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
4.1.2.47
-
RECOMMENDED NAME
GeneOntology No.
(S)-hydroxynitrile lyase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
an aliphatic (S)-hydroxynitrile = cyanide + an aliphatic aldehyde or ketone
show the reaction diagram
-
-
-
-
an aliphatic (S)-hydroxynitrile = cyanide + an aliphatic aldehyde or ketone
show the reaction diagram
proposed reaction mechanism: in free enzyme, the hydroxyl group of Ser80 is hydrogen-bonded to the imidazole nitrogen of His236, which in turn is stabilized by a hydrogen bond with Asp208. If the enzyme encounters a substrate molecule, the proton of Ser80 is rapidly transferred to an imidazole nitrogen of His236. The resulting oxyanion of Ser80 functions as a strong base in a nucleophilic attack of the substrate hydroxyl proton, leading to a negatively charged oxyanion on the substrate. This oxyanion could be stabilized by an oxygen hole formed by amide nitrogen of the backbone of Ser80 and Gly78. Both residues belong to the consensus motif, which is typical for nucleophils located in a catalytic triad. The stabilized oxyanion could further increase the local positive charge on the alpha-C atom of the cyanohydrin, allowing the cyanide group to leave the molecule
-
an aromatic (S)-hydroxynitrile = cyanide + an aromatic aldehyde
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
condensation
-
-
cyanohydrin formation
-
-
cyanohydrin formation
-
-
Henry reaction
-
-
PATHWAY
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
Cyanoamino acid metabolism
-
Metabolic pathways
-
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].
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
(R)-LuHNL
-
-
(R)-Oxynitrilase
-
-
(S)-cyanohydrin producing hydroxynitrile lyase
P52705
-
(S)-Hb-HNL
-
-
(S)-HbHNL
P52704
-
(S)-Hydroxynitrile lyase
-
-
(S)-Hydroxynitrile lyase
P52704
-
(S)-Hydroxynitrile lyase
P52705
-
(S)-Me-HNL
-
-
(S)-MeHNL
-
-
(S)-Oxynitrilase
-
-
(S)-selective HNL
-
-
(S)-selective HNL
-
-
(S)-selective hydroxynitrile lyase
-
-
(S)-selective hydroxynitrile lyase
-
-
(S)-selective hydroxynitrile lyase
P52705
-
alpha-Hydroxynitrile lyase
-
-
EC 4.1.2.39
-
formerly
EC 4.1.2.39
P52705
formerly
Hb-HNL
P52704
-
Hydroxynitrile lyase
P52704
-
Hydroxynitrile lyase
-
-
Hydroxynitrile lyase
P52705
-
MeHNKL
P52705
-
mut-HNL1
-
-
Oxynitrilase
P52704
-
Oxynitrilase
-
-
S-hydroxynitrile lyase
P52704
-
S-hydroxynitrile lyase
P52705
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(1S)-1-furan-2-yl-2-nitroethanol
furan-2-carbaldehyde + CH3NO2
show the reaction diagram
-
enantiomeric excess: ~90%, yield: 60-70%
-
-
r
(1S)-2-nitro-1-(4-nitrophenyl)ethanol
4-nitrobenzaldehyde + CH3NO2
show the reaction diagram
-
enantiomeric excess: ~90%, yield: 60-70%
-
-
r
(1S)-2-nitro-1-phenylethanol
benzaldehyde + CH3NO2
show the reaction diagram
-
enantiomeric excess: ~90%, yield: 60-70%
-
-
r
(1S,2R)-2-nitro-1-phenyl-propanol
benzaldehyde + C2H5NO2
show the reaction diagram
-
4 diastereomers, enantiomeric excess: 95%, yield: 67%
-
-
r
(2S)-1-nitrooctan-2-ol
heptanal + CH3NO2
show the reaction diagram
-
enantiomeric excess: ~90%, yield: 60-70%
-
-
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
-
-
?
(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
-
-
?
(2S)-2-hydroxy-2-methylpentanenitrile
cyanide + pentan-2-one
show the reaction diagram
P52705
-
-
-
?
(2S)-hydroxy(3-phenoxyphenyl)ethanenitrile
cyanide + 3-phenoxybenzaldehyde
show the reaction diagram
P52704
-
-
-
r
(R)-2-(2-furyl)-2-hydroxyacetonitrile
furan-2-carbaldehyde + HCN
show the reaction diagram
-
enantiomeric excess: > 99%, yield: 90%
-
-
r
(S)-2-nitro-1-phenylethanol
benzaldehyde + nitromethane
show the reaction diagram
-
-
-
-
r
(S)-2-nitro-1-phenylethanol
nitromethane + benzaldehyde + (R)-2-nitro-1-phenylethanol
show the reaction diagram
-
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
-
-
r
(S)-3-phenoxybenzaldehyde cyanohydrin
m-phenoxybenzaldyhyde + HCN
show the reaction diagram
-
enantiomeric excess: > 98.5%, yield: 95.5%, used for insecticide synthesis
-
-
r
(S)-Mandelonitrile
Cyanide + benzaldehyde
show the reaction diagram
-
-
-
-
?
(S)-Mandelonitrile
Cyanide + benzaldehyde
show the reaction diagram
-
-
-
-
r
(S)-Mandelonitrile
Cyanide + benzaldehyde
show the reaction diagram
-
-
-
-
?
(S)-Mandelonitrile
Cyanide + benzaldehyde
show the reaction diagram
-
-
-
-
r
(S)-Mandelonitrile
Cyanide + benzaldehyde
show the reaction diagram
P52704
-
-
-
?
(S)-Mandelonitrile
Cyanide + benzaldehyde
show the reaction diagram
-
-
-
-
?
(S)-Mandelonitrile
Cyanide + benzaldehyde
show the reaction diagram
-
-
-
-
r
(S)-Mandelonitrile
Cyanide + benzaldehyde
show the reaction diagram
-
-
-
-
?
(S)-Mandelonitrile
Cyanide + benzaldehyde
show the reaction diagram
-
-
-
-
r
(S)-Mandelonitrile
Cyanide + benzaldehyde
show the reaction diagram
-
binding mode of the chiral substrate is identical to that observed for the biological substrate 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
-
-
?
(S)-Mandelonitrile
Cyanide + benzaldehyde
show the reaction diagram
-
enzyme kinetics in both directions is studied on a model system with mandelonitrile, benzaldehyde, and HCN using two different methods: initial rate measurements and progress curve analysis. Ordered Uni bi mechanism with the formation of a dead-end complex of enzyme, (S)-mandelonitrile and HCN. HCN is the first product released from the enzyme followed by benzaldehyde while in the synthesis reaction, benzaldehyde is the first substrate bond to the enzyme followed by HCN
-
-
r
(S)-Mandelonitrile
Cyanide + benzaldehyde
show the reaction diagram
-
high selectivity towards the (S)-enentiomer
-
-
?
(S)-Mandelonitrile
Cyanide + benzaldehyde
show the reaction diagram
-
the enzymatic reversible conversion of (S)-mandelonitrile to HCN and benzaldehyde can be adequately described by a three-step, reversible-ordered UniBi reaction scheme
-
-
r
(S)-mandelonitrile
HCN + benzaldehyde
show the reaction diagram
-
-
-
-
?
(S)-mandelonitrile
HCN + benzaldehyde
show the reaction diagram
-
-
-
-
?
(S)-mandelonitrile
HCN + benzaldehyde
show the reaction diagram
-
highly (S)-selective
-
?
(S)-mandelonitrile
benzaldehyde + HCN
show the reaction diagram
-
-
-
-
?
(S)-mandelonitrile
benzaldehyde + HCN
show the reaction diagram
-
enantiomeric excess: > 99%, yield: 90%
-
-
r
(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
-
-
r
2-furaldehyde cyanohydrin
2-furaldehyde + HCN
show the reaction diagram
-
aqua gel, 30 min, conversion ratio: 89%, enantiomeric excess: 94%, free enzyme, 30 min, conversion ratio: 89%,enantiomeric excess: 94%
-
-
r
2-Furylaldehyde + cyanide
Furan-3-yl-hydroxyacetonitrile
show the reaction diagram
-
-
-
-
-
2-hydroxy-2-methylpropanenitrile
acetone + HCN
show the reaction diagram
-
-
-
-
2-hydroxy-2-methylpropanenitrile
acetone + HCN
show the reaction diagram
-
-
-
-
r
2-hydroxy-2-methylpropanenitrile
acetone + HCN
show the reaction diagram
-
-
-
-
-
2-hydroxy-2-methylpropanenitrile
acetone + HCN
show the reaction diagram
-
-
-
-
?
2-hydroxy-2-methylpropanenitrile
acetone + HCN
show the reaction diagram
P52705
acetone cyanohydrin spontaneously decomposes to acetone and cyanide at pH above 5.0 or temperatures above 35 C
-
-
-
2-hydroxy-2-methylpropanenitrile
acetone + HCN
show the reaction diagram
-
release of HCN serves as a defense against herbivores and microbial attack of the plant
-
?
2-hydroxy-2-methylpropanenitrile
HCN + acetone
show the reaction diagram
-
-
-
?
2-hydroxy-2-methylpropanenitrile
HCN + acetone
show the reaction diagram
-
-
-
?
2-hydroxy-2-methylpropanenitrile
HCN + acetone
show the reaction diagram
-
liberation of HCN by degradation of acetone cyanohydrin is considerably faster than the consumption of HCN by the formation of (S)-mandelonitrile
-
?
2-hydroxy-2-methylpropanenitrile
HCN + acetone
show the reaction diagram
-
release of HCN serves as a defense against herbivores and microbial attack of the plant
-
?
2-hydroxy-2-methylpropanenitrile
cyanide + acetone
show the reaction diagram
-
-
-
-
?
2-hydroxy-2-methylpropanenitrile
cyanide + acetone
show the reaction diagram
-
-
-
-
?
2-hydroxy-2-methylpropanenitrile
cyanide + acetone
show the reaction diagram
P52705
-
-
-
?
2-hydroxy-2-methylpropanenitrile
cyanide + acetone
show the reaction diagram
-
-
-
-
?
2-hydroxy-2-methylpropanenitrile
cyanide + acetone
show the reaction diagram
-
i.e. acetone cyanohydrin
-
-
?
2-hydroxy-2-methylpropanenitrile
cyanide + acetone
show the reaction diagram
-
i.e. acetone cyanohydrin, catalyzes the decomposition of the achiral alpha-hydroxynitrile 2-hydroxy-2-methylpropanenitrile into HCN and acetone during cyanogenesis of damaged plants
-
-
?
2-hydroxy-2-methylpropanenitrile
cyanide + acetone
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
cyanide + acetone
show the reaction diagram
-
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
-
-
?
2-hydroxyisobutyronitrile
HCN + acetone
show the reaction diagram
-
-
-
?
2-Methyl-2-hydroxybutyronitrile
Butanone + cyanide
show the reaction diagram
-
-
-
-
2-nitro-1-phenylethanol
?
show the reaction diagram
-
-, very low turnover number
-
-
?
2-Pentanone + cyanide
3-Hydroxyhexanonitrile
show the reaction diagram
-
-
-
-
-
2-phenoxybenzaldehyde cyanohydrins
?
show the reaction diagram
-
2-phenoxybenzaldehyde cyanohydrin is converted with lower activity than (S)-mandelonitrile and cyclohexanone cyanohydrile
-
-
?
2-Thienylaldehyde + cyanide
Hydroxythiophen-3-yl-acetonitrile
show the reaction diagram
-
-
-
-
-
3,3-dimethyl-2-butanone + acetone cyanohydrin
(S)-2-hydroxy-2-methyl-3,3-dimethyl-butyronitrile
show the reaction diagram
-
transcyanation
-
-
?
3-[(1S)-1-hydroxy-2-nitroethyl]phenol
3-hydroxybenzaldehyde + CH3NO2
show the reaction diagram
-
enantiomeric excess: ~90%, yield: 60-70%
-
-
r
4-Methoxybenzaldehyde + cyanide
4-Methoxymandelonitrile
show the reaction diagram
-
-
-
-
-
Acetone cyanhydrin
Cyanide + acetone
show the reaction diagram
-
-
-
?
Acetone cyanhydrin
Cyanide + acetone
show the reaction diagram
-
-
-
?
Acetone cyanhydrin
Cyanide + acetone
show the reaction diagram
-
-
-
-
?
acetone cyanhydrin
HCN + acetone
show the reaction diagram
-
-
-
?
acetone cyanhydrin
HCN + acetone
show the reaction diagram
-
S-specific
-
?
acetone cyanohydrin
cyanide + acetone
show the reaction diagram
-
-
-
-
?
acetone cyanohydrin
hydrocyanic acid + acetone
show the reaction diagram
-
-
-
-
?
Acetophenone + cyanide
3-Hydroxy-3-phenylpropionitrile
show the reaction diagram
-
-
-
-
-
acetyltrimethylsilane + acetone cyanohydrin
(S)-2-trimethylsilyl-2-hydroxyl-propionitrile + acetone
show the reaction diagram
-
transcyanation
-
-
?
Benzaldehyde + cyanide
(S)-Mandelonitrile
show the reaction diagram
-
-
-
-
-
cyanide + (2E)-3-(4-hydroxyphenyl)prop-2-enal
(2S,3E)-2-hydroxy-4-(4-hydroxyphenyl)but-3-enenitrile
show the reaction diagram
-
-
wild-type enzyme: 95% enantiomeric excess, 80% conversion rate
-
?
cyanide + (2E)-but-2-enal
(3E)-2-hydroxypent-3-enenitrile
show the reaction diagram
-
-
86% enantiomeric excess with crude enzyme preparation
-
?
cyanide + (2E)-but-2-enal
(3S,3E)-2-hydroxypent-3-enenitrile
show the reaction diagram
-
-
92% enantiomeric excess
-
?
cyanide + (2E)-hex-2-enal
(2S,3E)-2-hydroxyhept-3-enenitrile
show the reaction diagram
-
-
97% enantiomeric excess
-
?
cyanide + (2E)-hex-2-enal
(2S,3E)-2-hydroxyhept-3-enenitrile
show the reaction diagram
-
-
95% enantiomeric excess with crude enzyme preparation
-
?
cyanide + (2E)-hex-2-enal
(2S,3E)-2-hydroxyhept-3-enenitrile
show the reaction diagram
-
-
wild-type enzyme: 97% enantiomeric excess, 58% conversion rate
-
?
cyanide + (2Z)-hex-2-enal
(2S,3Z)-2-hydroxyhept-3-enenitrile
show the reaction diagram
-
-
80% enantiomeric excess with crude enzyme preparation
-
?
cyanide + (4-hydroxyphenyl)acetaldehyde
(2S)-2-hydroxy-3-(4-hydroxyphenyl)propanenitrile
show the reaction diagram
-
-
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
-
-
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
-
?
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
-
-
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
-
-
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
-
-
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
-
-
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
-
-
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
-
-
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
-
-
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
-
-
67% enantiomeric excess
-
?
cyanide + 2,2-dimethylpropanal
(2S)-2-hydroxy-3,3-dimethylbutanenitrile
show the reaction diagram
-
-
94% 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
-
-
92% enantiomeric excess
-
?
cyanide + 2-chlorobenzaldehyde
(2S)-(2-chlorophenyl)(hydroxy)ethanenitrile
show the reaction diagram
-
-
wild-type enzyme: 98% enantiomeric excess, 96% conversion rate
-
?
cyanide + 2-hydroxybenzaldehyde
(2S)-(2-hydroxyphenyl)(hydroxy)ethanenitrile
show the reaction diagram
-
-
wild-type enzyme: 91% enantiomeric excess, 47% conversion rate
-
?
cyanide + 2-methoxybenzaldehyde
(2S)-2-hydroxy-2-(2-methoxyphenyl)acetonitrile
show the reaction diagram
P52704
-
77% enantiomeric excess
-
?
cyanide + 2-methylpropanal
(2S)-2-hydroxy-3-methylbutanenitrile
show the reaction diagram
-
-
81% enantiomeric excess
-
?
cyanide + 2-methylpropanal
(2S)-2-hydroxy-3-methylbutanenitrile
show the reaction diagram
-
-
95% enantiomeric excess
-
?
cyanide + 3,3-dimethylbutan-2-one
(2S)-2-hydroxy-2,3,3-trimethylbutanenitrile
show the reaction diagram
-
-
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)-2-hydroxy-2-(3-methoxyphenyl)acetonitrile
show the reaction diagram
P52704
-
99% enantiomeric excess
-
?
cyanide + 3-phenoxybenzaldehyde
(2S)-2-hydroxy-2-(3-phenoxyphenyl)acetonitrile
show the reaction diagram
P52704
-
99% enantiomeric excess
-
?
cyanide + 3-phenoxybenzaldehyde
(2S)-hydroxy(3-phenoxyphenyl)acetonitrile
show the reaction diagram
-
-
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-phenylpropanal
(2S)-2-hydroxy-4-phenylbutanenitrile
show the reaction diagram
P52704
-
93% enantiomeric excess
-
?
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
-
-
wild-type enzyme: 99% enantiomeric excess, 79% conversion rate
-
?
cyanide + 4-methoxybenzaldehyde
(2S)-2-hydroxy-2-(4-methoxyphenyl)acetonitrile
show the reaction diagram
P52704
-
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
-
-
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
-
-
-
-
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
P52704
-
i.e. (S)-mandelonitrile, more than 99% enantiomeric excess
-
?
cyanide + benzaldehyde
(S)-mandelonitrile
show the reaction diagram
-
-
-
-
?
cyanide + benzaldehyde
(S)-mandelonitrile
show the reaction diagram
-
-
-
-
r
cyanide + benzaldehyde
(S)-mandelonitrile
show the reaction diagram
-
-
-
-
?
cyanide + benzaldehyde
(S)-mandelonitrile
show the reaction diagram
-
-
-
-
r
cyanide + benzaldehyde
(S)-mandelonitrile
show the reaction diagram
-
-
94% enantiomeric excess
-
?
cyanide + benzaldehyde
(S)-mandelonitrile
show the reaction diagram
-
-
98% enantiomeric excess
-
?
cyanide + benzaldehyde
(S)-mandelonitrile
show the reaction diagram
-
-
wild-type enzyme: 99% enantiomeric excess, 97% conversion rate
-
?
cyanide + benzaldehyde
(S)-mandelonitrile
show the reaction diagram
-
-
low enantioselectivity with 20% ee (S)
-
r
cyanide + benzaldehyde
(S)-mandelonitrile
show the reaction diagram
-
concentrations of benzaldehyde higher than 1 M result in decreased enantioselectivity due to nonenzymatic formation of racemic mandelonitrile due to an excess of benzaldehyde and cyanide
-
-
?
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
-
-
80% enantiomeric excess
-
?
cyanide + cinnamaldehyde
(2S)-2-hydroxy-4-phenyl-(E)-but-3-enenitrile
show the reaction diagram
P52704
-
95% enantiomeric excess
-
?
cyanide + cyclohex-3-ene-1-carbaldehyde
(2S)-2-(cyclohex-3-enyl)-2-hydroxyacetonitrile
show the reaction diagram
P52704
-
99% enantiomeric excess
-
?
cyanide + cyclohexanecarbaldehyde
(2S)-2-cyclohexyl-2-hydroxyacetonitrile
show the reaction diagram
P52704
-
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
-
-
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
P52704
-
99% enantiomeric excess
-
?
cyanide + phenylacetaldehyde
2-hydroxy-3-phenylpropanenitrile
show the reaction diagram
-
-
wild-type enzyme: 98% enantiomeric excess, 99% conversion rate
-
?
cyanide + prop-2-enal
(2S)-2-hydroxybut-3-enenitrile
show the reaction diagram
-
-
47% enantiomeric excess
-
?
cyanide + prop-2-enal
(2S)-2-hydroxybut-3-enenitrile
show the reaction diagram
-
-
94% enantiomeric excess with crude enzyme preparation
-
?
cyanide + prop-2-enal
2-hydroxybut-3-enenitrile
show the reaction diagram
-
-
84% enantiomeric excess
-
?
cyanide + prop-2-enal
(2S)-2-hydroxy-3-methylbutanenitrile
show the reaction diagram
-
-
-
-
?
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
P52705
-
-
-
?
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
-
-
67% enantiomeric excess
?
HCN + 3-phenylpropionaldehyde
(2S)-2-hydroxy-4-phenylbutanenitrile
show the reaction diagram
-
-
89% enantiomeric excess
?
HCN + 4-methoxybenzaldehyde
(4-methyoxyphenyl) (hydroxy)acetonitrile
show the reaction diagram
-
-
-
?
HCN + acrolein
(2S)-2-hydroxybut-3-enenitrile
show the reaction diagram
-
-
59% enantiomeric excess
?
HCN + acrolein
(2S)-2-hydroxybut-3-enenitrile
show the reaction diagram
-
-
92% enantiomeric ecxess
?
HCN + allyloxy-2-hydroxypropionitrile
3-allyloxy-2-hydroxypropionitrile
show the reaction diagram
-
-
-
?
HCN + benzaldehyde
(R)-mandelonitrile
show the reaction diagram
-
-
-
?
HCN + benzaldehyde
(R)-mandelonitrile
show the reaction diagram
-
-
-
?
HCN + benzaldehyde
(S)-mandelonitrile
show the reaction diagram
-
-
99% enantiomeric excess
?
HCN + benzaldehyde
(S)-mandelonitrile
show the reaction diagram
-
-
99% enantiomeric excess
?
HCN + benzaldehyde
(S)-mandelonitrile
show the reaction diagram
-
enzyme encapsulated in sol-gel matrix
-
-
?
HCN + benzaldehyde
(S)-mandelonitrile
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 + 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
-
aqua gel, 2 h, conversion ratio: 92%, enantiomeric excess: 94%, free enzyme, 4 h, conversion ratio: 91%,enantiomeric excess: 94%
-
-
r
Isobutyraldehyde + cyanide
2-Hydroxy-3-methylbutyronitrile
show the reaction diagram
-
-
-
-
-
lactonitrile
?
show the reaction diagram
-
poor substrate
-
-
?
m-phenoxybenzaldehyde cyanohydrin
m-phenoxybenzaldehyde + HCN
show the reaction diagram
-
aqua gel, 72 h, conversion ratio: 92%, enantiomeric excess: 98%, free enzyme, 72 h, conversion ratio: 45%,enantiomeric excess: 82%
-
-
r
mandelonitrile
HCN + benzaldehyde
show the reaction diagram
-
-
-
?
mandelonitrile
HCN + benzaldehyde
show the reaction diagram
-
racemic
-
?
mandelonitrile
HCN + benzaldehyde
show the reaction diagram
-
racemic
-
?
n-Butyraldehyde + cyanide
(S)-2-Hydroxyvaleronitrile
show the reaction diagram
-
-
-
-
nitromethane + benzaldehyde
(S)-2-nitro-1-phenylethanol
show the reaction diagram
-
-
-
-
r
phydroxymandelonitrile
?
show the reaction diagram
-
phydroxymandelonitrile is converted with lower activity than (S)-mandelonitrile and cyclohexanone cyanohydrile
-
-
?
propionaldehyde cyanohydrins
?
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
-
?
mandelonitrile
benzaldehyde + HCN
show the reaction diagram
-
aqua gel, 0.5 h, conversion ratio: 97%, enantiomeric excess: 99%, CLEA, 72 h, conversion ratio: 55%, enantiomeric excess: 67%, free enzyme, 4 h, conversion ratio: 97%,enantiomeric excess: 97%
-
-
r
additional information
?
-
-
acetylferrocene and 1,1'-diacetylferrocene are not transformed with this enzyme
-
-
-
additional information
?
-
-
acetylferrocene and 1,1'-diacetylferrocene are not transformed with this enzyme
-
?
additional information
?
-
-
the enantiomeric excess of the product is optimal at pH 5.4 and at HCN concentration between 200 mM and 400 mM and clearly decreases at concentrations greater than 1.5 M. When the temperature decreases from 25C to -5C, the enantiomeric excess increases from 88% to 95%
-
-
-
additional information
?
-
-
the enantiomeric excess of the product is optimal at pH 5.4 and at HCN concentration between 200 mM and 400 mM and clearly decreases at concentrations greater than 1.5 M. When the temperature decreases from 25C to -5C, the enantiomeric excess increases from 88% to 95%
-
?
additional information
?
-
-
When the temperature decreases from 25C to -5C, the enantiomeric excess increases from 67% to 74%
-
-
-
additional information
?
-
-
When the temperature decreases from 25C to -5C, the enantiomeric excess increases from 67% to 74%
-
?
additional information
?
-
-
(3E)-2-hydroxy-4-phenylbut-3-enenitrile is not sufficently accepted by the enzyme in crude enzyme preparation
-
-
-
additional information
?
-
-
modeling of the complexes of the enzyme with its natural substrate acetone cyanohydrin as well as with the chiral compounds mandelonitrile and 2,3-dimethyl-2-hydroxybutyronitril. Enzymatic mechanism involves catalytic triad Ser80, His235, and Asp207 as a genertal acid/base
-
-
-
additional information
?
-
-
the enzyme catalyzes enantioselective addition of HCN to aromatic, heteroaromatic and aliphatic aldehydes and ketones forming (S)-aldehyde cyanohydrins or (S)-ketone cyanohydrins
-
-
-
additional information
?
-
-
theoretical investigation of the catalytic mechanism
-
-
-
additional information
?
-
-
wild-type enzyme shows very low activity with 4-hydroxymandelonitrile, activity of mutant enzyme W128A is increased 450fold compared to wild-type value (Km-value for mutant enzyme W128A is 0.625 mM)
-
-
-
additional information
?
-
-
the enzyme catalyzes asymmetric cyanohydrin and Henry reactions. (R)-2-nitro-1-phenylethanol is not a substrate or HNL
-
-
-
additional information
?
-
-
the enzyme has low activity in performing Henry reactions
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
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
-
-
-
-
r
2-hydroxy-2-methylpropanenitrile
acetone + HCN
show the reaction diagram
-
-
-
-
-
2-hydroxy-2-methylpropanenitrile
acetone + HCN
show the reaction diagram
-
-
-
-
?
2-hydroxy-2-methylpropanenitrile
acetone + HCN
show the reaction diagram
-
release of HCN serves as a defense against herbivores and microbial attack of the plant
-
?
2-hydroxy-2-methylpropanenitrile
HCN + acetone
show the reaction diagram
-
-
-
?
2-hydroxy-2-methylpropanenitrile
HCN + acetone
show the reaction diagram
-
release of HCN serves as a defense against herbivores and microbial attack of the plant
-
?
2-hydroxy-2-methylpropanenitrile
cyanide + acetone
show the reaction diagram
-
-
-
-
?
2-hydroxy-2-methylpropanenitrile
cyanide + acetone
show the reaction diagram
-
i.e. acetone cyanohydrin, catalyzes the decomposition of the achiral alpha-hydroxynitrile 2-hydroxy-2-methylpropanenitrile into HCN and acetone during cyanogenesis of damaged plants
-
-
?
2-hydroxy-2-methylpropanenitrile
cyanide + acetone
show the reaction diagram
P52705
the liberation of HCN serves as a defense mechanism against herbivores and microbial attack in plants
-
-
?
Acetone cyanhydrin
Cyanide + acetone
show the reaction diagram
-
-
-
?
Acetone cyanhydrin
Cyanide + 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 ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
does not contain a flavin group
-
additional information
-
FAD-independent
-
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
-
benzaldehyde
-
-
benzaldehyde
-
acts as a linear competitive inhibitor against mandelonitrile
benzaldehyde
-
strong competitive inhibitor, the half-life of HNL in 20 mM citrate-phosphate buffer (pH 5.0) in the presence of 10 mM of benzaldehyde is around 50 min compared to 450 min in the absence of benzaldehyde; strong inhibitor
Chlorobutanol
-
-
diethyl dicarbonate
-
3 MM, 93% inhibition
diisopropyl fluorophosphate
-
3 mM, 96% inhibition
diisopropyl fluorophosphate
-
both natural MeHNL as well as recombinant MeHNL are almost completely inhibited by diisopropyl fluorophosphate at 1 mM
formaldehyde
-
-
HCN
-
shows S-linear I-parabolic mixed-type inhibition
hexafluoroacetone
-
-
Isobutyraldehyde
-
-
Mandelonitrile
-
-
p-chloromercurybenzoate
-
-
phenylmethylsulfonyl fluoride
-
-
propionaldehyde
-
-
rhodanide
-
very strong competitive inhibitor
Thiocyanate
-
competitive
trichloracetaldehyde
-
-
methyl ethyl ketone
-
-
additional information
-
leupeptin and pepstatin A do not affect the MeHNL-catalyzed 2-hydroxy-2-methylpropanenitrile cleavage at pH 5.4
-
additional information
-
no inhibition by acetate
-
additional information
-
PMSF, NEM or 4-amidinophenylmethanesulfonylfluoride do not inhibit enzymatic activity
-
additional information
-
(R)-2-nitro-1-phenylethanol and nitromethane have negligible inhibition effects on HNL activity
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
dibutyl ether
-
best solvent for HCN concentrations around 300 mM HCN concentration
additional information
-
cleavage of mandelonitrile proceeds more rapidly in monophasic aqueous media containing 1-propyl-3-methylimidazolium tetrahydroborate than in media containing acetonitrile or tetrahydrofuran
-
additional information
-
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 remained good
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
(S)-Mandelonitrile
-
in 50 mM citrate buffer (pH 5.0), temperature not specified in the publication
1.4
-
(S)-Mandelonitrile
-
in 50 mM citrate buffer (pH 5.0), temperature not specified in the publication
1.86
-
(S)-Mandelonitrile
-
mutant enzyme H103M, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
2.5
5
(S)-Mandelonitrile
-
mutant enzyme H103L, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
4.1
-
(S)-Mandelonitrile
-
citrate buffer (50 mM, pH 4.0), at 25C; pH 7.5
4.5
-
(S)-Mandelonitrile
-
pH 5.2, 23C, mutant enzyme W128A
5.17
-
(S)-Mandelonitrile
-
wild type enzyme, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
5.54
-
(S)-Mandelonitrile
-
mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
6.98
-
(S)-Mandelonitrile
-
mutant enzyme K176P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
30
-
(S)-Mandelonitrile
-
pH 5.2, 23C, wild-type enzyme
67
-
2-hydroxy-2-methylpropanenitrile
-
pH 5.2, 23C, wild-type enzyme
95
-
2-hydroxy-2-methylpropanenitrile
-
mutant enzyme H112A
101
-
2-hydroxy-2-methylpropanenitrile
-
recombinant enzyme
120
-
2-hydroxy-2-methylpropanenitrile
-
mutant enzyme D130A; mutant enzyme D95A; wild-type enzyme
125
-
2-hydroxy-2-methylpropanenitrile
-
mutant enzyme H5A
130
-
2-hydroxy-2-methylpropanenitrile
-
mutant enzyme H10A
150
-
2-hydroxy-2-methylpropanenitrile
-
pH 5.2, 23C, mutant enzyme W128A
0.8
-
2-Methyl-2-hydroxybutyronitrile
-
-
161.5
-
3,3-dimethyl-2-butanone
-
pH 5.4, 40C
0.7
-
acetone cyanohydrin
-
-
4
-
acetone cyanohydrin
-
-
115
-
acetone cyanohydrin
-
pH 5.4
169
-
acetone cyanohydrin
-
mutant enzyme C81A
174
-
acetone cyanohydrin
-
wild-type enzyme
61.5
-
acetyltrimethylsilane
-
pH 5.4, 40C
2
3
benzaldehyde
-
wild type enzyme, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
5.9
-
benzaldehyde
-
citrate buffer (50 mM, pH 4.0), at 25C; pH 7.5
12.3
-
benzaldehyde
-
mutant enzyme H103Y, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
13.4
-
benzaldehyde
-
mutant enzyme H103L, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
21.6
-
benzaldehyde
-
mutant enzyme K176P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
27
-
benzaldehyde
-
mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
27.9
-
benzaldehyde
-
mutant enzyme H103M, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
179
-
cyanide
-
citrate buffer (50 mM, pH 4.0), at 25C; pH 7.5
1.1
-
Mandelonitrile
-
pH 5.4, 25C, mutant enzyme G113S
1.4
-
Mandelonitrile
-
pH 5.4, 25C, wild-type enzyme
additional information
-
additional information
-
-
-
additional information
-
additional information
-
wild-type enzyme shows very low activity with 4-hydroxymandelonitrile, activity of mutant enzyme W128A is increased 450fold compared to wild-type value (Km-value for mutant enzyme W128A is 0.625 mM)
-
additional information
-
additional information
-
a kinetic investigation based on the rate curve method
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1.83
-
(S)-Mandelonitrile
-
in 50 mM citrate buffer (pH 5.0), temperature not specified in the publication
21.5
-
(S)-Mandelonitrile
-
mutant enzyme H103L, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
21.67
-
(S)-Mandelonitrile
-
in 50 mM citrate buffer (pH 5.0), temperature not specified in the publication
23.9
-
(S)-Mandelonitrile
-
mutant enzyme H103M, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
34.5
-
(S)-Mandelonitrile
-
wild type enzyme, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
39.7
-
(S)-Mandelonitrile
-
mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
41.8
-
(S)-Mandelonitrile
-
mutant enzyme K176P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
47.4
-
benzaldehyde
-
mutant enzyme H103Y, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
83.2
-
benzaldehyde
-
mutant enzyme K176P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
90.4
-
benzaldehyde
-
mutant enzyme H103L, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
93.5
-
benzaldehyde
-
mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
96.1
-
benzaldehyde
-
mutant enzyme H103M, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
96.6
-
benzaldehyde
-
wild type enzyme, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1.55
-
(S)-Mandelonitrile
-
in 50 mM citrate buffer (pH 5.0), temperature not specified in the publication
467
6
-
(S)-Mandelonitrile
-
mutant enzyme K176P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
467
6.67
-
(S)-Mandelonitrile
-
wild type enzyme, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
467
7.2
-
(S)-Mandelonitrile
-
mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
467
8.43
-
(S)-Mandelonitrile
-
mutant enzyme H103L, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
467
12.8
-
(S)-Mandelonitrile
-
mutant enzyme H103M, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
467
15.83
-
(S)-Mandelonitrile
-
in 50 mM citrate buffer (pH 5.0), temperature not specified in the publication
467
3.44
-
benzaldehyde
-
mutant enzyme H103M, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
7321
3.46
-
benzaldehyde
-
mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
7321
3.85
-
benzaldehyde
-
mutant enzyme H103Y, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication; mutant enzyme K176P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
7321
4.2
-
benzaldehyde
-
wild type enzyme, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
7321
6.75
-
benzaldehyde
-
mutant enzyme H103L, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
7321
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
47.5
-
-
-
57.2
-
-
mutant enzyme G113S
86
-
-
recombinant enzyme
92
-
-
wild-type enzyme from leaves
119.7
-
-
-
240
-
-
native enzyme
250
-
-
recombinant enzyme
additional information
-
-
expressed in Saccharomyces cerevisiae or Pichia pastoris
additional information
-
P52704
screening assay for hydroxynitrile lyases and its application in high-throughput screening of Escherichia coli mutant libraries, semi-quantitative test where the rate of colour formation corresponds to the particular enzyme activity of the sample
additional information
-
-
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
additional information
-
-
no specific activity is detected with recombinant wild type and mutant enzyme H103L when expressed in an Escherichia coli lysate (WakoPURE system) or as N-terminal His6-tagged enzyme in a RTS 100 wheat germ cell-free translation system
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.8
-
-
conversion of formylferrocene
5
-
-
assay at
5.2
-
-
assay at
5.3
5.7
-
substrate: acetone cyanohydrin
5.4
-
-
the enantiomeric excess is optimal
5.5
-
-
mutant enzyme G113S
5.5
-
-
cleavage of mandelonitrile, cleavage of 2-hydroxy-2-methylpropanenitrile
5.5
-
-
immobilized enzyme
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
-
-
to reduce chemical background reaction, pH-values below 5 are necessary
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
5.8
-
pH 4: about 40% of maximal activity, pH 5.8: optimum
4.5
6.5
-
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
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
23
-
-
assay at
25
-
-
assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
-5
25
-
when the temperature decreases from 25C to -5C, the enantiomeric excess increases from 88% to 95%
-5
25
-
When the temperature decreases from 25C to -5C, the enantiomeric excess increases from 74% to 87%
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.3
-
-
isoelectric focusing
5.3
-
-
isoelectric focusing; isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
6% of the activity of leaves
Manually annotated by BRENDA team
-
low expression
Manually annotated by BRENDA team
-
2% of the activity of leaves
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
29210
-
-
electrospray mass spectrometry
30000
-
P52705
recombinant enzyme, SDS-PAGE
50100
-
-
gel filtration
92000
124000
-
-
92000
-
-
gel filtration
100000
105000
-
gel filtration
102000
-
-
gel filtration
124000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 30000
dimer
-
2 * 28500, SDS-PAGE
dimer
-
2 * 42000
homotetramer
-
4 * 29200, in solution, calculated from amino acid sequence
homotrimer
-
3 * 30000, SDS-PAGE
homotrimer
-
3 * 28500, SDS-PAGE
polymer
-
x * 28000-30000
tetramer
-
4 * 25600, SDS-PAGE
tetramer
-
4 * 29000, SDS-PAGE
tetramer
-
the enzyme forms dimers in solution
trimer
-
3 * 29481, calculation from sequence
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
no glycoprotein
-
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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
-
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
-
enzyme in complex with acetone, hexafluoroacetone, trichloroacetaldehyde or rhodanine, hanging-drop vapour diffusion method, space group C222(1)
-
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
-
hanging drop vapor diffusion. crystal structures of hydroxynitrile lyase: one native and three complexes with acetone, isopropyl alcohol, and thiocyanate
-
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
-
three-dimensional structure analysis, catalytic triad consisting of serine, histidine and aspartic acid
-
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
-
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
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3
-
-
half-life: 47 h
3.5
-
-
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)
4
-
-
half-life: 84 h
4
-
-
decativation below pH 4.0, an unfolding of the enzyme followed by aggregation leading to closely packed enzyme particles at low pH values
4
-
-
the enzyme shows a half-life time of ca. 4 h at pH 5.0
4
-
-
half-life time: 2 h, the deactivation of the enzyme at slightly acidic pH is a result of pronounced structural unfolding; the enzyme shows still 40% of its maximal activity at pH 4.0, while its activity is drastically reduced below pH 4.0
4.5
-
-
below pH 4.5 enzyme is inactivated within a few min
4.8
-
-
still stable at this value
5
10
-
starts to precipitate below pH 5.0, denatures above pH 10.0
5
8
-
half-life: above 100 h
5
-
-
half-life in stirred solution is approximately 3.5 h
5
-
-
unstable below. 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
5.4
-
-
most stable at
6
-
-
very stable above pH 6.0
6.5
-
-
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
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
50
-
-
half-life time: 2.7 h
50
-
-
half-life: 342 min in phosphate buffer, 80 min in glutamate buffer, 115 min in citrate buffer
60
-
-
20 min at pH 5.4, mutant enzyme G113S loses about 10% of its initial activity, wild-type enzyme loses about 20% of its initial activity. More than 50% loss of activity of wild-type enzyme after 50 min, 10% loss of activity of mutant enzyme G113 after 50 min, 75% loss of activity of wild-type enzyme after 120 min, 25% loss of activity of mutant enzyme G113 after 120 min
60
-
-
half-life time: 30 min
60
-
-
half-life: 56 min in phosphate buffer, 42 min in glutamate buffer, 29 min in citrate buffer
65
-
-
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
70
-
-
20 min at pH 5.4, about 80% loss of activity, mutant enzyme G113S and wild-type enzyme
70
-
-
half-life: 10 min in phosphate buffer, 15 min in glutamate buffer, 7 min in citrate buffer
additional information
-
-
the enzyme is much more thermostable in 1-propyl-3-methylimidazolium tetrahydroborate than in acetonitrile or tetrahydrofuran
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
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
-
half-life increases linearly with enzyme concentration in the aqueous buffer as well as in a two-phase system of buffer and dibutyl ether
-
influence of different organic solvents and solvent mixtures on the stability in two-phase systems of aqueous buffer and organic solvent
-
the enzyme shows especially high stability in mixtures of methyl-tert-butylether and hexane, 40:60 and 25:75
-
the use of solvent mixtures with methyl-tert-butylether leads to better stability than the use of pure solvents
-
HNL is stabilized by entrapment in aggregates of Escherichia coli cell proteins
-
the enzyme is stabilized in crude cell extract
-
the immobilized enzyme is successfully used for more than 20 repeated batches with no loss of conversion rate or enantioselectivity
-
ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
dibutyl ether
-
inactivates within a few min
Ethyl acetate
-
inactivates within a few min
hexane
-
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
-
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 ACCESSION NO.
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
-
651390, 655659
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4C, stable for up to 12 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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
-
mutant K236L, recombinant
-
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)
-
recombinant enzyme
-
Q-Sepharose column chromatography; recombinant enzyme
-
wild-type and W128A mutant partially purified
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Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
cloned and produced by heterologous expression in different microbial hosts
-
expressed in Escherichia coli
-
expressed in the yeast strain Pichia pastoris
-
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
-
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
-
expressed in Escherichia coli BL21 (DE3)
-
expressed in Escherichia coli BL21(DE3) cells
P52705
expressed in Escherichia coli BL21(DE3) cells; expression in Escherichia coli
-
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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expression in Escherichia coli
-
high expression in a multi-auxotrophic mutant of Saccharomyces cerevisiae
-
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)
-
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
-
succesfully overexpressed in Escherichia coli, Saccharomyces cerevisiae and Pichia pastoris
-
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
the enzyme activity and yield at low-temperature cultures (17C) are 850times higher than those obtained at the optimum growth temperature of 37C
P52705
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 ACCESSION NO.
COMMENTARY
LITERATURE
C81A
-
about 20% of wild-type activity
C81S
-
3% of wild-type activity
C81S
-
no difference can be observed in the electrophoretic mobilities between the wild-type and mutant protein on native polyacrylamide gels and by isoelectric focusing. Mutation does not change the charge or size of the amino acid side chain, but nevertheless greatly reduces activity
E79A
-
1% of wild-type activity
E79A
-
no difference can be observed in the electrophoretic mobilities between the wild-type and mutant protein on native polyacrylamide gels and by isoelectric focusing. Mutation greatly reduces, but does not abolish activity. The negative charge provided by Glu-79 may be required in the active site, but a direct participation of this residue in enzyme catalysis is not suggested
H103L/W128A
-
increased activity with the substrate (2S)-hydroxy[4-(methoxy)cyclohex-3-en-1-yl]ethanenitrile compared to the starting clone W128A
H10A
-
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
H235A
-
no activity
H235A
-
inactive mutant enzyme, no difference can be observed in the electrophoretic mobilities between the wild-type and mutant protein on native polyacrylamide gels and by isoelectric focusing
K236L
-
inactive mutant protein, three-dimensional structure is similar to wild-type enzyme
K236R
-
0.15% of wild-type activity
P207A
-
no expression
S80A
-
no activity
S80A
-
no difference can be observed in the electrophoretic mobilities between the wild-type and mutant protein on native polyacrylamide gels and by isoelectric focusing
T11A
-
2% of wild-type activity
W128A
-
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
-
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
-
slightly increased activity with the substrate (2S)-hydroxy[4-(methoxy)cyclohex-3-en-1-yl]ethanenitrile compared to the starting clone W128A
C81A
-
specific activity is 93% of the wild-type value
C81A
-
the mutant is also inhibited by acetate
D208A
-
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
H103L
-
highly active and soluble mutant
H103L
-
the mutant displays 11.1fold increase in total specific activity in the cell-free extract compared with the wild type
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
W128A
-
substitution of tryptophan128 by an alanine residue enlarges the entrance channel to the active site of MeHNL and thus facilitates access of sterically demanding substrates to the active site, increased conversion rate towards 3-phenoxybenzaldehyde, octan-3-one and heptan-3-one
W128A
-
activity with the natural substrate 2-hydroxy-2-methylpropanenitrile (acetone cyanohydrin) is 70% of wild-type activity. The specific activities of MeHNL-W128A for the unnatural substrates mandelonitrile and 4-hydroxymandelonitrile are increased 9fold and 450fold, respectively, compared with the wild-type. 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, and thereby explains the observed changes in substrate specificity for bulky substrates
W128A
-
mutant enzyme is (S)-selective as the wild-type enzyme
W128A
-
although the variant W128A shows a higher activity with respect to (S)-3-phenoxy-benzaldehyde cyanohydrin, the enantioselectivity is reduced to 85%, compared to 97% of wild-type enzyme
W128C
-
mutation increases the specific activity towards 4-hydroxymandelonitrile of the various MeHNL mutant compared to the wild-type enzyme
W128C
-
mutant enzyme is (S)-selective as the wild-type enzyme
W128L
-
mutation increases the specific activity towards 4-hydroxymandelonitrile of the various MeHNL mutant compared to the wild-type enzyme
W128L
-
mutant enzyme is (S)-selective as the wild-type enzyme
W128Y
-
mutation increases the specific activity towards 4-hydroxymandelonitrile of the various MeHNL mutant compared to the wild-type enzyme
W128Y
-
mutant enzyme is (S)-selective as the wild-type enzyme
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
agriculture
-
enantiomerically pure cyanohydrins produced by enzyme-catalyzed synthesis are important synthetic intermediates for agrochemicals
drug development
-
industrial processes
pharmacology
-
enantiomerically pure cyanohydrins produced by enzyme-catalyzed synthesis are important synthetic intermediates for pharmaceuticals
synthesis
-
of aliphatic and aromatic cyanohydrins
synthesis
-
the enzyme is a potent biocatalyst for the industrial production of chemicals
synthesis
-
industrial important biocatalyst used in enantiospecific syntheses of alpha-hydroxynitriles from aldehydes and methyl-ketones
synthesis
-
enzyme-catalyzed preparation of optically active cyanohydrins
synthesis
-
biocatalyst for the enantiospecific addition of hydrogen cyanide to aldehydes in organic solvents
synthesis
-
industrially relevant C-C-coupling reaction
synthesis
-
production of chiral ferrocene derivatives as ligands in asymmetric catalysis, to bioelectrochemistry and development of new pharmaceuticals against malaria
synthesis
-
catalyzes the industrially interesting formation of (S)-cyanohydrins from aldehydes or ketones and HCN
synthesis
-
crude cell lysate containing a hydroxynitrile lyase can be used for the enantioselective synthesis of several cyanohydrins in a microchannel. These enzymatic reactions show a high initial reaction rate and enantioselectivity, which in a batchwise process can only be achieved by vigorous stirring
synthesis
-
the cyanohydrin reaction of formylferrocene catalysed by the hydroxynitrile lyase from Hevea brasiliensis provides an access to chiral ferrocene derivatives in high enantiomeric excess. Since cyanohydrins are versatile synthetic intermediates, the possibility for many preparative transformations is opened. This synthetic potential is enlarged even further with the transformation of 1,1'-diformylferrocene leading to biscyanohydrins
agriculture
-
enantiomerically pure cyanohydrins produced by enzyme-catalyzed synthesis are important synthetic intermediates for agrochemicals
analysis
-
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
food industry
-
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
-
enantiomerically pure cyanohydrins produced by enzyme-catalyzed synthesis are important synthetic intermediates for pharmaceuticals
synthesis
-
biocatalyst for the enantiospecific addition of hydrogen cyanide to aldehydes in organic solvents
synthesis
-
recombinant Pichia pastoris strains are constructed which simultaneously express 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 cyanideproduction of optically active cyanohydrin compounds
synthesis
-
synthesis of enantiopure (S)-3-phenoxybenzaldehyde cyanohydrin (a useful intermediate in the pyrethroid synthesis) by applying a high pH two-phase system to reduce nonenzymatic reaction
synthesis
-
(S)-mandelonitrile production on a commercial scale
synthesis
-
the enzyme is a valuable catalysts for the synthesis of cyanohydrins, which are versatile chiral building blocks in the pharmaceutical and agrochemical industries
synthesis
-
synthesis of optically active cyanohydrins that are interesting intermediates for the synthesis of alpha-hydroxy acids, alpha-hydroxy ketones, or beta-ethanolamines, all of which are important building blocks in organic synthesis
synthesis
P52705
hydroxynitrile lyase is a useful enzyme for production of optically active cyanohydrin compounds