4.1.1.7: benzoylformate decarboxylase
This is an abbreviated version!
For detailed information about benzoylformate decarboxylase, go to the full flat file.
Word Map on EC 4.1.1.7
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4.1.1.7
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thiamin
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putida
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thdp-dependent
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diphosphate-dependent
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carboligation
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carboligase
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acyloin
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r-benzoin
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synthesis
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s-mandelate
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2-keto
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biotechnology
- 4.1.1.7
- thiamin
- putida
-
thdp-dependent
-
diphosphate-dependent
-
carboligation
-
carboligase
- acyloin
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r-benzoin
- synthesis
- s-mandelate
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2-keto
- biotechnology
Reaction
Synonyms
benzoylformate decarboxylase, BFD, BfdB, BFDC, BfdM, Decarboxylase, benzoylformate, MdlC, Phenylglyoxylate decarboxylase
ECTree
4.1.1.7 benzoylformate decarboxylaseAdvanced search results
results (
results (Engineering
Engineering on EC 4.1.1.7 - benzoylformate decarboxylase
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
I466A
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
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kcat value of the L403E variant is only 18fold lower than that of wild-type BFDC
L403F
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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
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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
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has a 4.11fold higher carboligase activity than the wild type enzyme
L476C
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has a 4.31fold higher carboligase activity than the wild type enzyme
L476G
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has a 4.27fold higher carboligase activity than the wild type enzyme
L476H
L476M
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has a 4.23fold higher carboligase activity than the wild type enzyme
L476P
L476Q
L476S
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has a 3.5fold higher carboligase activity than the wild type enzyme
L476T
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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
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
I466A
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the mutant shows increased catalytic efficiency for benzoylformate compared to the wild type enzyme
I466A
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the mutant shows increased catalytic efficiency for benzoylformate compared to the wild type enzyme
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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
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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
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the mutant exhibits an increased (R)-2-hydroxypropiophenone selectivity
A460I
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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
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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
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the mutant exhibits an increased (R)-2-hydroxypropiophenone selectivity
A460Y
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site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme
A460Y
the mutant shows significantly improved catalytic activity toward pyruvate
F464I
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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
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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
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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
the mutation leads to a 5fold decrease in the enzyme activity
H281F
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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
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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
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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
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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
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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
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has a 3.19fold higher carboligase activity than the wild type enzyme
L476P
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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
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has a 5.06fold higher carboligase activity than the wild type enzyme
L476Q
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the mutant variant shows improved carboligase activity in organic solvents compared to the wild-type enzyme
S181T
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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
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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
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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
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site-saturation mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
T377L
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site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme
T377L
the mutant shows significantly improved catalytic activity toward pyruvate
T377L/A460Y
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site-directed mutagenesis, the mutant shows altered substrate specificity and kinetics 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
the mutant exhibits a greater than 10000fold increase in pyruvate/benzoylformate substrate utilization ratio compared to that of 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
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biotransformation using recycled resting Escherichia coli cells expressing the enzyme, the product yield is above 42% after four repeated cycles
additional information
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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
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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
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site-saturation mutagenesis is used to evolve stereoselective enzymes as catalysts for synthetic organic chemistry
additional information
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biotransformation using recycled resting Escherichia coli cells expressing the enzyme, the product yield is above 42% after four repeated cycles
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