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Enzyme Nomenclature
Information on EC 4.1.1.7 - benzoylformate decarboxylase
for references in articles please use BRENDA:EC4.1.1.7
Word Map on EC 4.1.1.7
- 4.1.1.7
- thiamin
- putida
-
diphosphate-dependent
-
thdp-dependent
- acetaldehyde
-
carboligation
-
decarboxylases
-
r-benzoin
- r-mandelate
- acyloin
-
2-keto
-
carboligase
-
2-hydroxy
- synthesis
- biotechnology
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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
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EC NUMBER 
COMMENTARY 
4.1.1.7
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RECOMMENDED NAME
GeneOntology No.
benzoylformate decarboxylase
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
phenylglyoxylate = benzaldehyde + CO2
the reaction has two partially rate-determining steps: initial tetrahedral adduct formation and decarboxylation
-
phenylglyoxylate = benzaldehyde + CO2
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-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
decarboxylation
decarboxylation
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-
-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
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SYSTEMATIC NAME
IUBMB Comments
benzoylformate carboxy-lyase (benzaldehyde-forming)
A thiamine-diphosphate protein.
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CAS REGISTRY NUMBER
COMMENTARY
9025-00-7
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
no antigenic cross-reactivity with the enzymes of Pseudomonas aeruginosa and Pseudomonas putida
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-
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4799, 4800, 4801, 4802, 4803, 4807, 4808, 4810, 4811, 4813, 4814, 655230, 655499, 657382, 677679, 678222, 678577, 679212, 679656, 680352, 680979, 682628, 691346, 702796, 715697, 716121, 727333, 727483, 728161
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
-
penultimate enzyme in the mandelate pathway, that catalyzes the non-oxidative decarboxylation of benzoylformate to yield carbon dioxide and benzaldehyde
physiological function
additional information
-
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
physiological function
-
the enzyme functions in decarboxylation of benzoylformate to benzaldehyde in the mandalate pathway
physiological function
-
the enzyme enables the organism to utilize R-mandelate as their sole carbon source
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(E)-2-oxo-4(pyridin-3-yl)-3-butenoic acid
3-(pyridin-3-yl)acrylaldehyde + CO2
-
-
-
-
?
2 benzaldehyde
(R)-benzoin
-
-
i.e. (R)-2-hydroxy-1,2-diphenylethan-1-one
-
?
2-Fluoro-benzaldehyde + acetaldehyde
(S)-1-(2-Fluoro-phenyl)-2-hydroxy-propan-1-one
-
-
-
-
?
2-Fluoro-benzaldehyde + acetaldehyde
1-(2-Fluoro-phenyl)-2-hydroxy-propan-1-one
-
91% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
2-fluorobenzaldehyde
(R)-1,2-Bis-(2-fluoro-phenyl)-2-hydroxy-ethanone
-
carboligation
68% yield and more than 99% enantiomeric excess of the (R)-enantiomer
-
?
2-furaldehyde
(R)-1,2-di-furan-2-yl-2-hydroxy-ethanone
-
carboligation
62% yield and 94% enantiomeric excess of the (R)-enantiomer
-
?
2-furaldehyde + acetaldehyde
(S)-1-Furan-2-yl-2-hydroxy-propan-1-one
-
-
-
-
?
2-Methyl-benzaldehyde + acetaldehyde
(S)-2-Hydroxy-1-(2-methylphenyl)-propanone
-
-
-
-
?
2-Methyl-benzaldehyde + acetaldehyde
2-Hydroxy-1-o-tolyl-propan-1-one
-
4% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
3,5-Difluoro-benzaldehyde + acetaldehyde
(S)-1-(3,5-Difluoro-phenyl)-2-hydroxy-propan-1-one
-
-
-
-
?
3,5-dimethoxybenzaldehyde + acetaldehyde
(S)-1-(3,5-Dimethoxy-phenyl)-2-hydroxy-propan-1-one
-
-
-
-
?
3-Benzyloxy-benzaldehyde + acetaldehyde
1-(3-Benzyloxy-phenyl)-2-hydroxy-propan-1-one
-
more than 99% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
3-Bromo-benzaldehyde + acetaldehyde
(S)-1-(3-Bromo-phenyl)-2-hydroxy-propan-1-one
-
-
-
-
?
3-Bromo-benzaldehyde + acetaldehyde
1-(3-Bromo-phenyl)-2-hydroxy-propan-1-one
-
96% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
3-Chloro-benzaldehyde + acetaldehyde
(S)-1-(3-Chloro-phenyl)-2-hydroxy-propan-1-one
-
-
-
-
?
3-Chloro-benzaldehyde + acetaldehyde
1-(3-Chloro-phenyl)-2-hydroxy-propan-1-one
-
94% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
3-Ethoxy-benzaldehyde + acetaldehyde
(S)-1-(3-Ethoxy-phenyl)-2-hydroxy-propan-1-one
-
-
-
-
?
3-Ethoxy-benzaldehyde + acetaldehyde
1-(3-Ethoxy-phenyl)-2-hydroxy-propan-1-one
-
97% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
3-Fluoro-benzaldehyde + acetaldehyde
(S)-1-(3-Fluoro-phenyl)-2-hydroxy-propan-1-one
-
-
-
-
?
3-Fluoro-benzaldehyde + acetaldehyde
1-(3-Fluoro-phenyl)-2-hydroxy-propan-1-one
-
87% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
3-Formyl-benzonitrile + acetaldehyde
3-((S)-2-Hydroxy-propionyl)-benzonitrile
-
-
-
-
?
3-Formyl-benzonitrile + acetaldehyde
3-(2-Hydroxy-propionyl)-benzonitrile
-
92% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
3-Hydroxy-benzaldehyde + acetaldehyde
(S)-2-Hydroxy-1-(3-hydroxy-phenyl)-propan-1-one
-
-
-
-
?
3-Hydroxy-benzaldehyde + acetaldehyde
2-Hydroxy-1-(3-hydroxy-phenyl)-propan-1-one
-
92% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
3-Isopropoxy-benzaldehyde + acetaldehyde
(S)-2-Hydroxy-1-(3-isopropoxy-phenyl)-propan-1-one
-
-
-
-
?
3-Isopropoxy-benzaldehyde + acetaldehyde
2-Hydroxy-1-(3-isopropoxy-phenyl)-propan-1-one
-
more thahn 99% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
3-Methoxy-benzaldehyde + acetaldehyde
(S)-2-Hydroxy-1-(3-methoxy-phenyl)-propan-1-one
-
-
-
-
?
3-Methoxy-benzaldehyde + acetaldehyde
2-Hydroxy-1-(3-methoxy-phenyl)-propan-1-one
-
96% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
3-methoxybenzaldehyde
(R)-2-Hydroxy-1,2-bis-(3-methoxy-phenyl)-ethanone
-
carboligation
18% yield and more than 99% enantiomeric excess of the (R)-enantiomer
-
?
3-Methoxymethoxy-benzaldehyde + acetaldehyde
(S)-2-Hydroxy-1-(3-methoxymethoxy-phenyl)-propan-1-one
-
-
-
-
?
3-Methyl-benzaldehyde + acetaldehyde
(S)-2-Hydroxy-1-m-tolyl-propan-1-one
-
-
-
-
?
3-Methyl-benzaldehyde + acetaldehyde
2-Hydroxy-1-m-tolyl-propan-1-one
-
97% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
3-Phenoxy-benzaldehyde + acetaldehyde
2-Hydroxy-1-(3-phenoxy-phenyl)-propan-1-one
-
more than 99% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
4-Bromo-benzaldehyde + acetaldehyde
1-(4-Bromo-phenyl)-2-hydroxy-propan-1-one
-
83% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
4-bromobenzaldehyde
(R)-1,2-bis-(4-bromo-phenyl)-2-hydroxy-ethanone
-
carboligation
13% yield and more than 99% enantiomeric excess of the (R)-enantiomer
-
?
4-bromobenzaldehyde + acetaldehyde
(S)-1-(4-Bromo-phenyl)-2-hydroxy-propan-1-one
-
-
-
-
?
4-Chloro-benzaldehyde + acetaldehyde
1-(4-Chloro-phenyl)-2-hydroxy-propan-1-one
-
82% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
4-chlorobenzaldehyde
(R)-1,2-bis-(4-chloro-phenyl)-2-hydroxy-ethanone
-
carboligation
17% yield and more than 99% enantiomeric excess of the (R)-enantiomer
-
?
4-chlorobenzaldehyde + acetaldehyde
(S)-1-(4-Chloro-phenyl)-2-hydroxy-propan-1-one
-
-
-
-
?
4-Fluoro-benzaldehyde + acetaldehyde
(S)-1-(4-Fluoro-phenyl)-2-hydroxy-propan-1-one
-
-
-
-
?
4-Fluoro-benzaldehyde + acetaldehyde
1-(4-Fluoro-phenyl)-2-hydroxy-propan-1-one
-
87% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
4-fluorobenzaldehyde
(R)-1,2-Bis-(4-fluoro-phenyl)-2-hydroxy-ethanone
-
carboligation
25% yield and more than 99% enantiomeric excess of the (R)-enantiomer
-
?
4-Formyl-benzonitrile + acetaldehyde
4-(2-Hydroxy-propionyl)-benzonitrile
-
74% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
4-formylbenzonitrile + acetaldehyde
4-((S)-2-Hydroxy-propionyl)-benzonitrile
-
-
-
-
?
4-Hydroxy-benzaldehyde + acetaldehyde
2-Hydroxy-1-(4-hydroxy-phenyl)-propan-1-one
-
86% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
4-hydroxybenzaldehyde + acetaldehyde
(S)-2-Hydroxy-1-(4-hydroxy-phenyl)-propan-1-one
-
-
-
-
?
4-Methoxy-benzaldehyde + acetaldehyde
2-Hydroxy-1-(4-methoxy-phenyl)-propan-1-one
-
92% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
4-methoxybenzaldehyde
(R)-2-Hydroxy-1,2-bis-(4-methoxy-phenyl)-ethanone
-
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
-
-
-
-
?
4-Methyl-benzaldehyde + acetaldehyde
2-Hydroxy-1-p-tolyl-propan-1-one
-
88% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
4-methylbenzaldehyde
(R)-2-Hydroxy-1,2-di-p-tolyl-ethanone
-
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
-
-
-
-
?
5-Isopropyl-furan-2-carbaldehyde + acetaldehyde
2-Hydroxy-1-(5-isopropyl-furan-2-yl)-propan-1-one
-
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
-
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
-
86% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
8-Hydroxy-quinoline-2-carbaldehyde + acetaldehyde
2-Hydroxy-1-(8-hydroxy-quinolin-2-yl)-propan-1-one
-
-
-
-
?
Acetic acid 3-formyl-phenyl ester + acetaldehyde
(S)-1-(3-Acetoxyphenyl)-2-hydroxy-propanone
-
-
-
-
?
Acetic acid 3-formyl-phenyl ester + acetaldehyde
Acetic acid 3-(2-hydroxy-propionyl)-phenyl ester
-
more than 99% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
benzaldehyde + acetaldehyde
(R)-2-hydroxypropiophenone
-
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
-
-
?
benzaldehyde + acetaldehyde
(R)-benzoin + (S)-2-hydroxypropiophenone
-
-
-
-
?
Benzaldehyde + acetaldehyde
2-Hydroxy-1-phenyl-propan-1-one
-
92% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
cyclohex-1-ene-1-carbaldehyde + acetaldehyde
(S)-1-Cyclohex-1-enyl-2-hydroxy-propan-1-one
-
-
-
-
?
Cyclohex-1-enecarbaldehyde + acetaldehyde
1-Cyclohex-1-enyl-2-hydroxy-propan-1-one
-
94% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
cyclohexanecarbaldehyde + acetaldehyde
(S)-1-Cyclohexyl-2-hydroxy-propan-1-one
-
-
-
-
?
Cyclohexanecarbaldehyde + acetaldehyde
1-Cyclohexyl-2-hydroxy-propan-1-one
-
61% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
Furan-2-carbaldehyde + acetaldehyde
1-Furan-2-yl-2-hydroxy-propan-1-one
-
45% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
m-bromo-benzoylformate
m-bromo-benzaldehyde + CO2
-
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
-
-
?
m-chloro-benzoylformate
m-chloro-benzaldehyde + CO2
-
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
-
-
?
m-fluoro-benzoylformate
m-fluoro-benzaldehyde + CO2
-
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
-
-
?
m-methoxy-benzoylformate
m-methoxy-benzaldehyde + CO2
-
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
-
-
?
m-methyl-benzoylformate
m-methyl-benzaldehyde + CO2
-
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
-
-
?
Naphthalene-2-carbaldehyde + acetaldehyde
2-Hydroxy-1-naphthalen-2-yl-propan-1-one
-
-
-
-
?
nicotinaldehyde + acetaldehyde
(S)-2-Hydroxy-1-pyridin-3-yl-propan-1-one
-
-
-
-
?
o-bromo-benzoylformate
o-bromo-benzaldehyde + CO2
-
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
-
-
?
o-chloro-benzoylformate
o-chloro-benzaldehyde + CO2
-
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
-
-
?
o-fluoro-benzoylformate
o-fluoro-benzaldehyde + CO2
-
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
-
-
?
o-methoxy-benzoylformate
o-methoxy-benzaldehyde + CO2
-
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
-
-
?
o-methyl-benzoylformate
o-methyl-benzaldehyde + CO2
-
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
-
-
?
o-methylbenzaldehyde + acetaldehyde
(2S)-2-hydroxy-1-(2-methylphenyl)propan-1-one
-
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
-
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
-
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
-
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
-
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
-
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
-
-
?
pyridine-2-carbaldehyde
(R)-2-Hydroxy-1,2-di-pyridin-2-yl-ethanone
-
carboligation
70% yield and 94% enantiomeric excess of the (R)-enantiomer
-
?
Pyridine-2-carbaldehyde + acetaldehyde
2-Hydroxy-1-pyridin-2-yl-propan-1-one
-
-
-
-
?
Pyridine-3-carbaldehyde + acetaldehyde
2-Hydroxy-1-pyridin-3-yl-propan-1-one
-
-
-
-
?
Quinoline-4-carbaldehyde + acetaldehyde
2-Hydroxy-1-quinolin-4-yl-propan-1-one
-
-
-
-
?
thiophene-2-carbaldehyde
(S)-2-Hydroxy-1,2-di-thiophen-2-yl-ethanone
-
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
-
-
-
-
?
Thiophene-2-carbaldehyde + acetaldehyde
2-Hydroxy-1-thiophen-2-yl-propan-1-one
-
83% enantiomeric excess of the (S)-2-hydroxypropanone derivative
-
-
?
3-ethoxy-4-hydroxybenzoylformate
ethyl vanillin + CO2
-
-
-
?
benzaldehyde + acetaldehyde
(S)-2-hydroxy-1-phenyl-propanone
-
the enantiomeric excess of (S)-2-hydroxy-1-phenyl-propanonedecreases with increasing temperature from 4°C to 30°C
-
-
?
benzaldehyde + acetaldehyde
(S)-2-hydroxypropiophenone
-
-
i.e. acyloin
-
?
benzaldehyde + benzaldehyde
(R)-benzoin
-
carboligation
70% yield and more than 99% enantiomeric excess of the (R)-enantiomer
-
?
Benzoylformate
?
-
enzyme is involved in the inducible pathway of mandelate
-
-
-
Benzoylformate
?
-
enzyme is involved in the metabolism of mandelic acid
-
-
-
Benzoylformate
?
-
enzyme is involved in the metabolism of mandelic acid
-
-
-
Benzoylformate
?
-
enzyme is involved in the inducible pathway of mandelate
-
-
-
Benzoylformate
?
-
enzyme is involved in the metabolism of mandelic acid
-
-
-
Benzoylformate
?
-
enzyme is involved in the metabolic pathway of phenylglycine
-
-
-
Benzoylformate
?
-
enzyme is involved in the metabolic pathway of phenylglycine
-
-
-
Benzoylformate
?
-
the enzyme is involved in the pathway for catabolism of D(+)-mandelate and L-(-)-mandelate
-
-
-
Benzoylformate
Benzaldehyde + CO2
-
-
654692, 655230, 655240, 677679, 678222, 678577, 679209, 679212, 679656, 680352, 680368, 680979, 682628, 691794, 715697, 716121
-
-
?
Benzoylformate
Benzaldehyde + CO2
-
best substrate for wild-type enzyme and mutant enzymes A460I, F464I, and A4670I, F464I
-
-
?
Benzoylformate + acetaldehyde
(S)-2-Hydroxypropiophenone
-
-
benzaldehyde and benzyl alcohol are formed as by-products
-
pyruvate
acetaldehyde + CO2
-
activity of wild-type enzyme is very low, very low activity with mutant enzymes A460I, F464I, and A4670I, F464I
-
-
?
pyruvate
acetaldehyde + CO2
-
pyruvate is no substrate for the wild type enzyme
-
-
?
additional information
?
-
-
best substrates in enantioselective synthesis of (S)-2-hydroxypropanone derivatives by C-C bond formation are meta-substituted benzaldehyde derivatives
-
-
-
additional information
?
-
-
no activity with 3,3-dimethyl-2-oxopentanoic acid and cyclohexaneglyoxylic acid (wild-type enzyme and mutant enzymes A460I, F464I and A460I/F464I)
-
-
-
additional information
?
-
-
does not perform the cleavage of 2-hydroxy ketones
-
-
-
additional information
?
-
-
isobutyraldehyde, pivalaldehyde, and tert-butylacetaldehyde are no substrates
-
-
-
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
-
-
-
additional information
?
-
-
the enzyme performs enantioselective synthesis of (S)-2-hydroxyketones. Its stereoselectivity is highly dependent on the structure of the substrate aldehydes
-
-
-
additional information
?
-
when 3-ethoxy-4-hydroxybenzoylformate is provided at with Pseudomonas putida, it is completely transformed to ethyl vanillic acid after 2 days, but only trace amounts of ethyl vanillin are detected. Similar results are obtained when ethyl vanillin replaces 3-ethoxy-4-hydroxybenzoylformate as substrate, thus aldehyde dehydrogenation activity in Pseudomonas putida is greater than its synthesis activity. The recombinant Escherichia coli strain expressing the enzyme from Pseudomonas putida converts (S)-3-ethoxy-4-hydroxymandelic acid into ethyl vanillin, but does not degrade ethyl vanillin, overview
-
-
-
additional information
?
-
-
synthesis of (S)-2-hydroxy-1-phenylpropanone by the immobilized recombinant enzyme is effective in cross acyloin reaction of benzaldehyde with acetaldehyde. The immobilized and free enzymes shows similar substrate specificity and activity
-
-
-
additional information
?
-
-
the enzyme also performs the cross acyloin reaction of benzaldehyde with acetaldehyde to give (S)-2-hydroxy-1-phenylpropanone
-
-
-
additional information
?
-
-
the enzyme has the ability to carry out stereospecific carbon-carbon bond formation, with products including various 2-hydroxy ketones and benzoin derivatives, the enzyme also catalyzes the conversion of benzaldehyde and acetaldehyde into R-benzoin and, predominantly, S-2-hydroxypropiophenone, via stereospecific carboligation
-
-
-
additional information
?
-
when 3-ethoxy-4-hydroxybenzoylformate is provided at with Pseudomonas putida, it is completely transformed to ethyl vanillic acid after 2 days, but only trace amounts of ethyl vanillin are detected. Similar results are obtained when ethyl vanillin replaces 3-ethoxy-4-hydroxybenzoylformate as substrate, thus aldehyde dehydrogenation activity in Pseudomonas putida is greater than its synthesis activity. The recombinant Escherichia coli strain expressing the enzyme from Pseudomonas putida converts (S)-3-ethoxy-4-hydroxymandelic acid into ethyl vanillin, but does not degrade ethyl vanillin, overview
-
-
-
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
3-ethoxy-4-hydroxybenzoylformate
ethyl vanillin + CO2
P20906
-
-
-
?
Benzoylformate
?
-
enzyme is involved in the inducible pathway of mandelate
-
-
-
Benzoylformate
?
-
enzyme is involved in the metabolism of mandelic acid
-
-
-
Benzoylformate
?
-
enzyme is involved in the metabolism of mandelic acid
-
-
-
Benzoylformate
?
-
enzyme is involved in the inducible pathway of mandelate
-
-
-
Benzoylformate
?
-
enzyme is involved in the metabolism of mandelic acid
-
-
-
Benzoylformate
?
-
enzyme is involved in the metabolic pathway of phenylglycine
-
-
-
Benzoylformate
?
-
enzyme is involved in the metabolic pathway of phenylglycine
-
-
-
Benzoylformate
?
-
the enzyme is involved in the pathway for catabolism of D(+)-mandelate and L-(-)-mandelate
-
-
-
additional information
?
-
-
the enzyme performs enantioselective synthesis of (S)-2-hydroxyketones. Its stereoselectivity is highly dependent on the structure of the substrate aldehydes
-
-
-
additional information
?
-
P20906
when 3-ethoxy-4-hydroxybenzoylformate is provided at with Pseudomonas putida, it is completely transformed to ethyl vanillic acid after 2 days, but only trace amounts of ethyl vanillin are detected. Similar results are obtained when ethyl vanillin replaces 3-ethoxy-4-hydroxybenzoylformate as substrate, thus aldehyde dehydrogenation activity in Pseudomonas putida is greater than its synthesis activity. The recombinant Escherichia coli strain expressing the enzyme from Pseudomonas putida converts (S)-3-ethoxy-4-hydroxymandelic acid into ethyl vanillin, but does not degrade ethyl vanillin, overview
-
-
-
additional information
?
-
P20906
when 3-ethoxy-4-hydroxybenzoylformate is provided at with Pseudomonas putida, it is completely transformed to ethyl vanillic acid after 2 days, but only trace amounts of ethyl vanillin are detected. Similar results are obtained when ethyl vanillin replaces 3-ethoxy-4-hydroxybenzoylformate as substrate, thus aldehyde dehydrogenation activity in Pseudomonas putida is greater than its synthesis activity. The recombinant Escherichia coli strain expressing the enzyme from Pseudomonas putida converts (S)-3-ethoxy-4-hydroxymandelic acid into ethyl vanillin, but does not degrade ethyl vanillin, overview
-
-
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
-
the addition of thiamine thiazolone diphosphate has no effect on the activity of BFD in the presence of thiamine diphosphate
-
thiamine diphosphate
-
the 4'-aminopyrimidine ring of the thiamine diphosphate coenzyme participates in catalysis, likely as an intramolecular general acid-base catalyst via the unusual 1',4'-iminopyrimidine tautomer
thiamine diphosphate
-
the enzyme avoids formation of a benzoyl anion by addition of enzyme-bound thiamine diphosphate to the carbonyl group of benzoylformate, producing enzyme-bound 2-(2-mandelyl)thiamine diphosphate
thiamine diphosphate
-
-
thiamine diphosphate
-
dependent on, structural binding environment, overview. The V-conformation is assisted by the presence of a fulcrum residue, Leu403, located directly below the cofactor, the V-conformation results in a N4'-C2 distance of 3.1 A
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
Mn2+
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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
benzoylphosphonate
-
the reaction of methyl benzoylphosphonate with BFDC results in the formation of a stable analogue (C2alpha-phosphonomandelyl thiamine diphosphate) of the covalent thiamine diphosphate-substrate adduct C2alpha-mandelyl thiamine diphosphate
[p-(Bromomethyl)benzoyl]formate
-
due to covalent modification of the cofactor
[p-(Bromomethyl)benzoyl]formate
-
modification of thiamine diphosphate by halide elimination and tautomerization
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
KM-value of wild-type enzyme for 2-oxo-4-methylhexanoic acid (pH 6.0, 30°C) is above 20 mM
-
0.42
2-ketobutanoate
-
mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
1.3
2-ketobutanoate
-
mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
4.3
2-ketobutanoate
-
mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
7.5
2-ketobutanoate
-
wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.15
2-ketohexanoate
-
mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.28
2-ketohexanoate
-
mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
1.3
2-ketohexanoate
-
mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
4.1
2-ketohexanoate
-
wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.1
2-ketopentanoate
-
mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.28
2-ketopentanoate
-
mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
1.1
2-ketopentanoate
-
mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
6
2-ketopentanoate
-
wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
515.9
acetaldehyde
-
immobilized mutant enzyme A460I/F464I, in 50 mM phosphate buffer at pH 7.5 and 20°C
775
acetaldehyde
-
free mutant enzyme A460I/F464I, in 50 mM phosphate buffer at pH 7.5 and 20°C
19.4
benzaldehyde
-
free mutant enzyme A460I/F464I, in 50 mM phosphate buffer at pH 7.5 and 20°C
51.9
benzaldehyde
-
immobilized mutant enzyme A460I/F464I, in 50 mM phosphate buffer at pH 7.5 and 20°C
0.06
benzoylformate
-
mutant enzyme L461V, in potassium phosphate buffer (pH 6.5, 50 mM)
0.23
benzoylformate
-
in 50 mM potassium phosphate buffer, pH 6.5, at 30°C
0.25
benzoylformate
-
mutant enzyme L461A, in potassium phosphate buffer (pH 6.5, 50 mM)
0.27
benzoylformate
-
wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.29
benzoylformate
-
mutant enzyme H281Q, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C; mutant enzyme H281W, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.37
benzoylformate
-
wild type enzyme, in potassium phosphate buffer (pH 6.5, 50 mM)
0.38
benzoylformate
-
mutant enzyme H70L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.49
benzoylformate
-
mutant enzyme H281Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.51
benzoylformate
-
mutant enzyme P24A, in potassium phosphate buffer (pH 6.5, 50 mM)
0.54
benzoylformate
-
wild type enzyme, in 50 mM potassium phosphate buffer pH 7.0, 0.5 mM thiamine diphosphate, 2.5 mM MgSO4, at 30°C
0.54
benzoylformate
-
mutant enzyme H281F, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.58
benzoylformate
-
mutant enzyme L476Q, in 50 mM potassium phosphate buffer pH 7.0, 0.5 mM thiamine diphosphate, 2.5 mM MgSO4, at 30°C
0.59
benzoylformate
-
mutant enzyme L476G, in 50 mM potassium phosphate buffer pH 7.0, 0.5 mM thiamine diphosphate, 2.5 mM MgSO4, at 30°C
0.6
benzoylformate
-
mutant enzyme S181T/L476P, in 50 mM potassium phosphate buffer pH 7.0, 0.5 mM thiamine diphosphate, 2.5 mM MgSO4, at 30°C
0.8
benzoylformate
-
mutant enzyme S26M, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.85
benzoylformate
-
mutant enzyme H70F, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C; mutant enzyme S26T, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.89
benzoylformate
-
mutant enzyme H70T, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.9
benzoylformate
-
mutant enzyme H281T, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.94
benzoylformate
-
mutant enzyme H70S, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
1
benzoylformate
-
mutant enzyme L476H, in 50 mM potassium phosphate buffer pH 7.0, 0.5 mM thiamine diphosphate, 2.5 mM MgSO4, at 30°C
1.02
benzoylformate
-
mutant enzyme L476T, in 50 mM potassium phosphate buffer pH 7.0, 0.5 mM thiamine diphosphate, 2.5 mM MgSO4, at 30°C
1.03
benzoylformate
-
mutant enzyme L476P, in 50 mM potassium phosphate buffer pH 7.0, 0.5 mM thiamine diphosphate, 2.5 mM MgSO4, at 30°C
1.06
benzoylformate
-
mutant enzyme L476Q/S525G, in 50 mM potassium phosphate buffer pH 7.0, 0.5 mM thiamine diphosphate, 2.5 mM MgSO4, at 30°C
1.08
benzoylformate
-
mutant enzyme L476M, in 50 mM potassium phosphate buffer pH 7.0, 0.5 mM thiamine diphosphate, 2.5 mM MgSO4, at 30°C
1.14
benzoylformate
-
mutant enzyme L476S, in 50 mM potassium phosphate buffer pH 7.0, 0.5 mM thiamine diphosphate, 2.5 mM MgSO4, at 30°C
1.17
benzoylformate
-
mutant enzyme L476A, in 50 mM potassium phosphate buffer pH 7.0, 0.5 mM thiamine diphosphate, 2.5 mM MgSO4, at 30°C
1.2
benzoylformate
-
mutant enzyme H281A, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C; mutant enzyme S26L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
1.23
benzoylformate
-
mutant enzyme S181T, in 50 mM potassium phosphate buffer pH 7.0, 0.5 mM thiamine diphosphate, 2.5 mM MgSO4, at 30°C
1.26
benzoylformate
-
mutant enzyme L476C, in 50 mM potassium phosphate buffer pH 7.0, 0.5 mM thiamine diphosphate, 2.5 mM MgSO4, at 30°C
1.4
benzoylformate
-
mutant enzyme L461G, in potassium phosphate buffer (pH 6.5, 50 mM)
1.46
benzoylformate
-
mutant enzyme L476K, in 50 mM potassium phosphate buffer pH 7.0, 0.5 mM thiamine diphosphate, 2.5 mM MgSO4, at 30°C
1.5
benzoylformate
-
mutant enzyme H70A, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
1.54
benzoylformate
-
mutant enzyme A460G, in potassium phosphate buffer (pH 6.5, 50 mM)
2.9
benzoylformate
-
mutant enzyme H281N in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
7.8
benzoylformate
-
mutant enzyme S26A, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.04
benzylformate
-
mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.27
benzylformate
-
wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.45
benzylformate
-
mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
1.5
benzylformate
-
mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.27
phenylglyoxylate
-
pH and temperature not specified in the publication, wild-type enzyme
0.38
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H70L
0.49
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H281Y
0.54
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H281F
0.8
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant S26M
0.85
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H70F
1.2
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H281A; pH and temperature not specified in the publication, mutant S26L
1.5
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H70A
7.8
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant S26A
2
pyruvate
-
mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
4.8
pyruvate
-
mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
21
pyruvate
-
mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
4
2-ketobutanoate
-
wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
5.8
2-ketobutanoate
-
mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
6
2-ketobutanoate
-
mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
7.5
2-ketobutanoate
-
mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
1.1
2-ketohexanoate
-
mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
1.2
2-ketohexanoate
-
mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
4.4
2-ketohexanoate
-
mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
5.3
2-ketohexanoate
-
wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.81
2-ketopentanoate
-
mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
3.3
2-ketopentanoate
-
mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
6.2
2-ketopentanoate
-
mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
11
2-ketopentanoate
-
wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.46
benzoylformate
-
mutant enzyme H70A, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
2
benzoylformate
-
mutant enzyme H281T, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
2.1
benzoylformate
-
mutant enzyme H281A, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
4.5
benzoylformate
-
mutant enzyme H70F, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
5.6
benzoylformate
-
mutant enzyme H70S, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
6.9
benzoylformate
-
mutant enzyme H281Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
8.6
benzoylformate
-
mutant enzyme H281Q, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
14
benzoylformate
-
mutant enzyme H70L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
15
benzoylformate
-
mutant enzyme S26A, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
17
benzoylformate
-
mutant enzyme H281W, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
19
benzoylformate
-
mutant enzyme H281N in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
26
benzoylformate
-
mutant enzyme S26M, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
42
benzoylformate
-
mutant enzyme H70T, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
65
benzoylformate
-
mutant enzyme H281F, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
94
benzoylformate
-
mutant enzyme S26T, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
132
benzoylformate
-
mutant enzyme S26L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
320
benzoylformate
-
wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
1.7
benzylformate
-
mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
25
benzylformate
-
mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
110
benzylformate
-
mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
320
benzylformate
-
wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.46
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H70A
2.1
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H281A
4.5
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H70F
6.9
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H281Y
14
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H70L
15
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant S26A
26
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant S26M
65
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H281F
132
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant S26L
320
phenylglyoxylate
-
pH and temperature not specified in the publication, wild-type enzyme
1.4
pyruvate
-
mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
2
pyruvate
-
mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
3.6
pyruvate
-
mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.5
2-ketobutanoate
-
wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
1.7
2-ketobutanoate
-
mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
4.6
2-ketobutanoate
-
mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
14
2-ketobutanoate
-
mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.86
2-ketohexanoate
-
mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
1.3
2-ketohexanoate
-
wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
8
2-ketohexanoate
-
mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
16
2-ketohexanoate
-
mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
1.8
2-ketopentanoate
-
wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
3
2-ketopentanoate
-
mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
8.1
2-ketopentanoate
-
mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
22
2-ketopentanoate
-
mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
16
benzylformate
-
mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
43
benzylformate
-
mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
244
benzylformate
-
mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
1180
benzylformate
-
wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.3
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H70A
1.7
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H281A
1.9
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant S26A
5.3
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H70F
14
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H281Y
32
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant S26M
36
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H70L
110
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant S26L
120
phenylglyoxylate
-
pH and temperature not specified in the publication, mutant H281F
1180
phenylglyoxylate
-
pH and temperature not specified in the publication, wild-type enzyme
0.012
pyruvate
-
wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.065
pyruvate
-
mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
0.42
pyruvate
-
mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
1.8
pyruvate
-
mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
20
2-Oxopentanoic acid
-
pH 6.0, 30°C, wild-type enzyme and mutant enzyme A460I
0.36
benzoylphosphonate
-
prolonged incubation of BFD with benzoylphosphonate results in permanent inactivation of the enzyme
40
(R)-mandelate
-
pH 6.5, mutant enzyme H281A; pH 6.5, mutant enzyme H70A
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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 30°C 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.
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6.2
8.5
7
-
carboligation yielding (S)-2-hydroxy-1-phenyl-propanone with benzoylformate and acetaldehyde as substrate
8.5
-
carboligation yielding (S)-2-hydroxy-1-phenyl-propanone with benzaldehyde and acetaldehyde as substrates
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
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
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30
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TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
20 - 70
-
At temperatures above 70°C and lower than 20°C the activity decreases below 10 U/g carrier.
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
no activity with benzoate or glucose as growth substrate
-
no activity with benzoate or glucose as growth substrate
-
no activity with benzoate or glucose as growth substrate
-
no activity with benzoate or glucose as growth substrate
-
high activity, no activity with succinate as growth substrate
-
high activity, no activity with succinate as growth substrate
-
-
high activity, no activity with succinate as growth substrate
-
high activity, no activity with succinate as growth substrate
-
-
no activity with glucose as growth substrate
-
no actiity with succinate as growth substrate
-
no actiity with succinate as growth substrate
-
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PDB
SCOP
CATH
ORGANISM
UNIPROT
Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)
Polynucleobacter asymbioticus (strain DSM 18221 / CIP 109841 / QLW-P1DMWA-1)
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homotetramer
tetramer
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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
-
sitting drop vapor diffusion method, using 25% (w/v) polyethylene glycol 2000 and 0.2 M MgCl2, at pH 6.
-
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
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
-
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ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
DMSO
-
retains its full activity after 10 days of incubation in DMSO (20% v/v) at 25°C
Ethanol
-
retains its full activity after 10 days of incubation in aqueous ethanol (1.5 M) at 25°C
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
20°C, pH 7, in presence of 0.5 mM thiamin diphosphate and 0.5 mM Mg2+, half-life is 36 d
-
20°C, pH 7, in presence of 0.5 mM thiamin diphosphate, 100 mM acetaldehyde and 0.5 mM Mg2+, half-life is 1.8 d
-
20°C, pH 7, in presence of 0.5 mM thiamin diphosphate, 200 mM acetaldehyde and 0.5 mM Mg2+, half-life is 1.5 d
-
20°C, pH 7, in presence of 0.5 mM thiamin diphosphate, 25 mM benzaldehyde and 0.5 mM Mg2+, half-life is 5.3 d
-
4°C, pH 7, in presence of 0.5 mM thiamin diphosphate and 0.5 mM Mg2+, half-life is 139 d
-
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Phenyl Sepharose FF column chromatography and Q Sepharose FF column chromatography
-
Q-Sepharose column chromatography and 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
-
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
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
-
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A460G
-
mutant exhibits a higher Km value compared to the wild type enzyme
A460I
A460I/F464I
A460Y
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
H281A
H281F
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
H281Y
H70A
H70F
H70L
H70S
-
mutant exhibits a 197fold decrease in kcat/Km compared to the wild type enzyme
H70T
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
L461A
L461G
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
L476H
L476M
-
has a 4.23fold higher carboligase activity than the wild type enzyme
L476P
L476Q
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
S181T
S26A
S26L
S26M
S26T
-
mutant exhibits no significant loss of activity compared to the wild type enzyme (3fold decrease in kcat value)
T377L
T377L/A460Y
additional information
A460I
-
activity with pyruvate is higher than wild-type activity, activity with 2-oxobutanoic acid is 2.3fold higher than wild-type activity, activity with 2-oxopentanoic acid is 1.1fold lower than wild-type activity, activity with 2-oxohexanoic acid is 3.3fold higher than wild-type activity, activity with 3-methyl-2-oxobutanoic acid is 1.3fold higher than wild-type activity, activity with 3-methyl-2-oxopentanoic acid is 1.1fold lower than wild-type activity, activity with 2-oxo-4-methylpentanoic acid is 1.3fold 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 2.2fold higher than wild-type activity, activity with benzoylformate is 22.5fold 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 is 1.8fold higher than wild-type activity, activity with 2-oxo-5-phenylpentanoic acid is identical to wild-type activity. The ratio of turnover number to KM-value for 2-oxo-4-methylpentanoic acid is 10fold higher than the wild-type value, the ratio of turnover number to KM-value for 2-oxo-4-methylhexanoic acid is 1.13fold higher than the wild-type value, the ratio of turnover number to KM-value for benzoylformate is 5.4fold lower than the wild-type value, the ratio of turnover number to KM-value for 2-oxohexanoic acid is 14.1fold higher than the wild-type value, the ratio of turnover number to KM-value for 2-oxopentanoic acid is 4.6fold higher 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
A460I
-
for this variant the highest amounts of (S)-2-hydroxypropiophenone-product can be detected at low benzaldehyde concentrations and slightly acidic conditions at pH 6.5
A460I
-
the mutant exhibits an increased (R)-2-hydroxypropiophenone selectivity
A460I
-
site-directed mutagenesis, the mutant enzymes is an excellent 2-ketohexanoate decarboxylase
A460I/F464I
-
activity with pyruvate is higher than wild-type activity, activity with 2-oxobutanoic acid is 1.8fold lower than wild-type activity, activity with 2-oxopentanoic acid is 9.2fold lower than wild-type value, activity with 2-oxohexanoic acid is 3.3fold lower than wild-type activity, activity with 3-methyl-2-oxobutanoic acid is 1.5fold lower than wild-type activity, activity with 3-methyl-2-oxopentanoic acid is 1.2fold lower than wild-type activity, activity with 2-oxo-4-methylpentanoic acid is 1.5fold lower than wild-type activity, activity with 4,4-dimethyl-2-oxopentanoic acid is 10fold lower than wild-type activity, activity with 2-oxo-4-methylhexanoic acid is 1.1fold lower than wild-type activity, activity with benzoylformate is 64.3fold lower than wild-type activity, no activity with 2-oxo-3-phenylpropanoic acid, activity with 2-oxo-4-phenylbutanoic acid is 1.3fold lower than wild-type activity, activity with 2-oxo-5-phenylpentanoic acid is fold higher than wild-type activity. the ratio of turnover number to KM-value for benzoylformate is 88.7fold lower than the wild-type value
A460I/F464I
-
for this variant the highest amounts of (S)-2-hydroxypropiophenone-product can be detected at low benzaldehyde concentrations and slightly acidic conditions at pH 6.5
A460I/F464I
-
the mutant exhibits an increased (R)-2-hydroxypropiophenone selectivity
A460Y
-
the mutant shows significantly improved catalytic activity toward pyruvate
A460Y
-
site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme
F464I
-
for this variant the highest amounts of (S)-2-hydroxypropiophenone-product can be detected at low benzaldehyde concentrations and slightly acidic conditions at pH 6.5
F464I
-
the mutant exhibits an increased (R)-2-hydroxypropiophenone selectivity
H281A
-
4.5fold increase in Km-value for benzoylformate, 3443fold decrease in turnover number for benzoylformate, 171fold increase in Ki-value for (R)-mandelate compared to wild-type enzyme
H281A
-
mutant with 152fold decreased kcat value compared to the wild type enzyme
H281A
-
site-directed mutagenesis, the mutant shows about 600fold reduced catalytic efficiency compared to the wild-type enzyme
H281F
-
mutant with 4.9fold decrease in kcat value compared to the wild type enzyme
H281F
-
site-saturation mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
H281Y
-
mutant with 46fold decrease in kcat value compared to the wild type enzyme
H281Y
-
site-saturation mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
H70A
-
5.2fold increase in Km-value for benzoylformate, 3443fold decrease in turnover number for benzoylformate, 40fold increase in Ki-value for (R)-mandelate compared to wild-type enzyme
H70A
-
mutant exhibits 4000fold decreased catalytic activity compared to the wild type enzyme
H70A
-
site-directed mutagenesis, the mutant shows about 4000fold reduced catalytic efficiency compared to the wild-type enzyme
H70F
-
mutant exhibits a 236fold decrease in kcat/Km compared to the wild type enzyme
H70F
-
site-saturation mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
H70L
-
mutant exhibits a 33fold decrease in kcat/Km compared to the wild type enzyme
H70L
-
site-saturation mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
H70T
-
mutant exhibits a 25fold decrease in kcat/Km compared to the wild type enzyme
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)
L461A
-
the mutant has decreased activity compared to the wild type enzyme, the enantioselectivity of the (S)-2-hydroxypropiophenone formation is increased by 6%. In the case of propionaldehyde, the enzyme activity increases considerably from 6% with wild type BFD to 21.5%. With butyraldehyde the already low enzyme activity does not change
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)
L461G
-
the mutant has decreased activity compared to the wild type enzyme, the enantioselectivity of the (S)-2-hydroxypropiophenone formation is increased by 6%. In the case of propionaldehyde, the enzyme activity increases considerably from 6% with wild type BFD to 23%. With butyraldehyde the already low enzyme activity does not change
L476H
-
has a 3.19fold higher carboligase activity than the wild type enzyme
L476P
-
the mutant variant shows improved carboligase activity in organic solvents compared to the wild-type enzyme
L476Q
-
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. Vmax and Km-value for decraboxylation of benzoylformate are not effected
L476Q
-
has a 5.06fold higher carboligase activity than the wild type enzyme
L476Q
-
the mutant variant shows improved carboligase activity in organic solvents compared to the wild-type enzyme
S181T
-
has a 1.75fold higher carboligase activity than the wild type enzyme
S26A
-
23.1fold increase in Km-value for benzoylformate, 54fold decrease in turnover number for benzoylformate, 100fold increase in Ki-value for (R)-mandelate compared to wild-type enzyme
S26A
-
mutant with 21fold decrease in kcat value compared to the wild type enzyme
S26A
-
site-directed mutagenesis, the mutant shows about 600fold reduced catalytic efficiency compared to the wild-type enzyme, the S26A variant showed a 30fold increase in Km for benzoylformate and a 100fold increase in Ki value for R-mandelate
S26L
-
mutant exhibits no significant loss of activity compared to the wild type enzyme (2fold decrease in kcat value)
S26L
-
site-saturation mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
S26M
-
mutant exhibits no significant loss of activity compared to the wild type enzyme (12fold decrease in kcat value)
S26M
-
site-saturation mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
T377L
-
the mutant shows significantly improved catalytic activity toward pyruvate
T377L
-
site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme
T377L/A460Y
-
the double mutant shows significantly improved catalytic activity toward pyruvate (11000fold improvement in the ratio between pyruvate and benzoylformate utilization)
T377L/A460Y
-
site-directed mutagenesis, the mutant shows altered substrate specificity and kinetics compared to the wild-type enzyme
additional information
biotransformation using recycled resting Escherichia coli cells expressing the enzyme, the product yield is above 42% after four repeated cycles
additional information
-
immobilization of the purified recombinant His-tagged enzyme on magnetic nanoparticles, microspheres, nanospheres and ferrofluids, for enantioselective synthesis of (S)-2-hydroxyketones. Its stereoselectivity is highly dependent on the structure of the substrate aldehydes. Adsorption capacity of the Cu+2 charged magnetic solid support for His-tagged BFD is determined by standard BSA assay as 43.6 mg enzyme/g magnetic solid support
additional information
-
site-saturation mutagenesis is used to evolve stereoselective enzymes as catalysts for synthetic organic chemistry
additional information
-
covalent immobilization and stabilization of benzoylformate decarboxylase from Pseudomonas putida on magnetic nanoparticles by a three-step immobilization/stabilization procedure at pH 8.0, 20°C, method development and evaluation, overview. The maximum enzyme loading is found to be 5.2 mg/g support in 3.5 mg/ml initial enzyme concentration at ionic strength of 1.25 M MgSO4. The covalently-bound enzyme is characterized in terms of its activity and stability for the formation of (S)-2-hydroxypropiophenone. The immobilized biocatalyst retains 95% of its original activity after five reaction cycles
additional information
-
biotransformation using recycled resting Escherichia coli cells expressing the enzyme, the product yield is above 42% after four repeated cycles
-
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
biotechnology
-
Production of immobilized enzyme on Sepabeads EC-EA by C-C coupling
synthesis
synthesis
-
synthesis of enantiopure unsymmetrical and symmetrical 2-hydroxy ketones
synthesis
-
enzyme-catalyzed synthesis of enantiomerically pure (R)-benzoin derivatives in good yield
synthesis
the enzyme recombinantly expressed in Escherichia coli is useful for production of ethyl vanillin, i.e. 3-ethoxy-4-hydroxybenzaldehyde, which is a good substitute for vanillin in foods and perfumes since it is 3.5times stronger in flavor and keeps better condition in storage and transport
synthesis
-
purified recombinant His-tagged enzyme immobilized on magnetic nanoparticles, microspheres, nanospheres and ferrofluids, is used for enantioselective synthesis of (S)-2-hydroxyketones. Its stereoselectivity is highly dependent on the structure of the substrate aldehydes. The heterogeneous biocatalyst is offering the ease of immobilization along with the separation steps of metal affinity ligand (Cu2+-iminodiacetic acid) coated magnetic nanoparticles. Reusability of immobilized enzyme for carboligation reactions
synthesis
-
the enzyme recombinantly expressed in Escherichia coli is useful for production of ethyl vanillin, i.e. 3-ethoxy-4-hydroxybenzaldehyde, which is a good substitute for vanillin in foods and perfumes since it is 3.5times stronger in flavor and keeps better condition in storage and transport
-
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LINKS TO OTHER DATABASES (specific for EC-Number 4.1.1.7)
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