Information on EC 4.1.2.47 - (S)-hydroxynitrile lyase

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The expected taxonomic range for this enzyme is: Malpighiales

EC NUMBER
COMMENTARY hide
4.1.2.47
-
RECOMMENDED NAME
GeneOntology No.
(S)-hydroxynitrile lyase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
an aliphatic (S)-hydroxynitrile = cyanide + an aliphatic aldehyde or ketone
show the reaction diagram
an aromatic (S)-hydroxynitrile = cyanide + an aromatic aldehyde
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
condensation
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cyanohydrin formation
Henry reaction
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
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Cyanoamino acid metabolism
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SYSTEMATIC NAME
IUBMB Comments
(S)-cyanohydrin lyase (cyanide forming)
Hydroxynitrile lyases catalyses the the cleavage of hydroxynitriles into cyanide and the corresponding aldehyde or ketone. In nature the liberation of cyanide serves as a defense mechanism against herbivores and microbial attack in plants. In vitro the enzymes from Manihot esculenta and Hevea brasiliensis accept a broad range of aliphatic and aromatic carbonyl compounds as substrates and catalyse the formation of (S)-hydroxynitriles [1,10].
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
flax
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Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(1S)-1-furan-2-yl-2-nitroethanol
furan-2-carbaldehyde + CH3NO2
show the reaction diagram
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enantiomeric excess: ~90%, yield: 60-70%
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r
(1S)-2-nitro-1-(4-nitrophenyl)ethanol
4-nitrobenzaldehyde + CH3NO2
show the reaction diagram
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enantiomeric excess: ~90%, yield: 60-70%
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r
(1S)-2-nitro-1-phenylethanol
benzaldehyde + CH3NO2
show the reaction diagram
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enantiomeric excess: ~90%, yield: 60-70%
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r
(1S,2R)-2-nitro-1-phenyl-propanol
benzaldehyde + C2H5NO2
show the reaction diagram
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4 diastereomers, enantiomeric excess: 95%, yield: 67%
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r
(2S)-1-nitrooctan-2-ol
heptanal + CH3NO2
show the reaction diagram
-
enantiomeric excess: ~90%, yield: 60-70%
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r
(2S)-2,3-dimethyl-2-hydroxybutyronitrile
?
show the reaction diagram
-
binding mode of the chiral substrates is identical to that observed for the biological substrate 2-hydroxy-2-methylpropanenitrile (i.e. acetone cyanohydrin). Three-point binding mode of the substrates: hydrophobic pocket, hydrogen bonds between the hydroxyl group and Ser80 and Thr11, electrostatic interaction of the cyano group with Lys236
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?
(2S)-2-hydroxy-2-methylbutanenitrile
cyanide + butan-2-one
show the reaction diagram
(2S)-2-hydroxy-2-methylpentanenitrile
cyanide + pentan-2-one
show the reaction diagram
-
-
-
?
(2S)-hydroxy(3-phenoxyphenyl)ethanenitrile
cyanide + 3-phenoxybenzaldehyde
show the reaction diagram
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r
(R)-2-(2-furyl)-2-hydroxyacetonitrile
furan-2-carbaldehyde + HCN
show the reaction diagram
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enantiomeric excess: > 99%, yield: 90%
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r
(S)-2-nitro-1-phenylethanol
benzaldehyde + nitromethane
show the reaction diagram
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r
(S)-2-nitro-1-phenylethanol
nitromethane + benzaldehyde + (R)-2-nitro-1-phenylethanol
show the reaction diagram
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besides the native cyanohydrins reaction, the enzyme also catalyzes the asymmetric reversible Henry reaction yielding (S)-beta-nitroalcohols with high enantiomeric excess. The catalyst productivity achieved during the resolution is 10times higher than that in the HNL-catalyzed synthesis of (S)-2-nitro-1-phenylethanol
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r
(S)-3-phenoxybenzaldehyde cyanohydrin
m-phenoxybenzaldyhyde + HCN
show the reaction diagram
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enantiomeric excess: > 98.5%, yield: 95.5%, used for insecticide synthesis
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r
(S)-mandelonitrile
benzaldehyde + HCN
show the reaction diagram
(S)-Mandelonitrile
Cyanide + benzaldehyde
show the reaction diagram
(S)-mandelonitrile
cyanide + benzaldehyde + HCN
show the reaction diagram
-
the enzyme is a member of the alpha/beta hydrolase fold protein family, containing a catalytic triad with C-C cleaving and ligating activity
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r
(S)-mandelonitrile
HCN + benzaldehyde
show the reaction diagram
2-furaldehyde cyanohydrin
2-furaldehyde + HCN
show the reaction diagram
2-Furylaldehyde + cyanide
Furan-3-yl-hydroxyacetonitrile
show the reaction diagram
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2-hydroxy-2-methylpropanenitrile
acetone + HCN
show the reaction diagram
2-hydroxy-2-methylpropanenitrile
cyanide + acetone
show the reaction diagram
2-hydroxy-2-methylpropanenitrile
HCN + acetone
show the reaction diagram
2-hydroxyisobutyronitrile
HCN + acetone
show the reaction diagram
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-
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?
2-Methyl-2-hydroxybutyronitrile
Butanone + cyanide
show the reaction diagram
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2-nitro-1-phenylethanol
?
show the reaction diagram
2-Pentanone + cyanide
3-Hydroxyhexanonitrile
show the reaction diagram
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2-Thienylaldehyde + cyanide
Hydroxythiophen-3-yl-acetonitrile
show the reaction diagram
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3,3-dimethyl-2-butanone + acetone cyanohydrin
(S)-2-hydroxy-2-methyl-3,3-dimethyl-butyronitrile
show the reaction diagram
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transcyanation
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?
3-[(1S)-1-hydroxy-2-nitroethyl]phenol
3-hydroxybenzaldehyde + CH3NO2
show the reaction diagram
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enantiomeric excess: ~90%, yield: 60-70%
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r
4-Methoxybenzaldehyde + cyanide
4-Methoxymandelonitrile
show the reaction diagram
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Acetone cyanhydrin
Cyanide + acetone
show the reaction diagram
acetone cyanhydrin
HCN + acetone
show the reaction diagram
acetone cyanohydrin
cyanide + acetone
show the reaction diagram
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?
acetone cyanohydrin
hydrocyanic acid + acetone
show the reaction diagram
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?
Acetophenone + cyanide
3-Hydroxy-3-phenylpropionitrile
show the reaction diagram
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acetyltrimethylsilane + acetone cyanohydrin
(S)-2-trimethylsilyl-2-hydroxyl-propionitrile + acetone
show the reaction diagram
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transcyanation
-
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?
Benzaldehyde + cyanide
(S)-Mandelonitrile
show the reaction diagram
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cyanide + (2E)-3-(4-hydroxyphenyl)prop-2-enal
(2S,3E)-2-hydroxy-4-(4-hydroxyphenyl)but-3-enenitrile
show the reaction diagram
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wild-type enzyme: 95% enantiomeric excess, 80% conversion rate
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?
cyanide + (2E)-but-2-enal
(3E)-2-hydroxypent-3-enenitrile
show the reaction diagram
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86% enantiomeric excess with crude enzyme preparation
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?
cyanide + (2E)-but-2-enal
(3S,3E)-2-hydroxypent-3-enenitrile
show the reaction diagram
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92% enantiomeric excess
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?
cyanide + (2E)-hex-2-enal
(2S,3E)-2-hydroxyhept-3-enenitrile
show the reaction diagram
cyanide + (2Z)-hex-2-enal
(2S,3Z)-2-hydroxyhept-3-enenitrile
show the reaction diagram
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80% enantiomeric excess with crude enzyme preparation
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?
cyanide + (4-hydroxyphenyl)acetaldehyde
(2S)-2-hydroxy-3-(4-hydroxyphenyl)propanenitrile
show the reaction diagram
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wild-type enzyme: 96% enantiomeric excess, 88% conversion rate
-
?
cyanide + (4R)-2,2-dimethyl-1,3-dioxolane-4-carbaldehyde
(2S)-[(4R)-2,2-dimethyl-1,3-dioxolan-4-yl](hydroxy)ethanenitrile + (2R)-[(4R)-2,2-dimethyl-1,3-dioxolan-4-yl](hydroxy)ethanenitrile
show the reaction diagram
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the natural substrate benzaldehyde is stereoselectively converted to (R)-mandelonitrile. The non-natural substrate (4R)-2,2-dimethyl-1,3-dioxolane-4-carbaldehyde is converted to 47.1% (2S)-[(4R)-2,2-dimethyl-1,3-dioxolan-4-yl](hydroxy)ethanenitrile and 52.9% (2R)-[(4R)-2,2-dimethyl-1,3-dioxolan-4-yl](hydroxy)ethanenitrile
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?
cyanide + (4R,5S)-2,2,5-trimethyl-1,3-dioxolane-4-carbaldehyde
(2S)-hydroxy-[(4S,5S)-2,2,5-trimethyl-1,3-dioxolan-4-yl]ethanenitrile + (2R)-hydroxy-[(4S,5S)-2,2,5-trimethyl-1,3-dioxolan-4-yl]ethanenitrile
show the reaction diagram
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the natural substrate benzaldehyde is stereoselectively converted to (R)-mandelonitrile. The non-natural substrate (4R,5S)-2,2,5-trimethyl-1,3-dioxolane-4-carbaldehyde is converted to 48.1% (2S)-hydroxy-[(4S,5S)-2,2,5-trimethyl-1,3-dioxolan-4-yl]ethanenitrile and 51.9% (2R)-hydroxy-[(4S,5S)-2,2,5-trimethyl-1,3-dioxolan-4-yl]ethanenitrile
-
?
cyanide + (4R,5S)-2,2-dimethyl-5-phenyl-1,3-dioxolane-4-carbaldehyde
(2S)-[(4S,5S)-2,2-dimethyl-5-phenyl-1,3-dioxolan-4-yl](hydroxy)ethanenitrile + (2R)-[(4S,5S)-2,2-dimethyl-5-phenyl-1,3-dioxolan-4-yl](hydroxy)ethanenitrile
show the reaction diagram
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-
the natural substrate benzaldehyde is stereoselectively converted to (R)-mandelonitrile. The non-natural substrate (4R,5S)-2,2-dimethyl-5-phenyl-1,3-dioxolane-4-carbaldehyde is converted to 52.7% (2S)-[(4S,5S)-2,2-dimethyl-5-phenyl-1,3-dioxolan-4-yl](hydroxy)ethanenitrile and 47.3% (2R)-[(4S,5S)-2,2-dimethyl-5-phenyl-1,3-dioxolan-4-yl](hydroxy)ethanenitrile
-
?
cyanide + (4S)-2,2-dimethyl-1,3-dioxolane-4-carbaldehyde
(2S)-[(4S)-2,2-dimethyl-1,3-dioxolan-4-yl](hydroxy)ethanenitrile + (2R)-[(4S)-2,2-dimethyl-1,3-dioxolan-4-yl](hydroxy)ethanenitrile
show the reaction diagram
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the natural substrate benzaldehyde is stereoselectively converted to (R)-mandelonitrile. The non-natural substrate (4S)-2,2-dimethyl-1,3-dioxolane-4-carbaldehyde is converted to 34.9% (2S)-[(4S)-2,2-dimethyl-1,3-dioxolan-4-yl](hydroxy)ethanenitrile and 65.1% (2R)-[(4S)-2,2-dimethyl-1,3-dioxolan-4-yl](hydroxy)ethanenitrile
-
?
cyanide + (4S,5R)-2,2,5-trimethyl-1,3-dioxolane-4-carbaldehyde
(2S)-hydroxy-[(4R,5R)-2,2,5-trimethyl-1,3-dioxolan-4-yl]ethanenitrile + (2R)-hydroxy-[(4R,5R)-2,2,5-trimethyl-1,3-dioxolan-4-yl]ethanenitrile
show the reaction diagram
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-
the natural substrate benzaldehyde is stereoselectively converted to (R)-mandelonitrile. The non-natural substrate (4S,5R)-2,2,5-trimethyl-1,3-dioxolane-4-carbaldehyde is converted to 35.1% (2S)-hydroxy-[(4R,5R)-2,2,5-trimethyl-1,3-dioxolan-4-yl]ethanenitrile and 64.9% (2R)-hydroxy-[(4R,5R)-2,2,5-trimethyl-1,3-dioxolan-4-yl]ethanenitrile
-
?
cyanide + (4S,5R)-2,2-dimethyl-5-phenyl-1,3-dioxolane-4-carbaldehyde
(2S)-[(4R,5R)-2,2-dimethyl-5-phenyl-1,3-dioxolan-4-yl](hydroxy)ethanenitrile + (2R)-[(4R,5R)-2,2-dimethyl-5-phenyl-1,3-dioxolan-4-yl](hydroxy)ethanenitrile
show the reaction diagram
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the natural substrate benzaldehyde is stereoselectively converted to (R)-mandelonitrile. The non-natural substrate (4S,5R)-2,2-dimethyl-5-phenyl-1,3-dioxolane-4-carbaldehyde is converted to 49.9% (2S)-[(4R,5R)-2,2-dimethyl-5-phenyl-1,3-dioxolan-4-yl](hydroxy)ethanenitrile and 50.1% (2R)-[(4R,5R)-2,2-dimethyl-5-phenyl-1,3-dioxolan-4-yl](hydroxy)ethanenitrile
-
?
cyanide + 1,1'-diformylferrocene
(R,R)-1,1'-bis(cyanohydroxymethyl)ferrocene
show the reaction diagram
-
a bulky organometallic compound, which does not occur in nature. S-hydroxynitrile lyase from Hevea brasieliensis catalyzes the formation of (R,R)-1,1'-bis(cyanohydroxymethyl)ferrocene at high yield and stereochemical purity
obtained at high yield and stereochemical purity
-
?
cyanide + 1,3-benzodioxole-5-carbaldehyde
(2S)-1,3-benzodioxol-5-yl(hydroxy)acetonitrile
show the reaction diagram
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-
86% enantiomeric excess
-
?
cyanide + 1,4-dioxaspiro[4.5]decane-2-carbaldehyde
(S)-2-hydroxy-2-((R)-1,4-dioxaspiro[4.5]decan-2-yl)acetonitrile + (R)-2-hydroxy-2-((R)-1,4-dioxaspiro[4.5]decan-2-yl)acetonitrile + (S)-2-hydroxy-2-((S)-1,4-dioxaspiro[4.5]decan-2-yl)acetonitrile + (R)-2-hydroxy-2-((S)-1,4-dioxaspiro[4.5]decan-2-yl)acetonitrile
show the reaction diagram
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-
the natural substrate benzaldehyde is stereoselectively converted to (R)-mandelonitrile. The non-natural substrate 1,4-dioxaspiro[4.5]decane-2-carbaldehyde is converted to 16.9% (2S)-(2R)-1,4-dioxaspiro[4.5]dec-2-yl(hydroxy)ethanenitrile, 33.0% (2R)-(2R)-1,4-dioxaspiro[4.5]dec-2-yl(hydroxy)ethanenitrile, 18.3% (2S)-(2S)-1,4-dioxaspiro[4.5]dec-2-yl(hydroxy)ethanenitrile and 31.8% (2R)-(2S)-1,4-dioxaspiro[4.5]dec-2-yl(hydroxy)ethanenitrile
-
?
cyanide + 1-phenylethanone
(2S)-2-hydroxy-2-phenylpropanenitrile
show the reaction diagram
-
wild-type enzyme: 87% enantiomeric excess, 13% conversion rate
-
-
?
cyanide + 1-phenylpropan-2-one
(2S)-2-hydroxy-2-methyl-3-phenylpropanenitrile
show the reaction diagram
-
wild-type enzyme: 97% enantiomeric excess, 82% conversion rate
-
-
?
cyanide + 2,2-dimethylpropanal
(2S)-2-hydroxy-3,3-dimethylbutanenitrile
show the reaction diagram
cyanide + 2,4-dimethylbenzaldehyde
(2S)-(2,4-dimethylphenyl)(hydroxy)ethanenitrile
show the reaction diagram
-
65% yield, 82% enantiomeric excess
-
?
cyanide + 2-bromobenzaldehyde
(2S)-(2-bromophenyl)(hydroxy)ethanenitrile
show the reaction diagram
-
-
wild-type enzyme: 96% enantiomeric excess, 96% conversion rate
-
?
cyanide + 2-chlorobenzaldehyde
(2S)-(2-chlorophenyl)(hydroxy)acetonitrile
show the reaction diagram
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92% enantiomeric excess
-
?
cyanide + 2-chlorobenzaldehyde
(2S)-(2-chlorophenyl)(hydroxy)ethanenitrile
show the reaction diagram
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wild-type enzyme: 98% enantiomeric excess, 96% conversion rate
-
?
cyanide + 2-hydroxybenzaldehyde
(2S)-(2-hydroxyphenyl)(hydroxy)ethanenitrile
show the reaction diagram
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wild-type enzyme: 91% enantiomeric excess, 47% conversion rate
-
?
cyanide + 2-methoxybenzaldehyde
(2S)-2-hydroxy-2-(2-methoxyphenyl)acetonitrile
show the reaction diagram
-
77% enantiomeric excess
-
?
cyanide + 2-methylbenzaldehyde
(2S)-(2-methylphenyl)(hydroxy)ethanenitrile
show the reaction diagram
-
76% yield, 47% enantiomeric excess
-
?
cyanide + 2-methylpropanal
(2S)-2-hydroxy-3-methylbutanenitrile
show the reaction diagram
cyanide + 3,3-dimethylbutan-2-one
(2S)-2-hydroxy-2,3,3-trimethylbutanenitrile
show the reaction diagram
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-
78% enantiomeric excess
-
?
cyanide + 3-(4-hydroxyphenyl)propanal
(2S)-2-hydroxy-4-(4-hydroxyphenyl)butanenitrile
show the reaction diagram
-
-
wild-type enzyme: 67% enantiomeric excess, 90% conversion rate
-
?
cyanide + 3-hydroxybenzaldehyde
(2S)-(3-hydroxyphenyl)(hydroxy)ethanenitrile
show the reaction diagram
-
-
wild-type enzyme: 97% enantiomeric excess, 88% conversion rate
-
?
cyanide + 3-methoxybenzaldehyde
(2S)-(3-methoxyphenyl)(hydroxy)ethanenitrile
show the reaction diagram
-
67% yield, 76% enantiomeric excess
-
?
cyanide + 3-methoxybenzaldehyde
(2S)-2-hydroxy-2-(3-methoxyphenyl)acetonitrile
show the reaction diagram
-
99% enantiomeric excess
-
?
cyanide + 3-methylbenzaldehyde
(2S)-(3-methylphenyl)(hydroxy)ethanenitrile
show the reaction diagram
-
76% yield, 76% enantiomeric excess
-
?
cyanide + 3-phenoxybenzaldehyde
(2S)-2-hydroxy-2-(3-phenoxyphenyl)acetonitrile
show the reaction diagram
-
99% enantiomeric excess
-
?
cyanide + 3-phenoxybenzaldehyde
(2S)-hydroxy(3-phenoxyphenyl)acetonitrile
show the reaction diagram
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-
20% enantiomeric excess
-
?
cyanide + 3-phenoxybenzaldehyde
(S)-3-phenoxybenzaldehyde cyanohydrin
show the reaction diagram
-
reaction in a high-pH two-phase system
97% enantiomeric excess
-
r
cyanide + 3-phenylpropanal
(2S)-2-hydroxy-4-phenylbutanenitrile
show the reaction diagram
cyanide + 3-tetrahydrothiophenone
(S)-3-hydroxytetrahydrothiophene-3-carbonitrile
show the reaction diagram
-
-
-
-
?
cyanide + 4-hydroxybenzaldehyde
(2S)-(4-hydroxyphenyl)(hydroxy)ethanenitrile
show the reaction diagram
-
-
wild-type enzyme: 94% enantiomeric excess, 51% conversion rate
-
?
cyanide + 4-methoxybenzaldehyde
(2S)-(4-methoxyphenyl)(hydroxy)ethanenitrile
show the reaction diagram
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-
wild-type enzyme: 99% enantiomeric excess, 79% conversion rate
-
?
cyanide + 4-methoxybenzaldehyde
(2S)-2-hydroxy-2-(4-methoxyphenyl)acetonitrile
show the reaction diagram
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95% enantiomeric excess
-
?
cyanide + 4-methoxybenzaldehyde
(2S)-4-methoxymandelonitrile
show the reaction diagram
-
-
-
-
?
cyanide + 4-methoxybenzaldehyde
(2S)-hydroxy(4-methoxyphenyl)acetonitrile
show the reaction diagram
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98% enantiomeric excess
-
?
cyanide + 4-methoxycyclohex-3-ene-1-carbaldehyde
(2S)-hydroxy[4-(methoxy)cyclohex-3-en-1-yl]ethanenitrile
show the reaction diagram
-
-
-
-
r
cyanide + 4-methylbenzaldehyde
(2S)-(4-methylphenyl)(hydroxy)ethanenitrile
show the reaction diagram
-
-
wild-type enzyme: 99% enantiomeric excess, 50% conversion rate
-
?
cyanide + 4-methylpentan-2-one
(2S)-2-hydroxy-2,4-dimethylpentanenitrile
show the reaction diagram
-
-
28% enantiomeric excess
-
?
cyanide + 4-oxocyclohexanecarbaldehyde
(2S)-hydroxy(4-oxocyclohexyl)ethanenitrile
show the reaction diagram
-
-
-
-
r
cyanide + 4-phenoxybenzaldehyde
(2S)-(4-phenoxyphenyl)(hydroxy)ethanenitrile
show the reaction diagram
-
-
wild-type enzyme: 96% enantiomeric excess, 47% conversion rate
-
?
cyanide + 4-phenylbutan-2-one
(2S)-2-hydroxy-2-methyl-4-phenylbutanenitrile
show the reaction diagram
-
wild-type enzyme: 49% enantiomeric excess, 36% conversion rate
-
-
?
cyanide + 4-[(trimethylsilyl)oxy]cyclohex-3-ene-1-carbaldehyde
(2S)-hydroxy[4-((trimethylsilyl)oxy)cyclohex-3-en-1-yl]ethanenitrile
show the reaction diagram
-
-
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-
r
cyanide + 6-methylhept-5-en-2-one
(2S)-2-hydroxy-2,6-dimethylhept-5-enenitrile
show the reaction diagram
-
wild-type enzyme: 61% enantiomeric excess, 78% conversion rate
-
-
?
cyanide + benzaldehyde
(2S)-2-hydroxy-2-phenylacetonitrile
show the reaction diagram
-
i.e. (S)-mandelonitrile, more than 99% enantiomeric excess
-
?
cyanide + benzaldehyde
(S)-mandelonitrile
show the reaction diagram
cyanide + butan-2-one
(2S)-2-hydroxy-2-methylbutanenitrile
show the reaction diagram
-
-
18% enantiomeric excess
-
?
cyanide + butanal
(2S)-2-hydroxypentanenitrile
show the reaction diagram
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-
80% enantiomeric excess
-
?
cyanide + cinnamaldehyde
(2S)-2-hydroxy-4-phenyl-(E)-but-3-enenitrile
show the reaction diagram
-
95% enantiomeric excess
-
?
cyanide + cyclohex-3-ene-1-carbaldehyde
(2S)-2-(cyclohex-3-enyl)-2-hydroxyacetonitrile
show the reaction diagram
-
99% enantiomeric excess
-
?
cyanide + cyclohexanecarbaldehyde
(2S)-2-cyclohexyl-2-hydroxyacetonitrile
show the reaction diagram
-
99% enantiomeric excess
-
?
cyanide + cyclohexanecarbaldehyde
(2S)-cyclohexyl(hydroxy)acetonitrile
show the reaction diagram
-
-
92% enantiomeric excess
-
?
cyanide + decanal
(2S)-2-hydroxyundecanenitrile
show the reaction diagram
-
-
wild-type enzyme: 78% enantiomeric excess, 65% conversion rate
-
?
cyanide + dodecanal
(2S)-2-hydroxytridecanenitrile
show the reaction diagram
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-
wild-type enzyme: 71% enantiomeric excess, 80% conversion rate
-
?
cyanide + ferrocenecarboxaldehyde
(R)-(cyanohydroxymethyl)ferrocene
show the reaction diagram
-
i.e. bis(cyclopentadienyl)iron, a bulky organometallic compound, which does not occur in nature. S-hydroxynitrile lyase from Hevea brasiliensis catalyzes the formation of (R)-(cyanohydroxymethyl)ferrocene at high yield and stereochemical purity
obtained at high yield and stereochemical purity
-
?
cyanide + heptan-2-one
(2S)-2-hydroxy-2-methylheptanenitrile
show the reaction diagram
-
-
92% enantiomeric excess
-
?
cyanide + heptan-3-one
(2S)-2-ethyl-2-hydroxyhexanenitrile
show the reaction diagram
-
wild-type enzyme: 46% enantiomeric excess, 14% conversion rate
-
-
?
cyanide + hexan-2-one
(2S)-2-hydroxy-2-methylhexanenitrile
show the reaction diagram
-
-
80% enantiomeric excess
-
?
cyanide + hexanal
2-hydroxyheptanenitrile
show the reaction diagram
-
-
84% enantiomeric excess
-
?
cyanide + nonanal
(2S)-2-hydroxydecanenitrile
show the reaction diagram
-
-
wild-type enzyme: 80% enantiomeric excess, 99% conversion rate
-
?
cyanide + nonanal
2-hydroxydecanenitrile
show the reaction diagram
-
-
85% enantiomeric excess
-
?
cyanide + octan-3-one
(2S)-2-ethyl-2-hydroxyheptanenitrile
show the reaction diagram
-
wild-type enzyme: 61% enantiomeric excess, 24% conversion rate
-
-
?
cyanide + octanal
(2S)-2-hydroxynonanenitrile
show the reaction diagram
-
-
wild-type enzyme: 79% enantiomeric excess, 96% conversion rate
-
?
cyanide + pentan-2-one
(2S)-2-hydroxy-2-methylpentanenitrile
show the reaction diagram
-
-
69% enantiomeric excess
-
?
cyanide + pentanal
(2S)-2-hydroxyhexanenitrile
show the reaction diagram
-
-
91% enantiomeric excess
-
?
cyanide + phenylacetaldehyde
(2S)-2-hydroxy-3-phenylpropanenitrile
show the reaction diagram
-
99% enantiomeric excess
-
?
cyanide + phenylacetaldehyde
2-hydroxy-3-phenylpropanenitrile
show the reaction diagram
-
-
wild-type enzyme: 98% enantiomeric excess, 99% conversion rate
-
?
cyanide + piperonal
?
show the reaction diagram
-
47% yield, 74% enantiomeric excess
-
?
cyanide + prop-2-enal
(2S)-2-hydroxy-3-methylbutanenitrile
show the reaction diagram
-
-
-
-
?
cyanide + prop-2-enal
(2S)-2-hydroxybut-3-enenitrile
show the reaction diagram
cyanide + prop-2-enal
2-hydroxybut-3-enenitrile
show the reaction diagram
-
-
84% enantiomeric excess
-
?
cyanide + propanal
(2S)-2-hydroxybutanenitrile
show the reaction diagram
-
-
91% enantiomeric excess
-
?
cyanide + propanal
(2S)-2-hydroxypentanenitrile
show the reaction diagram
-
-
-
-
?
cyanide + tetrahydro-2H-pyran-2-carbaldehyde
(2S)-hydroxy-[(2R)-tetrahydro-2H-pyran-2-yl]ethanenitrile + (2R)-hydroxy-[(2R)-tetrahydro-2H-pyran-2-yl]ethanenitrile + (2S)-hydroxy-[(2S)-tetrahydro-2H-pyran-2-yl]ethanenitrile + (2R)-hydroxy-[(2S)-tetrahydro-2H-pyran-2-yl]ethanenitrile
show the reaction diagram
-
-
the natural substrate benzaldehyde is stereoselectively converted to (R)-mandelonitrile. The non-natural substrate tetrahydro-2H-pyran-2-carbaldehyde is converted to 5.4% (2S)-hydroxy-[(2R)-tetrahydro-2H-pyran-2-yl]ethanenitrile, 45.9% (2R)-hydroxy-[(2R)-tetrahydro-2H-pyran-2-yl]ethanenitrile, 3.9% (2S)-hydroxy[(2S)-tetrahydro-2H-pyran-2-yl]ethanenitrile and 44.9% (2R)-hydroxy[(2S)-tetrahydro-2H-pyran-2-yl]ethanenitrile
-
?
cyanide + tetrahydrofuran-2-carbaldehyde
(2S)-hydroxy-[(2R)-tetrahydrofuran-2-yl]ethanenitrile + (2R)-hydroxy-[(2R)-tetrahydrofuran-2-yl]ethanenitrile + (2S)-hydroxy-[(2S)-tetrahydrofuran-2-yl]ethanenitrile + (2R)-hydroxy-[(2S)-tetrahydrofuran-2-yl]ethanenitrile
show the reaction diagram
-
-
the natural substrate benzaldehyde is stereoselectively converted to (R)-mandelonitrile. The non-natural substrate tetrahydrofuran-2-carbaldehyde is converted to 17.1% (2S)-hydroxy-[(2R)-tetrahydrofuran-2-yl]ethanenitrile, 32.9% (2R)-hydroxy-[(2R)-tetrahydrofuran-2-yl]ethanenitrile, 18.9% (2S)-hydroxy[(2S)-tetrahydrofuran-2-yl]ethanenitrile and 31.1% (2R)-hydroxy[(2S)-tetrahydrofuran-2-yl]ethanenitrile
-
?
cyanide + thiophene-2-carbaldehyde
(2S)-hydroxy(thiophen-2-yl)ethanenitrile
show the reaction diagram
-
-
-
-
?
cyclohexanone cyanohydrin
?
show the reaction diagram
-
-
-
-
?
DL-mandelonitrile
benzaldehyde + HCN
show the reaction diagram
-
-
-
?
ferrocenyl aldehyde + HCN
ferrocenyl-cyanohydrin
show the reaction diagram
-
-
-
-
?
furan-3-carbaldehyde + HCN
(2S)-hydroxy(furan-3-yl)ethanenitrile
show the reaction diagram
-
-
92% enantiomeric excess
-
?
HCN + (2E)-oct-2-enal
(2S,3E)-2-hydroxynon-3-enenitrile
show the reaction diagram
-
-
-
?
HCN + (benzyloxy)acetaldehyde
3-(benzyloxy)-(2S)-2-hydroxy-propanenitrile + 3-(benzyloxy)-(2R)-2-hydroxy-propanenitrile
show the reaction diagram
-
-
50% 3-(benzyloxy)-(2S)-2-hydroxy-propanenitrile and 50% 3-(benzyloxy)-(2R)-2-hydroxy-propanenitrile
?
HCN + (E)-2-butenal
(3E)-2-hydroxypent-3-enenitrile
show the reaction diagram
-
-
-
?
HCN + 1,1'-diformylferrocene
(R,R)-1,1-bis(cyanohydroxymethyl)ferrocene
show the reaction diagram
-
-
-
?
HCN + 2,2-dimethylpropanal
2-hydroxy-3,3-dimethylbutyronitrile
show the reaction diagram
-
-
-
?
HCN + 2-allyloxybutanal
3-allyloxy-2-hydroxypentanenitrile
show the reaction diagram
-
-
-
?
HCN + 2-allyloxyheptanal
(2R,3RS)-3-allyloxy-2-hydroxyoctanenitrile
show the reaction diagram
-
-
-
?
HCN + 2-allyloxyhexanal
(2R,3RS)-3-allyloxy-2-hydroxyheptanenitrile
show the reaction diagram
-
-
-
?
HCN + 2-allyloxypentanal
(2R,3RS)-3-allyloxy-2-hydroxyhexanenitrile
show the reaction diagram
-
-
-
?
HCN + 2-allyloxypropanal
3-allyloxy-2-hydroxybutanenitrile
show the reaction diagram
-
-
-
?
HCN + 2-benzyloxypropanal
3-benzyloxy-2-hydroxybutanenitrile
show the reaction diagram
-
-
-
?
HCN + 2-chlorobenzaldehyde
(2-chlorophenyl)(hydroxy)acetonitrile
show the reaction diagram
-
-
-
?
HCN + 2-hexenal
2-hydroxyhept-3-enenitrile
show the reaction diagram
-
-
-
?
HCN + 2-methoxymethoxypropanal
3-methoxymethoxy-2-hydroxybutanenitrile
show the reaction diagram
-
-
-
?
HCN + 2-methylallyloxyacetaldehyde
3-(2-methylallyloxy)-2-hydroxypropionitrile
show the reaction diagram
-
-
-
?
HCN + 2-methyldihydrofuran
3-hydroxy-2-methyltetrahydrofuran-3-carbonitrile
show the reaction diagram
-
analysis of diastereomeric distribution of the products, depending on different reaction conditions such as pH, reaction time, and solvent properties
-
?
HCN + 2-methyldihydrothiophen-3(2H)-one
3-hydroxy-2-methyltetrahydrothiophen-3-carbonitrile
show the reaction diagram
-
analysis of diastereomeric distribution of the products, depending on different reaction conditions such as pH, reaction time, and solvent properties
-
?
HCN + 2-naphthaldehyde
(2S)-2-hydroxynaphthalen-2-yl-acetonitrile + (2R)-2-hydroxynaphthalen-2-yl-acetonitrile
show the reaction diagram
-
-
83% (2S)-2-hydroxynaphthalen-2-yl-acetonitrile and 17% (2R)-2-hydroxynaphthalen-2-yl-acetonitrile
?
HCN + 2-naphthylacetaldehyde
(2S)-2-hydroxy-3-naphthalen-1-yl-propionitrile + (2R)-2-hydroxy-3-naphthalen-1-yl-propionitrile
show the reaction diagram
-
-
84.3% (2S)-2-hydroxy-3-naphthalen-1-yl-propionitrile and 15.6% (2R)-2-hydroxy-3-naphthalen-1-yl-propionitrile
?
HCN + 2-propenal
2-hydroxybut-3-enenitrile
show the reaction diagram
-
-
-
?
HCN + 3-furaldehyde
(2R)-3-furyl(hydroxy)acetonitrile
show the reaction diagram
-
-
-
?
HCN + 3-furylcarbaldehyde
hydroxy(fur-3yl)acetonitrile
show the reaction diagram
-
-
-
?
HCN + 3-phenoxybenzaldehyde
(2S)-hydroxy(3-phenoxyphenyl)acetonitrile
show the reaction diagram
-
-
-
?
HCN + 3-phenoxypropanal
(2S)-2-hydroxy-4-phenoxybutanenitrile + (2S)-2-hydroxy-4-phenoxybutanenitrile
show the reaction diagram
-
-
95.8% (2S)-2-hydroxy-4-phenoxybutanenitrile and 4.2% (2R)-2-hydroxy-4-phenoxybutanenitrile
?
HCN + 3-phenylpropionaldehyde
(2S)-2-hydroxy-4-phenylbutanenitrile
show the reaction diagram
HCN + 4-methoxybenzaldehyde
(4-methoxyphenyl) (hydroxy)acetonitrile
show the reaction diagram
-
-
-
?
HCN + acrolein
(2S)-2-hydroxybut-3-enenitrile
show the reaction diagram
HCN + allyloxy-2-hydroxypropionitrile
3-allyloxy-2-hydroxypropionitrile
show the reaction diagram
-
-
-
?
HCN + benzaldehyde
(R)-mandelonitrile
show the reaction diagram
HCN + benzaldehyde
(S)-mandelonitrile
show the reaction diagram
HCN + benzene-1,2,4-tricarbaldehyde
?
show the reaction diagram
-
-
-
?
HCN + benzyloxyacetaldehyde
3-benzyloxy-2-hydroxypropionitrile
show the reaction diagram
-
-
-
?
HCN + cyclohexanecarbaldehyde
(2R)-cyclohexyl(hydroxy)acetonitrile
show the reaction diagram
-
-
-
?
HCN + decanal
(S)-2-hydroxyundecanenitrile
show the reaction diagram
-
reaction in a two phase solvent system aqueous buffer and ionic liquid. Compared to the use of organic solvents as the nonaqueous phase, the reaction rate is significantly increased whereas the enantioselectivity remains good
-
-
?
HCN + dodecanal
(S)-2-hydroxytridecanenitrile
show the reaction diagram
-
reaction in a two phase solvent system aqueous buffer and ionic liquid. Compared to the use of organic solvents as the nonaqueous phase, the reaction rate is significantly increased whereas the enantioselectivity remains good
-
-
?
HCN + ferrocene aldehyde
?
show the reaction diagram
-
-
-
?
HCN + formylferrocene
(R)-(cyanohydroxymethyl)ferrocene
show the reaction diagram
-
-
-
?
HCN + furaldehyde
(2R)-furan-2-yl(hydroxy)ethanenitrile
show the reaction diagram
-
enzyme encapsulated in sol-gel matrix
-
-
?
HCN + hexanal
(S)-2-hydroxyoctanenitrile
show the reaction diagram
-
enzyme encapsulated in sol-gel matrix
-
-
?
HCN + isobutyraldehyde
2-hydroxy-3-methylbutyronitrile
show the reaction diagram
-
-
-
?
HCN + m-phenoxybenzaldehyde
(S)-hydroxy-(3-phenoxy-phenyl)acetonitrile
show the reaction diagram
-
enzyme encapsulated in sol-gel matrix
-
-
?
HCN + methoxymethoxyacetaldehyde
2-hydroxy-3-methoxymethoxypropionitrile
show the reaction diagram
-
-
-
?
HCN + methyl isopropyl ketone
(S)-2-hydroxy-2,3-dimethylbutanenitrile
show the reaction diagram
-
enzyme encapsulated in sol-gel matrix
-
-
?
HCN + pentanal
2-hydroxyhexanenitrile
show the reaction diagram
-
-
-
?
HCN + phenylacetaldehyde
(2R)-2-hydroxy-3-phenylpropanenitrile
show the reaction diagram
-
-
-
?
HCN + propanal
2-hydroxybutanenitrile
show the reaction diagram
-
-
-
?
HCN + rac-2-methyl-3-phenylpropionaldehyde
(2S,3R)-2-hydroxy-3-methyl-4-phenylbutyronitrile
show the reaction diagram
-
-
wild-type and mutant enzymes Y128Y, W128L, W128C, W128A are (S)-selective
-
?
HCN + rac-2-phenylbutyraldehyde
2-hydroxy-3-phenylpentanenitrile
show the reaction diagram
-
-
diastereomer composition. (2R,3R): 0.5% (wild-type), 0.7% (mutant W128Y), 0.8% (mutant W128L), 0.4% (mutant W128L), 0.6% (mutant W128C), 0% (mutant W128A). (2R,3S): 4.5% (wild-type), 4.6% (mutant W128Y), 24.3% (mutant W128L), 23.4% (mutant W128C), 34.1% (mutant W128A), 35.5% (mutant W128V). (2S,3S): 45.8% (wild-type), 45.6% (mutant W128Y), 25.1% (mutant W128L), 26.1% (mutant W128C), 15.0% (mutant W128A), 13.7% (mutant W128V). (2S,3R): 49.2% (wild-type), 49.1% (mutant W128Y), 49.8% (mutant W128L), 50.1% (mutant W128C), 50.3% (mutant W128A), 50.8% (mutant W128V)
-
?
HCN + rac-2-phenylpropionaldehyde
2-hydroxy-3-phenylbutyronitrile
show the reaction diagram
-
-
diastereomer composition. (2R,3R): 0.1% (wild-type), 0.2% (mutant W128Y), 0.3% (mutant W128L), 0.5% (mutant W128C), 1% (mutant W128A). (2R,3S): 5.2% (wild-type), 24.4% (mutant W128Y), 37.5% (mutant W128L), 43.4% (mutant W128C), 46.2% mutant (W128A). (2S,3S): 44.4% (wild-type), 25.4% (mutant W128Y), 12.2% (mutant W128C), 6.1% (mutant W128L), 3.4% (mutant W128A). (2S,3R): 50.3% (wild-type), 50.2% (mutant W128Y), 50.0% (mutant W128L), 50.0% (mutant W128C), 49.4% (mutant W128A)
-
?
HCN + rac-3-phenylbutyraldehyde
(2S,3R)-2-hydroxy-4-phenylpentanenitrile
show the reaction diagram
-
-
wild-type and mutant enzymes Y128Y, W128L, W128C, W128A are (S)-selective
-
?
HCN + tetrahydro-2H-3-pyranone
(3R)-3-hydroxytetrahydro-2H-pyran-3-carbonitrile
show the reaction diagram
-
-
enantiomeric excess at pH 4.75 is 48.3%
?
HCN + tetrahydro-3-furanone
(3R)-3-hydroxytetrahydrofuran-3-carbonitrile
show the reaction diagram
-
-
81% enantiomeric excess
?
HCN + thiophen-2-carbaldehyde
hydroxy(thien-2-yl)acetonitrile
show the reaction diagram
-
-
-
?
HCN + thiophen-3-carbaldehyde
hydroxy(thien-3-yl)acetonitrile
show the reaction diagram
-
-
-
?
HCN + undecanal
(S)-2-hydroxydodecanenitrile
show the reaction diagram
-
reaction in a two phase solvent system aqueous buffer and ionic liquid. Compared to the use of organic solvents as the nonaqueous phase, the reaction rate is significantly increased whereas the enantioselectivity remains good
-
-
?
hexanal cyanohydrin
hexanal + HCN
show the reaction diagram
Isobutyraldehyde + cyanide
2-Hydroxy-3-methylbutyronitrile
show the reaction diagram
-
-
-
-
-
lactonitrile
?
show the reaction diagram
-
poor substrate
-
-
?
m-phenoxybenzaldehyde cyanohydrin
m-phenoxybenzaldehyde + cyanide
show the reaction diagram
-
2-phenoxybenzaldehyde cyanohydrin is converted with lower activity than (S)-mandelonitrile and cyclohexanone cyanohydrile
-
-
?
m-phenoxybenzaldehyde cyanohydrin
m-phenoxybenzaldehyde + HCN
show the reaction diagram
mandelonitrile
benzaldehyde + HCN
show the reaction diagram
mandelonitrile
HCN + benzaldehyde
show the reaction diagram
n-Butyraldehyde + cyanide
(S)-2-Hydroxyvaleronitrile
show the reaction diagram
-
-
-
-
nitromethane + benzaldehyde
(S)-2-nitro-1-phenylethanol
show the reaction diagram
-
-
-
-
r
p-hydroxymandelonitrile
?
show the reaction diagram
-
phydroxymandelonitrile is converted with lower activity than (S)-mandelonitrile and cyclohexanone cyanohydrile
-
-
?
propionaldehyde cyanohydrin
propionaldehyde + cyanide
show the reaction diagram
-
poor substrate
-
-
?
rac-mandelonitrile
HCN + benzaldehyde
show the reaction diagram
-
-
-
?
thiophene-2-carbaldehyde + HCN
(2S)-hydroxy(thiophen-2-yl)ethanenitrile
show the reaction diagram
-
-
96% enantiomeric excess
-
?
thiophene-3-carbaldehyde + HCN
(2S)-hydroxy(thiophen-3-yl)ethanenitrile
show the reaction diagram
-
-
98% enantiomeric excess
-
?
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
(2S)-2-hydroxy-2-methylbutanenitrile
cyanide + butan-2-one
show the reaction diagram
P52705
the liberation of HCN serves as a defense mechanism against herbivores and microbial attack in plants
-
-
?
2-hydroxy-2-methylpropanenitrile
acetone + HCN
show the reaction diagram
2-hydroxy-2-methylpropanenitrile
cyanide + acetone
show the reaction diagram
2-hydroxy-2-methylpropanenitrile
HCN + acetone
show the reaction diagram
Acetone cyanhydrin
Cyanide + acetone
show the reaction diagram
acetone cyanohydrin
cyanide + acetone
show the reaction diagram
-
-
-
-
?
mandelonitrile
HCN + benzaldehyde
show the reaction diagram
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-butyl-3-methylimidazolium tetrahydroborate
-
2-6% v/v, inactivates
1-hexyl-3-methylimidazolium tetrahydroborate
-
2-6% v/v, inactivates
1-pentyl-3-methylimidazolium tetrahydroborate
-
2-6% v/v, inactivates
1-propyl-3-methylimidazolium tetrahydroborate
-
2-6% v/v, inactivates
2-methyl-2-butanol
-
-
2-Methylbutyraldehyde
-
-
4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride
-
-
acetaldehyde
-
-
acetate
acetone
Ag+
1 mM, 12% residual activity
benzaldehyde
butanol
-
-
Chlorobutanol
-
-
diethyl dicarbonate
-
3 MM, 93% inhibition
diisopropyl fluorophosphate
ethanol
-
-
formaldehyde
-
-
HCN
-
shows S-linear I-parabolic mixed-type inhibition
hexafluoroacetone
-
-
isobutanol
-
-
Isobutyraldehyde
-
-
Mandelonitrile
-
-
methanol
-
-
methyl ethyl ketone
Octanol
-
-
p-chloromercurybenzoate
-
-
Pentanol
-
-
Phenol
Phenylmethyl sulfonylfluoride
1 mM, 27% residual activity
phenylmethylsulfonyl fluoride
-
-
Propanol
-
-
propionaldehyde
-
-
rhodanide
-
very strong competitive inhibitor
Thiocyanate
trichloracetaldehyde
-
-
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
dibutyl ether
-
best solvent for HCN concentrations around 300 mM HCN concentration
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.6
(S)-2-nitro-1-phenylethanol
-
in 50 mM phosphate buffer pH 6.0, at 22C; in 50 mM phosphate buffer pH 6.0, at 25C
1.2 - 30
(S)-Mandelonitrile
5.2
2,4-dimethylbenzaldehyde
pH 6.0, 25C
67 - 150
2-hydroxy-2-methylpropanenitrile
0.8
2-Methyl-2-hydroxybutyronitrile
-
-
21.7
2-Methylbenzaldehyde
pH 6.0, 25C
161.5
3,3-dimethyl-2-butanone
-
pH 5.4, 40C
9.8
3-Methoxybenzaldehyde
pH 6.0, 25C
14.4
3-methylbenzaldehyde
pH 6.0, 25C
0.7 - 174
acetone cyanohydrin
61.5
acetyltrimethylsilane
-
pH 5.4, 40C
2 - 27.9
benzaldehyde
0.93
CN-
-
-
179
cyanide
-
citrate buffer (50 mM, pH 4.0), at 25C; pH 7.5
1.1 - 1.4
Mandelonitrile
15.7
piperonal
pH 6.0, 25C
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.83 - 41.8
(S)-Mandelonitrile
18.5
2,4-dimethylbenzaldehyde
Baliospermum montanum
D1MX73
pH 6.0, 25C
53.6
2-Methylbenzaldehyde
Baliospermum montanum
D1MX73
pH 6.0, 25C
28.1
3-Methoxybenzaldehyde
Baliospermum montanum
D1MX73
pH 6.0, 25C
49.2
3-methylbenzaldehyde
Baliospermum montanum
D1MX73
pH 6.0, 25C
26.6 - 96.6
benzaldehyde
91.4
piperonal
Baliospermum montanum
D1MX73
pH 6.0, 25C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.55 - 15.83
(S)-Mandelonitrile
3.56
2,4-dimethylbenzaldehyde
Baliospermum montanum
D1MX73
pH 6.0, 25C
128269
2.47
2-Methylbenzaldehyde
Baliospermum montanum
D1MX73
pH 6.0, 25C
3593
2.87
3-Methoxybenzaldehyde
Baliospermum montanum
D1MX73
pH 6.0, 25C
1856
3.41
3-methylbenzaldehyde
Baliospermum montanum
D1MX73
pH 6.0, 25C
2710
3.44 - 6.75
benzaldehyde
5.82
piperonal
Baliospermum montanum
D1MX73
pH 6.0, 25C
13828
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.37
benzaldehyde
-
in 50 mM phosphate buffer pH 6.0, at 22C; in 50 mM phosphate buffer pH 6.0, at 25C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.7
-
recombinant mutant enzyme H103L expressed in an Escherichia coli lysate (WakoPURE system), using benzaldehyde as substrate, pH and temperature not specified in the publication
1.02
-
C-terminal His-tagged recombinant mutant enzyme H103L expressed in a RTS 100 wheat germ cell-free translation system, using benzaldehyde as substrate, pH and temperature not specified in the publication
1.1
-
recombinant mutant enzyme H103L expressed in Pichia pastoris, using benzaldehyde as substrate, pH and temperature not specified in the publication
1.55
-
C-terminal His-tagged recombinant wild type enzyme expressed in a RTS 100 wheat germ cell-free translation system, using benzaldehyde as substrate, pH and temperature not specified in the publication
1.73
-
recombinant wild type enzyme expressed in Escherichia coli JM109 cells, using benzaldehyde as substrate, pH and temperature not specified in the publication
1.97
-
untagged recombinant wild type enzyme expressed in a RTS 100 wheat germ cell-free translation system, using benzaldehyde as substrate, pH and temperature not specified in the publication
1.98
-
untagged recombinant mutant enzyme H103L expressed in a RTS 100 wheat germ cell-free translation system, using benzaldehyde as substrate, pH and temperature not specified in the publication
2.03
-
recombinant wild type enzyme expressed in Leishmania tarentolae, using benzaldehyde as substrate, pH and temperature not specified in the publication
2.1
-
recombinant wild type enzyme expressed in Pichia pastoris, using benzaldehyde as substrate, pH and temperature not specified in the publication
2.18
-
recombinant mutant enzyme H103L expressed in Leishmania tarentolae, using benzaldehyde as substrate, pH and temperature not specified in the publication
29.2
-
recombinant mutant enzyme H103L expressed in Escherichia coli JM109 cells, using benzaldehyde as substrate, pH and temperature not specified in the publication
35.6
-
wild-type enzyme
57.2
-
mutant enzyme G113S
92
-
wild-type enzyme from leaves
240
-
native enzyme
250
-
recombinant enzyme
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.8
-
conversion of formylferrocene
5.2
-
assay at
5.3 - 5.7
-
substrate: acetone cyanohydrin
5.6
-
wild-type enzyme
7.5
-
the maximal activity of HNL in (S)-2-nitro-1-phenylethanol cleavage is at/above pH 7.5
8
-
synthesis of (S)-3-phenoxybenzaldehyde cyanohydrin
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 5.8
-
pH 4: about 40% of maximal activity, pH 5.8: optimum
4.5 - 6.5
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pH 4.5: about 65% of maximal activity of mutant enzyme G113S, about 60% of maximal activity of wild-type enzyme, pH 6.5: about 50% of maximal activity of mutant enzyme G113S, about 55% of maximal activity of wild-type enzyme
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
23
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
-5 - 25
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
2% of the activity of leaves
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
29210
-
electrospray mass spectrometry
30000
recombinant enzyme, SDS-PAGE
50100
-
gel filtration
60000 - 65000
gel filtration
92000 - 124000
-
-
92000
-
gel filtration
100000 - 105000
-
gel filtration
102000
-
gel filtration
124000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 30000
homotetramer
homotrimer
polymer
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x * 28000-30000
tetramer
trimer
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3 * 29481, calculation from sequence
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
no glycoprotein
-
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure of the hydroxynitrile lyase at 1.9 A resolution. The structure belongs to the alpha/beta hydrolase superfamily. Its active site is deeply buried inside the protein, and connected to the outside by a narrow tunnel. The catalytic triade consists of residues Ser80, His235 and Asp207. By analogy with other alpha/beta hydrolases, the oxyanion hole is formed by the mainchain-NH of Cys81 and by the side-chains of Cys81 and Thr11
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crystals obtained by sitting drop vapor diffusion, space group C222(1), unit cell dimensions a = 47.29 A, b = 106.66 A, c = 128.16 A
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enzyme in complex with acetone, hexafluoroacetone, trichloroacetaldehyde or rhodanine, hanging-drop vapour diffusion method, space group C222(1)
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hanging drop vapor diffusion method, freeze-quench method to prepare crystals of the complex of the enzyme with acetone cyanohydrin, complex of mutant K236L enzyme with acetone, structure of the K236L with acetone cyanohydrin-acetone cyanohydrin shows the substrate in a different orientation from the wild-type complex
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hanging drop vapor diffusion. crystal structures of hydroxynitrile lyase: one native and three complexes with acetone, isopropyl alcohol, and thiocyanate
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hanging-drop vapor-diffusion method. The crystals belong to the orthorhombic space group C222(1) with cell dimensions of a = 47.6, b = 106.8 and c = 128.2 A. The crystals diffract to about 2.5 A resolution on a rotating-anode X-ray source
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three-dimensional structure analysis, catalytic triad consisting of serine, histidine and aspartic acid
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vapor diffusion hanging drop method. X-ray crystal structures (at 1.54 and 1.76 A resolution) of HbHNL complexes with two chiral substrates (S)-mandelonitrile and (2S)-2,3-dimethyl-2-hydroxybutyronitrile by soaking and rapid freeze quenching techniques. Only the S-enantiomers of the two substrates are observed in the active site
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acetone-complexed crystals belong to the tetragonal space group P4(1)2(1)2 with unit-cell parameters a = 105.5 A, c = 188.5 A. Chloroacetone-complexed crystals are isomorphous to acetone-complexed crystals, with unit-cell parameters a = 105.9 A and c = 188.4 A
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vapor diffusion hanging drop method, crystal structure of the mutant enzyme S80A refined to an R-factor of 18.0% against diffraction data to 2.1 A resolution, determination of three-dimensional structure of the complex of mutant enzyme S80A with acetone cyanohydrin at 2.2 A resultion, refined to an R-factor of 18.7%
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vapor-diffusion hanging-drop method. The crystal structure of the mutant W128A substrate free form at 2.1 A resolution indicates that the W128A substitution has significantly enlarged the active-site channel entrance. The MeHNL-W128A/4-hydroxybenzaldehyde complex structure at 2.1 A resolution shows the presence of two hydroxybenzaldehyde molecules in a sandwich type arrangement in the active site with an additional hydrogen bridge to the reacting center
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2.4 - 11
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728015
3
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half-life: 47 h
691345
3.5
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enzyme is inactivated very fast. Stability decreases with higher buffer concentrations at pH 3.5 in all three buffer systems (phosphate buffer, glutamate buffer, citrate buffer)
705188
4.5
-
below pH 4.5 enzyme is inactivated within a few min
651011
4.8
-
still stable at this value
653957
5 - 10
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starts to precipitate below pH 5.0, denatures above pH 10.0
652556
5 - 8
-
half-life: above 100 h
691345
5.4
-
most stable at
651012
6
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very stable above pH 6.0
650346
6.5
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very stable above. At pH 6.5 stability increases with increasing buffer concentration in all three systems (phosphate buffer, glutamate buffer, citrate buffer). The enzyme is most stable in phosphate buffer whereas for glutamate buffer the relative increase of half-life with concentration is more pronounced
705188
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0 - 60
-
the enzyme is stable and exhibits more than 96 h half-life time between 0 and 20C, the enzyme shows more than 48 h half-life time at 30C, 64 h half-life time at 37C, 2.7 h half-life time at 50C, and 0.5 h half-life time at 60C
20
-
half-life time: more than 96 h
30
-
half-life: 1755 min in phosphate buffer, 1379 min in glutamate buffer, 2315 min in citrate buffer
37
-
half-life time: 64 h
40 - 50
-
the enzyme shows a half-life time of 411 h at 40C. At 50C a rapid inactivation is observed
40
-
half-life: 690 min in phosphate buffer, 204 min in glutamate buffer, 322 min in citrate buffer
45
-
mutant enzyme and wild-type enzyme G113S are stable for 20 min at pH 5.4
65
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20 min at pH 5.4, mutant enzyme G113S loses about 15% of its initial activity, wild-type enzyme loses about 50% of its initial activity
additional information
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the enzyme is much more thermostable in 1-propyl-3-methylimidazolium tetrahydroborate than in acetonitrile or tetrahydrofuran
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
half-life at pH 5 and 20C is halved at a concentration of 2 mM benzaldehyde and 30 mM HCN, stability decreases drastically with increasing concentrations of benzaldehyde
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half-life increases linearly with enzyme concentration in the aqueous buffer as well as in a two-phase system of buffer and dibutyl ether
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HNL is stabilized by entrapment in aggregates of Escherichia coli cell proteins
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influence of different organic solvents and solvent mixtures on the stability in two-phase systems of aqueous buffer and organic solvent
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the enzyme is stabilized in crude cell extract
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the enzyme shows especially high stability in mixtures of methyl-tert-butylether and hexane, 40:60 and 25:75
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the immobilized enzyme is successfully used for more than 20 repeated batches with no loss of conversion rate or enantioselectivity
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the use of solvent mixtures with methyl-tert-butylether leads to better stability than the use of pure solvents
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ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dibutyl ether
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inactivates within a few min
Ethyl acetate
-
inactivates within a few min
hexane
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inactivates within a few min, the use of solvent mixtures with methyl-tert-butylether leads to better stability than the use of pure solvents the enzyme shows especially high stability in mixtures of methyl-tert-butylether and hexane, 40:60 and 25:75
methyl-tert-butylether
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the use of solvent mixtures with methyl-tert-butylether leads to better stability than the use of pure solvents the enzyme shows especially high stability in mixtures of methyl-tert-butylether and hexane, 40:60 and 25:75
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
unstable below pH 5, fivefold increase in half-life under optimal conditions by Venoruton and monohydroxyethylrutoside, 50% increase in half-life by 5-20 ng/ml rutin and 1.5-6 ng/ml by hyperoside
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651390, 655659
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4C, stable for up to 12 months
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
mutant enzymes Hnl-E79A, Hnl-S80A, Hnl-C81S, and Hnl-H235A and the wild-type protein are purified to homogeneity, and Hnl-H10A is partially purified, by ion exchange chromatography and native polyacrylamide gel electrophoresis
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mutant K236L, recombinant
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preparation of a cross-linked enzyme aggregate (CLEA) from hydroxynitrile lyase from Hevea brasiliensis, precipitated hydroxynitrile lyase, comparison of free, cross-linked enzyme aggregate (CLEA) and immobilized in sol-gel (aqua gel)
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Q-Sepharose column chromatography; recombinant enzyme
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recombinant enzyme
wild-type and W128A mutant partially purified
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloned and produced by heterologous expression in different microbial hosts
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expressed in Escherichia coli
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expressed in Escherichia coli BL21 (DE3)
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expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli BL21(DE3) cells; expression in Escherichia coli
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expressed in Escherichia coli JM109 and BL21 (DE3) cells
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expressed in Pichia pastoris, Leishmania tarentolae, Escherichia coli strain JM109 and in two cell-free translations, including an Escherichia coli lysate (WakoPURE system) and wheat germ translation system; wild type and mutant H103L are expressed in Escherichia coli strain JM109, in Pichia pastoris and Leishmania tarentolae, as well as in an Escherichia coli lysate (WakoPURE system) and wheat germ translation system
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expressed in roots of Manihot esculenta cultivar TMS-60444
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expressed in the yeast strain Pichia pastoris
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expression in Escherichia coli
expression of mutant and wild-type proteins in Saccharomyces cerevisiae
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expression of mutant K236L in Pichia pastoris
-
full-length cDNA functinal expressed in Escherichia coli, Pichia pastoris and Saccharomyces cerevisiae
-
high expression in a multi-auxotrophic mutant of Saccharomyces cerevisiae
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limitations in enzyme supply from natural resources are overcome by production of the enzyme in the microbial host systems Escherichia coli, Sachycharomyces cerevisiae, and Pichia pastoris. Expression of Hnl in the prokaryotic system leads to the formation of inclusion bodies whereas in both yeast hosts high levels of soluble protein are obtained. Highest yields are obtained in a high cell density batch fermentation of a Pichia pastoris transformant that expresses heterologous Hnl to about 50% of the soluble cytosolic protein. At a cell density of 100 g/liter cell dry weight, a volume yield of 22 g/liter of heterologous product is obtained. Attempts to produce the Hnl protein extracellularly with the yeast hosts by applying different leader peptide strategies are not successful. Immunofluorescence microscopy studies indicate that the secretion-directed heterologous Hnl protein accumulates in the plasma membrane forming aggregated clusters of inactive protein
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overexpression in Pichia pastoris
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overexpression of the enzyme in transgenic Manihot esculenta plants under the control of a double 35S CaMV promoter. Enzyme activity increases more than 2fold in leaves 13fold in roots of transgenic plants relative to wild-type plants. Elevated levels of hydroxynitrile lyase levels are correlated with substantially reduced acetone cyanohydrin levels and increased cyanide volatilization in processed or homogenized roots. Unlike acyanogenic cassava, transgenic plants overexpressing the enzyme in root retain herbivore deterrence of cyanogens while providing a safer food product (cyanide toxicity)
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recombinant Pichia pastoris strains are constructed which simultaneously expressed the (S)-oxynitrilase of Manihot esculenta and the arylacetonitrilase of Pseudomonas fluorescens EBC191 each under the control of individual AOX1 promoters in order to obtain a whole cell catalyst for the synthesis of (S)-mandelic acid from benzaldehyde and cyanide production of optically active cyanohydrin compounds
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succesfully overexpressed in Escherichia coli, Saccharomyces cerevisiae and Pichia pastoris
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the enzyme activity and yield at low-temperature cultures (17C) are 850times higher than those obtained at the optimum growth temperature of 37C
the overexpression of HNL driven by the patatin (root-specific) promoter results in a 2-20fold increase in relative mRNA expression in roots when compared with wild type, and a 5-6fold increase in expression when compared with CaMV 35S HNL transgenic lines
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D207A
mutant protein is insoluble
H235A
mutant protein is insoluble
S80A
mutant protein is insoluble
C81A
-
about 20% of wild-type activity
H103L/W128A
-
increased activity with the substrate (2S)-hydroxy[4-(methoxy)cyclohex-3-en-1-yl]ethanenitrile compared to the starting clone W128A
H10A
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mutation results in a 30000 Da protein with increased electrophoretic mobility on native high percentage (16%) polyacrylamide gels. the mutant enzyme displays almost wild-type specific activity in crude extracts, suggesting that His10 is not crucial for activity. However, activity is almost completely lost during purification, supporting the possibility that the H10A exchange has a destabilizing effect and may prevent formation of an active dimer of the enzyme after purification
K236L
-
inactive mutant protein, three-dimensional structure is similar to wild-type enzyme
K236R
-
0.15% of wild-type activity
P207A
-
no expression
T11A
-
2% of wild-type activity
W128A
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higher conversion and better selectivity than wild-type enzyme with the substrate 4-methoxycyclohex-3-ene-1-carbaldehyde; lower conversion and lower selectivity than wild-type enzyme with the substrate 4-methoxycyclohex-3-ene-1-carbaldehyde
W128A/I219V
-
very weak activity in cleavage reaction with (2S)-hydroxy[4-(methoxy)cyclohex-3-en-1-yl]ethanenitrile
W128A/K147R
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very weak activity in cleavage reaction with (2S)-hydroxy[4-(methoxy)cyclohex-3-en-1-yl]ethanenitrile
W128A/P187L
-
no activity in cleavage reaction with (2S)-hydroxy[4-(methoxy)cyclohex-3-en-1-yl]ethanenitrile
W128A/Q215H
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slightly increased activity with the substrate (2S)-hydroxy[4-(methoxy)cyclohex-3-en-1-yl]ethanenitrile compared to the starting clone W128A
D208A
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Km-value for 2-hydroxy-2-methylpropanenitrile increases from 101 mM for wild-type enzyme to over 200 mM for the mutant enzyme
G113S
-
enhanced thermal stability compared to wild-type enzyme, mutant enzyme retains slight higher activity than the wild-type enzyme in an acidic environment, so the mutant enzyme maybe more effective for synthesis of (S)-cyanohydrin than the wild-type enzyme
H103C
-
the mutant displays 9.3fold increase in total specific activity in the cell-free extract compared with the wild type
H103I
-
the mutant displays 8.1fold increase in total specific activity in the cell-free extract compared with the wild type
H103K
-
inactive
H103M
-
the mutant displays 9.07fold increase in total specific activity in the cell-free extract compared with the wild type
H103P
-
inactive
H103Q
-
the mutant displays 4.06fold increase in total specific activity in the cell-free extract compared with the wild type
H103R
-
inactive
H103S
-
the mutant displays 2.9fold increase in total specific activity in the cell-free extract compared with the wild type
H103T
-
the mutant displays 3.4fold increase in total specific activity in the cell-free extract compared with the wild type
H103W
-
inactive
H103Y
-
inactive with (S)-mandelonitrile as substrate
H10A
-
limited decrease in activity
H112A
-
limited decrease in activity
H236A
-
mutant enzyme is unable to catalyze the decomposition of 2-hydroxy-2-methylpropanenitrile
H5A
-
limited decrease in activity
K176P
-
the mutant displays 2.02fold increase in total specific activity in the cell-free extract compared with the wild type
K176P/K199P/K224P
-
the mutant displays 6.97fold increase in total specific activity in the cell-free extract compared with the wild type
K176P/K224P
-
the mutant displays 5.05fold increase in total specific activity in the cell-free extract compared with the wild type
K199P
-
the mutant displays 1.38fold increase in total specific activity in the cell-free extract compared with the wild type
K199P/K224P
-
the mutant displays 4.25fold increase in total specific activity in the cell-free extract compared with the wild type
K224P
-
the mutant displays 2.53fold increase in total specific activity in the cell-free extract compared with the wild type
S80A
-
mutant enzyme is completely inactive in the 2-hydroxy-2-methylpropanenitrile cleaving assay. No differences to wild type MeHNL according to oligomeric structure, molecular weight, and behavior in the standard purification procedure
T11A
-
specific activity is 24fold lower than that of the wild-type enzyme
additional information
C-terminally truncated enzymes exhibit activities of 75, 102, 65 and 48 U/l for enzyme after removing 2, 4, 6, and 8 amino acids, respectively
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
analysis
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cyanide-based high-throughput screening assay is developed. The assay is useful to detect activity and enantioselectivity of hydroxynitrile lyases theoretically towards any cyanohydrin substrate
drug development
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industrial processes
food industry
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root-specific expression of cassava HNL not only increases total root protein levels 3fold approaching the target values for a nutritionally balanced meal but accelerates cyanogenesis during food processing resulting in a safer and more nutritious food product
pharmacology
synthesis