO2
show the reaction diagram
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?
2-oxo-5-phenylpentanoic acid
4-phenylpentanal + CO2
show the reaction diagram
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?
2-oxobutanoate
propionaldehyde + CO2
show the reaction diagram
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?
2-oxobutanoic acid
propionaldehyde + CO2
show the reaction diagram
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?
2-oxohexanoic acid
valeraldehyde + CO2
show the reaction diagram
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?
2-oxopentanoic acid
butyraldehyde + CO2
show the reaction diagram
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?
3,5-Difluoro-benzaldehyde + acetaldehyde
(S)-1-(3,5-Difluoro-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
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?
3,5-dimethoxybenzaldehyde + acetaldehyde
(S)-1-(3,5-Dimethoxy-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
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?
3-Benzyloxy-benzaldehyde + acetaldehyde
1-(3-Benzyloxy-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
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more than 99% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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?
3-Bromo-benzaldehyde + acetaldehyde
(S)-1-(3-Bromo-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
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?
3-Bromo-benzaldehyde + acetaldehyde
1-(3-Bromo-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
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96% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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?
3-Chloro-benzaldehyde + acetaldehyde
(S)-1-(3-Chloro-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
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?
3-Chloro-benzaldehyde + acetaldehyde
1-(3-Chloro-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
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94% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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?
3-ethoxy-4-hydroxybenzoylformate
ethyl vanillin + CO2
show the reaction diagram
3-Ethoxy-benzaldehyde + acetaldehyde
(S)-1-(3-Ethoxy-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
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?
3-Ethoxy-benzaldehyde + acetaldehyde
1-(3-Ethoxy-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
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97% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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?
3-Fluoro-benzaldehyde + acetaldehyde
(S)-1-(3-Fluoro-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
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-
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?
3-Fluoro-benzaldehyde + acetaldehyde
1-(3-Fluoro-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
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87% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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?
3-Formyl-benzonitrile + acetaldehyde
3-((S)-2-Hydroxy-propionyl)-benzonitrile
show the reaction diagram
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?
3-Formyl-benzonitrile + acetaldehyde
3-(2-Hydroxy-propionyl)-benzonitrile
show the reaction diagram
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92% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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?
3-Hydroxy-benzaldehyde + acetaldehyde
(S)-2-Hydroxy-1-(3-hydroxy-phenyl)-propan-1-one
show the reaction diagram
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?
3-Hydroxy-benzaldehyde + acetaldehyde
2-Hydroxy-1-(3-hydroxy-phenyl)-propan-1-one
show the reaction diagram
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92% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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?
3-Isopropoxy-benzaldehyde + acetaldehyde
(S)-2-Hydroxy-1-(3-isopropoxy-phenyl)-propan-1-one
show the reaction diagram
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?
3-Isopropoxy-benzaldehyde + acetaldehyde
2-Hydroxy-1-(3-isopropoxy-phenyl)-propan-1-one
show the reaction diagram
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more thahn 99% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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?
3-Methoxy-benzaldehyde + acetaldehyde
(S)-2-Hydroxy-1-(3-methoxy-phenyl)-propan-1-one
show the reaction diagram
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-
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?
3-Methoxy-benzaldehyde + acetaldehyde
2-Hydroxy-1-(3-methoxy-phenyl)-propan-1-one
show the reaction diagram
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96% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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?
3-methoxybenzaldehyde
(R)-2-Hydroxy-1,2-bis-(3-methoxy-phenyl)-ethanone
show the reaction diagram
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carboligation
18% yield and more than 99% enantiomeric excess of the (R)-enantiomer
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?
3-Methoxymethoxy-benzaldehyde + acetaldehyde
(S)-2-Hydroxy-1-(3-methoxymethoxy-phenyl)-propan-1-one
show the reaction diagram
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?
3-methyl-2-oxobutanoic acid
2-methylpropanal + CO2
show the reaction diagram
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?
3-methyl-2-oxopentanoic acid
2-methylbutanal + CO2
show the reaction diagram
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?
3-Methyl-benzaldehyde + acetaldehyde
(S)-2-Hydroxy-1-m-tolyl-propan-1-one
show the reaction diagram
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-
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?
3-Methyl-benzaldehyde + acetaldehyde
2-Hydroxy-1-m-tolyl-propan-1-one
show the reaction diagram
-
97% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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?
3-Phenoxy-benzaldehyde + acetaldehyde
2-Hydroxy-1-(3-phenoxy-phenyl)-propan-1-one
show the reaction diagram
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more than 99% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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?
4,4-dimethyl-2-oxopentanoic acid
3,3-dimethylbutanal + CO2
show the reaction diagram
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?
4-Bromo-benzaldehyde + acetaldehyde
1-(4-Bromo-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
-
83% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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?
4-bromobenzaldehyde
(R)-1,2-bis-(4-bromo-phenyl)-2-hydroxy-ethanone
show the reaction diagram
-
carboligation
13% yield and more than 99% enantiomeric excess of the (R)-enantiomer
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?
4-bromobenzaldehyde + acetaldehyde
(S)-1-(4-Bromo-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
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-
-
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?
4-Chloro-benzaldehyde + acetaldehyde
1-(4-Chloro-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
-
82% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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?
4-chlorobenzaldehyde
(R)-1,2-bis-(4-chloro-phenyl)-2-hydroxy-ethanone
show the reaction diagram
-
carboligation
17% yield and more than 99% enantiomeric excess of the (R)-enantiomer
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?
4-chlorobenzaldehyde + acetaldehyde
(S)-1-(4-Chloro-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
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?
4-Fluoro-benzaldehyde + acetaldehyde
(S)-1-(4-Fluoro-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
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-
-
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?
4-Fluoro-benzaldehyde + acetaldehyde
1-(4-Fluoro-phenyl)-2-hydroxy-propan-1-one
show the reaction diagram
-
87% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
4-fluorobenzaldehyde
(R)-1,2-Bis-(4-fluoro-phenyl)-2-hydroxy-ethanone
show the reaction diagram
-
carboligation
25% yield and more than 99% enantiomeric excess of the (R)-enantiomer
-
?
4-Formyl-benzonitrile + acetaldehyde
4-(2-Hydroxy-propionyl)-benzonitrile
show the reaction diagram
-
74% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
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?
4-formylbenzonitrile + acetaldehyde
4-((S)-2-Hydroxy-propionyl)-benzonitrile
show the reaction diagram
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-
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?
4-Hydroxy-benzaldehyde + acetaldehyde
2-Hydroxy-1-(4-hydroxy-phenyl)-propan-1-one
show the reaction diagram
-
86% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
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?
4-hydroxybenzaldehyde + acetaldehyde
(S)-2-Hydroxy-1-(4-hydroxy-phenyl)-propan-1-one
show the reaction diagram
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?
4-Methoxy-benzaldehyde + acetaldehyde
2-Hydroxy-1-(4-methoxy-phenyl)-propan-1-one
show the reaction diagram
-
92% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
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?
4-methoxybenzaldehyde
(R)-2-Hydroxy-1,2-bis-(4-methoxy-phenyl)-ethanone
show the reaction diagram
-
carboligation
12% yield and more than 99% enantiomeric excess of the (R)-enantiomer
-
?
4-methoxybenzaldehyde + acetaldehyde
(S)-2-Hydroxy-1-(4-methoxy-phenyl)-propan-1-one
show the reaction diagram
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-
-
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?
4-Methyl-benzaldehyde + acetaldehyde
2-Hydroxy-1-p-tolyl-propan-1-one
show the reaction diagram
-
88% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
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?
4-methylbenzaldehyde
(R)-2-Hydroxy-1,2-di-p-tolyl-ethanone
show the reaction diagram
-
carboligation
69% yield and more than 99% enantiomeric excess of the (R)-enantiomer
-
?
4-methylbenzaldehyde + acetaldehyde
(S)-2-Hydroxy-1-p-tolyl-propan-1-one
show the reaction diagram
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-
-
-
?
5-Isopropyl-furan-2-carbaldehyde + acetaldehyde
2-Hydroxy-1-(5-isopropyl-furan-2-yl)-propan-1-one
show the reaction diagram
-
73% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
5-methyl-furan-2-carbaldehyde
(R)-2-Hydroxy-1,2-bis-(5-methyl-furan-2-yl)-ethanone
show the reaction diagram
-
carboligation
50% yield and 96% enantiomeric excess of the (R)-enantiomer
-
?
5-Methyl-furan-2-carbaldehyde + acetaldehyde
2-Hydroxy-1-(5-methyl-furan-2-yl)-propan-1-one
show the reaction diagram
-
86% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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?
8-Hydroxy-quinoline-2-carbaldehyde + acetaldehyde
2-Hydroxy-1-(8-hydroxy-quinolin-2-yl)-propan-1-one
show the reaction diagram
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?
acetaldehyde
(R)-acetoin
show the reaction diagram
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?
acetaldehyde + acetaldehyde
(3R)-3-hydroxybutan-2-one
show the reaction diagram
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?
Acetic acid 3-formyl-phenyl ester + acetaldehyde
(S)-1-(3-Acetoxyphenyl)-2-hydroxy-propanone
show the reaction diagram
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-
-
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?
Acetic acid 3-formyl-phenyl ester + acetaldehyde
Acetic acid 3-(2-hydroxy-propionyl)-phenyl ester
show the reaction diagram
-
more than 99% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
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?
Benzaldehyde
(R)-Benzoin
show the reaction diagram
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benzaldehyde + acetaldehyde
(R)-2-hydroxypropiophenone
show the reaction diagram
-
reactions performed at high benzaldehyde concentrations and at high hydrostatic pressures show an increase in (R)-2-hydroxypropiophenone ((R)-2-HPP) formation catalyzed by BFD variants F464I, A460I, and A460I/F464I. For BFD mutant A460I/F464I there is an increase in the ee of (R)-2-HPP up to 80%, whereas at atmospheric conditions this variant synthesizes (R)-2-HPP with an ee of only 50%. Alkaline conditions (up to pH 8.5) and high hydrostatic pressures result in an increase of (R)-2-HPP synthesis, especially in the case of variant A460I and F464I
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?
benzaldehyde + acetaldehyde
(R)-benzoin + (S)-2-hydroxypropiophenone
show the reaction diagram
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-
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-
?
benzaldehyde + acetaldehyde
(S)-2-hydroxy-1-phenyl-propanone
show the reaction diagram
benzaldehyde + acetaldehyde
(S)-2-hydroxy-1-phenylpropan-1-one
show the reaction diagram
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-
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benzaldehyde + acetaldehyde
(S)-2-hydroxypropiophenone
show the reaction diagram
Benzaldehyde + acetaldehyde
2-Hydroxy-1-phenyl-propan-1-one
show the reaction diagram
-
92% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
benzaldehyde + acetaldehyde
2-hydroxypropiophenone
show the reaction diagram
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-
-
-
?
benzaldehyde + benzaldehyde
(2R)-2-hydroxy-1,2-diphenylethanone
show the reaction diagram
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?
benzaldehyde + benzaldehyde
(R)-benzoin
show the reaction diagram
Benzoylformate
(R)-Benzoin
show the reaction diagram
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Benzoylformate
?
show the reaction diagram
Benzoylformate
Benzaldehyde + CO2
show the reaction diagram
Benzoylformate + acetaldehyde
(S)-2-Hydroxypropiophenone
show the reaction diagram
benzylformate
benzaldehyde + CO2
show the reaction diagram
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-
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?
butyraldehyde
?
show the reaction diagram
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?
cyclohex-1-ene-1-carbaldehyde + acetaldehyde
(S)-1-Cyclohex-1-enyl-2-hydroxy-propan-1-one
show the reaction diagram
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?
Cyclohex-1-enecarbaldehyde + acetaldehyde
1-Cyclohex-1-enyl-2-hydroxy-propan-1-one
show the reaction diagram
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94% enantiomeric excess of the (S)-2-hydroxypropanone derivative
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?
cyclohexanecarbaldehyde + acetaldehyde
(S)-1-Cyclohexyl-2-hydroxy-propan-1-one
show the reaction diagram
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?
Cyclohexanecarbaldehyde + acetaldehyde
1-Cyclohexyl-2-hydroxy-propan-1-one
show the reaction diagram
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61% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
Furan-2-carbaldehyde + acetaldehyde
1-Furan-2-yl-2-hydroxy-propan-1-one
show the reaction diagram
-
45% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
isovaleraldehyde
(R)-5-hydroxy-2,7-dimethyloctan-4-one
show the reaction diagram
-
-
-
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?
m-bromo-benzoylformate
m-bromo-benzaldehyde + CO2
show the reaction diagram
-
wild-type enzyme: 68% conversion with 96% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme L476Q: 97% conversion with more than 98% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme M365L-L461S: 100% conversion with 98.5% enantiomeric excess of the (S)-2-hydroxy ketone
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?
m-chloro-benzoylformate
m-chloro-benzaldehyde + CO2
show the reaction diagram
-
wild-type enzyme: 94% conversion with 94% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme L476Q: 100% conversion with more than 97% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme M365L-L461S: 100% conversion with 98% enantiomeric excess of the (S)-2-hydroxy ketone
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?
m-fluoro-benzoylformate
m-fluoro-benzaldehyde + CO2
show the reaction diagram
-
wild-type enzyme: 100% conversion with 87% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme L476Q: 100% conversion with more than 99% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme M365L-L461S: 94% conversion with more than 99% enantiomeric excess of the (S)-2-hydroxy ketone
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?
m-methoxy-benzoylformate
m-methoxy-benzaldehyde + CO2
show the reaction diagram
-
wild-type enzyme: 94% conversion with 96% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme L476Q: 100% conversion with more than 99% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme M365L-L461S: 100% conversion with 99% enantiomeric excess of the (S)-2-hydroxy ketone
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?
m-methyl-benzoylformate
m-methyl-benzaldehyde + CO2
show the reaction diagram
-
wild-type enzyme: 99% conversion with 97% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme L476Q: 100% conversion with more than 99% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme M365L-L461S: 100% conversion with 99% enantiomeric excess of the (S)-2-hydroxy ketone
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-
?
methyl benzoylphosphonate
?
show the reaction diagram
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-
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?
n-butanal
(R)-5-hydroxyoctan-4-one
show the reaction diagram
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-
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-
?
n-pentanal
(R)-6-hydroxydecan-5-one
show the reaction diagram
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-
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-
?
Naphthalene-2-carbaldehyde + acetaldehyde
2-Hydroxy-1-naphthalen-2-yl-propan-1-one
show the reaction diagram
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-
-
-
?
nicotinaldehyde + acetaldehyde
(S)-2-Hydroxy-1-pyridin-3-yl-propan-1-one
show the reaction diagram
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-
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-
?
o-bromo-benzoylformate
o-bromo-benzaldehyde + CO2
show the reaction diagram
-
wild-type enzyme: no activity, mutant enzyme L476Q: 98% conversion with more than 99% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme M365L-L461S: 90% conversion with 99% enantiomeric excess of the (S)-2-hydroxy ketone
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-
?
o-chloro-benzoylformate
o-chloro-benzaldehyde + CO2
show the reaction diagram
-
wild-type enzyme: no activity, mutant enzyme L476Q: 100% conversion with more than 99% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme M365L-L461S: 85% conversion with more than 99% enantiomeric excess of the (S)-2-hydroxy ketone
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-
?
o-fluoro-benzoylformate
o-fluoro-benzaldehyde + CO2
show the reaction diagram
-
wild-type enzyme: 91% conversion with 89% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme L476Q: 100% conversion with more than 99% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme M365L-L461S: 83% conversion with 98% enantiomeric excess of the (S)-2-hydroxy ketone
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-
?
o-methoxy-benzoylformate
o-methoxy-benzaldehyde + CO2
show the reaction diagram
-
wild-type enzyme: no conversion, mutant enzyme L476Q: 97% conversion with more than 99% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme M365L-L461S: 46% conversion with 99% enantiomeric excess of the (S)-2-hydroxy ketone
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-
?
o-methyl-benzoylformate
o-methyl-benzaldehyde + CO2
show the reaction diagram
-
wild-type enzyme: 4% conversion, mutant enzyme L476Q: 100% conversion with more than 99% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme M365L-L461S: 83% conversion with 98% enantiomeric excess of the (S)-2-hydroxy ketone
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-
?
o-methylbenzaldehyde + acetaldehyde
(2S)-2-hydroxy-1-(2-methylphenyl)propan-1-one
show the reaction diagram
-
mutant enzyme L476Q and M365L/L461S selectively catalyzes the formation of enantiopure (S)-2-hydroxy-1-(2-methylphenyl)propan-1-one with excellent yield, a reaction which is only poorly catalyzed by the wild-type enzyme
-
-
?
p-bromo-benzoylformate
p-bromo-benzaldehyde + CO2
show the reaction diagram
-
wild-type enzyme: 42% conversion with 83% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme L476Q: 100% conversion with 96.5% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme M365L-L461S: 100% conversion with 96% enantiomeric excess of the (S)-2-hydroxy ketone
-
-
?
p-chloro-benzoylformate
p-chloro-benzaldehyde + CO2
show the reaction diagram
-
wild-type enzyme: 85% conversion with 82% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme L476Q: 100% conversion with more than 96.5% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme M365L-L461S: 100% conversion with 95.5% enantiomeric excess of the (S)-2-hydroxy ketone
-
-
?
p-fluoro-benzoylformate
p-fluoro-benzaldehyde + CO2
show the reaction diagram
-
wild-type enzyme: 69% conversion with 87% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme L476Q: 100% conversion with 97% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme M365L-L461S: 87% conversion with 97% enantiomeric excess of the (S)-2-hydroxy ketone
-
-
?
p-methoxy-benzoylformate
p-methoxy-benzaldehyde + CO2
show the reaction diagram
-
wild-type enzyme: 23% conversion with 92% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme L476Q: 100% conversion with more than 99% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme M365L-L461S: 67% conversion with 42% enantiomeric excess of the (S)-2-hydroxy ketone
-
-
?
p-methyl-benzoylformate
p-methyl-benzaldehyde + CO2
show the reaction diagram
-
wild-type enzyme: 65% conversion with 88% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme L476Q: 100% conversion with more than 98% enantiomeric excess of the (S)-2-hydroxy ketone, mutant enzyme M365L-L461S: 98% conversion with 98% enantiomeric excess of the (S)-2-hydroxy ketone
-
-
?
p-nitro-benzoylformate
?
show the reaction diagram
-
very poor substrate
-
-
?
p-nitrobenzoylformate
p-nitrobenzaldehyde + CO2
show the reaction diagram
-
-
-
-
?
phenylglyoxylate
benzaldehyde + CO2
show the reaction diagram
propanal
4-hydroxyhexan-3-one
show the reaction diagram
-
-
-
-
?
propionaldehyde
?
show the reaction diagram
-
-
-
-
?
pyridine-2-carbaldehyde
(R)-2-Hydroxy-1,2-di-pyridin-2-yl-ethanone
show the reaction diagram
-
carboligation
70% yield and 94% enantiomeric excess of the (R)-enantiomer
-
?
Pyridine-2-carbaldehyde + acetaldehyde
2-Hydroxy-1-pyridin-2-yl-propan-1-one
show the reaction diagram
-
-
-
-
?
Pyridine-3-carbaldehyde + acetaldehyde
2-Hydroxy-1-pyridin-3-yl-propan-1-one
show the reaction diagram
-
-
-
-
?
pyruvate
acetaldehyde + CO2
show the reaction diagram
Quinoline-4-carbaldehyde + acetaldehyde
2-Hydroxy-1-quinolin-4-yl-propan-1-one
show the reaction diagram
-
-
-
-
?
thiophene-2-carbaldehyde
(S)-2-Hydroxy-1,2-di-thiophen-2-yl-ethanone
show the reaction diagram
-
carboligation
65% yield and 95% enantiomeric excess of the (R)-enantiomer
-
?
thiophene-2-carbaldehyde + acetaldehyde
(S)-2-Hydroxy-1-thiophen-2-yl-propan-1-one
show the reaction diagram
-
-
-
-
?
Thiophene-2-carbaldehyde + acetaldehyde
2-Hydroxy-1-thiophen-2-yl-propan-1-one
show the reaction diagram
-
83% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
[m-(Fluoromethyl)benzoyl]formate
?
show the reaction diagram
-
-
-
-
-
[p-(Chloromethyl)benzoyl]formate
?
show the reaction diagram
-
-
-
-
-
[p-(Fluoromethyl)benzoyl]formate
?
show the reaction diagram
-
-
-
-
-
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
3-ethoxy-4-hydroxybenzoylformate
ethyl vanillin + CO2
show the reaction diagram
Benzoylformate
?
show the reaction diagram
Benzoylformate
Benzaldehyde + CO2
show the reaction diagram
-
-
-
-
?
phenylglyoxylate
benzaldehyde + CO2
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
thiamine diphosphate
additional information
-
the addition of thiamine thiazolone diphosphate has no effect on the activity of BFD in the presence of thiamine diphosphate
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
bound to the cofactor in each subunit
Zn2+
-
activates
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(R)-mandelate
(S)-Mandelate
-
competitive
2-carboxybenzaldehyde
-
competitive
2-oxo-4-methylhexanoic acid
-
substrate inhibition
2-oxohexanoic acid
-
substrate inhibition
2-Oxopentanoic acid
-
substrate inhibition
4-chloromercuribenzoate
-
-
acetylphosphonate
-
-
benzoylformate
benzoylphosphonate
citrate
-
-
methyl acetylphosphonate
-
-
methyl benzoylphosphonate
Phenoxyacetate
-
competitive
phenylacetate
-
competitive
Phthalate
-
competitive
Phthalimide
-
competitive
R-mandelate
-
a substrate-analogue inhibitor, the carboxylate of the inhibitor is in an apparent hydrogen bond with Ser26, whereas one of the histidine residues, His70, is positioned to act as a proton donor in the formation of the first tetrahedral intermediate, mandelyl-ThDP
trans-cinnamate
-
competitive
[p-(Bromomethyl)benzoyl]formate
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
thiamine diphosphate
-
half-maximal activity below 0.001 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.056
(p-methoxybenzoyl)formate
-
-
0.19 - 0.58
(p-methylbenzoyl)formate
0.42 - 7.5
2-ketobutanoate
0.15 - 4.1
2-ketohexanoate
0.1 - 6
2-ketopentanoate
1.1
2-oxo-4-methylhexanoic acid
-
pH 6.0, 30C, mutant enzyme A460I
3.4 - 9.6
2-oxo-4-methylpentanoic acid
18.3
2-oxobutanoic acid
-
pH 6.0, 30C, mutant enzyme A460I
1 - 4.1
2-oxohexanoic acid
1.2 - 6
2-Oxopentanoic acid
310 - 775
acetaldehyde
19.4 - 77
benzaldehyde
0.059 - 8.62
benzoylformate
0.04 - 1.5
benzylformate
0.76
methyl benzoylphosphonate
-
-
0.15
p-nitro-benzoylformate
-
at 25C
0.27 - 7.8
phenylglyoxylate
2 - 21
pyruvate
0.023
[m-(fluoromethyl)benzoyl]formate
-
-
-
0.021 - 0.18
[p-(Chloromethyl)benzoyl]formate
0.19
[p-(Fluoromethyl)benzoyl]formate
-
-
additional information
additional information
-
KM-value of wild-type enzyme for 2-oxo-4-methylhexanoic acid (pH 6.0, 30C) is above 20 mM
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4 - 7.5
2-ketobutanoate
1.1 - 5.3
2-ketohexanoate
0.81 - 11
2-ketopentanoate
8.6 - 9.8
2-oxo-4-methylhexanoic acid
11 - 38
2-oxo-4-methylpentanoic acid
17.3 - 18.8
2-oxobutanoic acid
21 - 71.8
2-oxohexanoic acid
40.6 - 44.3
2-Oxopentanoic acid
0.07 - 1349
benzoylformate
1.7 - 320
benzylformate
0.05
p-nitro-benzoylformate
Pseudomonas putida
-
at 25C
0.46 - 320
phenylglyoxylate
1.4 - 3.6
pyruvate
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.5 - 14
2-ketobutanoate
3288
0.86 - 16
2-ketohexanoate
4407
1.8 - 22
2-ketopentanoate
3158
16 - 1180
benzylformate
7158
0.3 - 1180
phenylglyoxylate
4044
0.012 - 1.8
pyruvate
31
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1 - 100
(R)-mandelate
48
2-oxo-4-methylhexanoic acid
-
pH 6.0, 30C, mutant enzyme A460I
2 - 10
2-oxohexanoic acid
20
2-Oxopentanoic acid
-
pH 6.0, 30C, wild-type enzyme and mutant enzyme A460I
45 - 143
benzoylformate
0.36
benzoylphosphonate
-
prolonged incubation of BFD with benzoylphosphonate results in permanent inactivation of the enzyme
0.38
methyl benzoylphosphonate
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
11
-
activity of free BFD
additional information
-
After 24 h of catalytic activity the immobilized biocatalyst Sepabeads EC-EA/BFD showed a remaining activity of 5% of its initial value.; specific activity of around 62 U/g dry carrier. The activity is determined for the reaction of acetaldehyde (500 mM) and benzaldehyde (50 mM) at pH 8.0; Stability of Sepabeads EC-EA/BFD is determined by repetitive batch experiments while carrying out the synthesis of 2-hydroxy-1-phenylpropanone. The reaction of benzaldehyde and acetaldehyde is carried out at 30C and after each batch is washed. The decrease of the initial reaction rate is followed for six batches. The newly prepared biocatalyst (enzyme loading 320 mg/g) shows an initial reaction rate of 130 U/g carrier with respect to the C-C-coupling reaction, whereby the rate decreases down to 10% (13 U/g) in the sixth batch.
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
-
assay at, substrate phenylglyoxylate
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 8
-
pH 4.0: about 50% of maximal activity, pH 8.0: about 60% of maximal activity, decaboxylation of benzoylformate
5 - 8.5
-
pH 5.0: about 60% of maximal activity, pH 8.5: about 80% of maximal activity, carboligation with benzoylformate and acetaldehyde as substrate
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20
-
assay at
45
-
temperature optimum for immobilized variant A460I/F464I
50
-
at pH 8.0
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 70
-
At temperatures above 70C and lower than 20C the activity decreases below 10 U/g carrier.
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
no activity with benzoate or glucose as growth substrate
Manually annotated by BRENDA team
-
no activity with benzoate or glucose as growth substrate
Manually annotated by BRENDA team
-
no activity with benzoate or glucose as growth substrate
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)
Polynucleobacter necessarius subsp. asymbioticus (strain DSM 18221 / CIP 109841 / QLW-P1DMWA-1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
53614
-
4 * 53614, calculation from nucleotide sequence
55000
-
4 * 55000, gel filtration
56000
-
x * 56000, recombinant His-tagged enzyme, SDS-PAGE
57000
-
4 * 57000
80000
-
gel filtration
220000
-
gel filtration
248000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 56000, recombinant His-tagged enzyme, SDS-PAGE
homotetramer
tetramer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, structure of the enzyme complexed with the inhibitor (R)-mandelate
-
in complex with (E)-2-oxo-4(pyridin-3-yl)-3-butenoic acid or 3-(pyridin-3-yl)acrylaldehyde, hanging drop vapour diffusion method, in 100 mM TrisHCl (pH 8.5), 150 mM CaCl2, 0.5% (v/v) MPD [2-methyl-2,4-pentanediol], and 22% (v/v) PEG 400
-
in complex with methyl benzoylphosphonate, hanging drop vapor diffusion method, using 100 mM Tris-HCl (pH 8.5), 150 mM CaCl2, 22% (v/v) PEG 400
-
mutant enzyme L461A, hanging drop vapour diffusion method
-
sitting drop vapor diffusion method, using 25% (w/v) polyethylene glycol 2000 and 0.2 M MgCl2, at pH 6.
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3
-
deactivation is exponential
655230
5.5 - 7.5
-
stable
4811
5.5
-
deactivation kinetics is almost linear with 9.3% per min
655230
10
-
deactivation 0.1% per min
655230
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
BFD is stabilized by increasing water activity
-
significant stabilizing effect of thiamin diphosphate and Mg2+
-
the activity of the spherical silica-immobilized BFD A460I-F464I variant is determined to be 70% related to the activity of the free enzyme
-
The freeze-drying of immobilized Sepabeads EC-EA/BFD leads to an inactivation of the enzyme.
-
the wild type BFD has about 100% enzyme activity after 24 h incubation in 50 mM KPi-buffer without thiamine diphosphate
-
the wild type enzyme shows no loss of activity in cofactor-free buffer over 24 h
-
urea, less than 4 M, reversibly inactivates, thiamin diphosphate protects
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
DMSO
-
retains its full activity after 10 days of incubation in DMSO (20% v/v) at 25C
Ethanol
-
retains its full activity after 10 days of incubation in aqueous ethanol (1.5 M) at 25C
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-15C, stable for 48 h
-
-20C, lyophilized His-tagged enzyme is stable for several months
-
20C, pH 7, in absence of cofactor and substrates, half-life is 3 days
-
20C, pH 7, in presence of 0.5 mM thiamin diphosphate and 0.5 mM Mg2+, half-life is 36 d
-
20C, pH 7, in presence of 0.5 mM thiamin diphosphate, 100 mM acetaldehyde and 0.5 mM Mg2+, half-life is 1.8 d
-
20C, pH 7, in presence of 0.5 mM thiamin diphosphate, 200 mM acetaldehyde and 0.5 mM Mg2+, half-life is 1.5 d
-
20C, pH 7, in presence of 0.5 mM thiamin diphosphate, 25 mM benzaldehyde and 0.5 mM Mg2+, half-life is 5.3 d
-
4C, pH 7, in presence of 0.5 mM thiamin diphosphate and 0.5 mM Mg2+, half-life is 139 d
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by Ni-NTA column purification.
-
large-scale
-
Ni-NTA column chromatography
-
Ni-NTA column chromatography and Superdex G200 gel filtration
-
Phenyl Sepharose FF column chromatography and Q Sepharose FF column chromatography
-
Q-Sepharose column chromatography and Superdex 200 gel filtration
-
Q-Sepharose column chromatography, Superdex 200 gel filtration
-
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography
-
recombinant His-tagged enzyme from Escherichia coli strain SG13009 by nickel affinity chromatography
-
recombinant wild-type and mutant enzymes
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli BL21(DE3)pLysS cells
-
expressed in Escherichia coli Escherichia coli SG13009 pKK233-2
-
expressed in Escherichia coli SG13009 cells
-
expressed in Escherichia coli strain BL21(DE3)
-
expressed in Escherichia coli strain SG13009
-
expressed in Escherichia coli strain SG13009/pREP4
-
expression as a hexahistidine fusion protein in Escherichia coli
-
expression of His-tagged enzyme in Escherichia coli strain SG13009
-
expression of mutant enzyme L476Q and M365L/L461S in Escherichia coli
-
gene mdlC, recombinant expression in Escherichia coli strain BL21(DE3), suncloning in Escherichia coli strain DH5alpha
Production as hexahistidine fusion protein by fermentation of Escherichia coli cells as host strain.
-
recombinant expression of His-tagged enzyme in Escherichia coli strain SG13009
-
wild-type and mutant enzymes A460I, F464I and A460I/F464I
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A460G
-
mutant exhibits a higher Km value compared to the wild type enzyme
A460I/F464I
G464I
-
activity with pyruvate is higher than wild-type activity, activity with 2-oxobutanoic acid is 1.2fold lower than wild-type activity, activity with 2-oxopentanoic acid is 4.8fold lower than wild-type value, activity with 2-oxohexanoic acid is 1.6fold lower than wild-type activity, activity with 3-methyl-2-oxobutanoic acid is 1.1fold higher than wild-type activity, activity with 3-methyl-2-oxopentanoic acid is 1.3fold higher than wild-type activity, activity with 2-oxo-4-methylpentanoic acid is 1.1fold higher than wild-type activity, activity with 4,4-dimethyl-2-oxopentanoic acid is identical to wild-type activity, activity with 2-oxo-4-methylhexanoic acid is 1.3fold higher than wild-type activity, activity with, activity with benzoylformate is 9.2fold lower than wild-type activity, activity with 2-oxo-3-phenylpropanoic acid is identical to wild-type activity, activity with 2-oxo-4-phenylbutanoic acid is9.3fold lower than wild-type activity, no activity with 2-oxo-5-phenylpentanoic acid. The ratio of turnover number to KM-value for benzoylformate is 31.8fold lower than the wild-type value. Mutation has no effect on the range of products obtained by carboligation of acetaldehyde and benzaldehyde, the yield of the product 2-hydroxypropiophenone an decreases about 3fold and the enantioselectivity of acetoin and 2-hydroxypropiophenone is altered
H281N
-
mutant with 17fold decrease in kcat value compared to the wild type enzyme
H281Q
-
mutant with 37fold decrease in kcat value compared to the wild type enzyme
H281T
-
mutant with 159fold decrease in kcat value compared to the wild type enzyme
H281W
-
mutant with 19fold decrease in kcat value compared to the wild type enzyme
H70S
-
mutant exhibits a 197fold decrease in kcat/Km compared to the wild type enzyme
L403E
-
kcat value of the L403E variant is only 18fold lower than that of wild-type BFDC
L403F
-
L403F variant shows about 10fold increased Km value for the substrate compared to the wild-type enzyme, the cofactor is displaced with the thiazolium ring away
L403X
-
half the L403X colonies screened have at least 10% of wild-type activity
L461V
-
mutant exhibits a higher Km value compared to the wild type enzyme
L476A
-
has a 4.11fold higher carboligase activity than the wild type enzyme
L476C
-
has a 4.31fold higher carboligase activity than the wild type enzyme
L476G
-
has a 4.27fold higher carboligase activity than the wild type enzyme
L476K
-
has a 5fold higher carboligase activity than the wild type enzyme
L476M
-
has a 4.23fold higher carboligase activity than the wild type enzyme
L476P/S181T
-
mutant with low cofactor binding affinity
L476Q/S535G
-
decreased activity
L476S
-
has a 3.5fold higher carboligase activity than the wild type enzyme
L476T
-
has a 3.98fold higher carboligase activity than the wild type enzyme
M365L/L461S
-
mutant enzyme selectively catalyzes the formation of enantiopure (S)-2-hydroxy-1-(2-methylphenyl)propan-1-one with excellent yield, a reaction which is only poorly catalyzed by the wild-type enzyme. Mutant enzyme retains only 9% of the wild-type BFD carboligase activity. Decrease in Vmax value as well as an increase in the Km-value for decarboxylation of benzoylformate by mutant enzyme compared to that of wild-type enzyme
P24A
-
mutant exhibits a higher Km value compared to the wild type enzyme
S181T/L476P
-
increased activity
S26T
-
mutant exhibits no significant loss of activity compared to the wild type enzyme (3fold decrease in kcat value)
T377L/A460Y
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
-
Production of immobilized enzyme on Sepabeads EC-EA by C-C coupling
synthesis
Show AA Sequence (394 entries)
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