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Information on EC 4.1.2.47 - (S)-hydroxynitrile lyase
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4.1.2.47 (S)-hydroxynitrile lyaseIUBMB Comments
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].
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Word Map
- 4.1.2.47
- manihot
- esculenta
- brasiliensis
- hevea
- synthesis
- ketone
- cassava
- hcn
- lyases
- benzaldehyde
-
s-mandelonitrile
-
s-cyanohydrins
- rubber
- athnl
-
enantiomeric
-
hydrocyanic
- crantz
-
r-selective
-
s-enantiomer
- arylacetonitrilase
-
cyanogenesis
-
s-mandelic
- montanum
- pharmacology
- agriculture
- drug development
- analysis
- food industry
The enzyme appears in viruses and cellular organisms
Reaction Schemes
=
+
an aliphatic aldehyde or ketone
Synonyms
(R)-LuHNL, (R)-Oxynitrilase, (S)-cyanohydrin producing hydroxynitrile lyase, (S)-Hb-HNL, (S)-HbHNL, (S)-HNL, (S)-Hydroxynitrile lyase, (S)-Me-HNL, (S)-MeHNL, (S)-Oxynitrilase, 
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
(S)-HbHNL
(S)-Hydroxynitrile lyase
(S)-selective HNL
(S)-selective hydroxynitrile lyase
EC 4.1.2.39
Hb-HNL
HbHNL
HNL
Hydroxynitrile lyase
MeHNL
Oxynitrilase
S-HnL
S-hydroxynitrile lyase
S-selective HnL
HNL
-
-
Hydroxynitrile lyase
-
-
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
an aromatic (S)-hydroxynitrile = cyanide + an aromatic aldehyde
-
-
-
-
an aliphatic (S)-hydroxynitrile = cyanide + an aliphatic aldehyde or ketone
-
-
-
-
an aliphatic (S)-hydroxynitrile = cyanide + an aliphatic aldehyde or ketone
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
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
cyanohydrin formation
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PATHWAY SOURCE
PATHWAYS
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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].
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(1S)-1-furan-2-yl-2-nitroethanol
furan-2-carbaldehyde + CH3NO2
-
enantiomeric excess: ~90%, yield: 60-70%
-
-
r
(1S)-2-nitro-1-(4-nitrophenyl)ethanol
4-nitrobenzaldehyde + CH3NO2
-
enantiomeric excess: ~90%, yield: 60-70%
-
-
r
(1S)-2-nitro-1-phenylethanol
benzaldehyde + CH3NO2
-
enantiomeric excess: ~90%, yield: 60-70%
-
-
r
(1S,2R)-2-nitro-1-phenyl-propanol
benzaldehyde + C2H5NO2
-
4 diastereomers, enantiomeric excess: 95%, yield: 67%
-
-
r
(2S)-1-nitrooctan-2-ol
heptanal + CH3NO2
-
enantiomeric excess: ~90%, yield: 60-70%
-
-
r
(2S)-2,3-dimethyl-2-hydroxybutyronitrile
?
-
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)-hydroxy(3-phenoxyphenyl)ethanenitrile
cyanide + 3-phenoxybenzaldehyde
-
-
-
r
(R)-2-(2-furyl)-2-hydroxyacetonitrile
furan-2-carbaldehyde + HCN
-
enantiomeric excess: > 99%, yield: 90%
-
-
r
(S)-2-nitro-1-phenylethanol
nitromethane + benzaldehyde + (R)-2-nitro-1-phenylethanol
-
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
-
enantiomeric excess: > 98.5%, yield: 95.5%, used for insecticide synthesis
-
-
r
(S)-mandelonitrile
cyanide + benzaldehyde + HCN
-
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
3,3-dimethyl-2-butanone + acetone cyanohydrin
(S)-2-hydroxy-2-methyl-3,3-dimethyl-butyronitrile
-
transcyanation
-
-
?
3-[(1S)-1-hydroxy-2-nitroethyl]phenol
3-hydroxybenzaldehyde + CH3NO2
-
enantiomeric excess: ~90%, yield: 60-70%
-
-
r
acetyltrimethylsilane + acetone cyanohydrin
(S)-2-trimethylsilyl-2-hydroxyl-propionitrile + acetone
-
transcyanation
-
-
?
cyanide + (2E)-3-(4-hydroxyphenyl)prop-2-enal
(2S,3E)-2-hydroxy-4-(4-hydroxyphenyl)but-3-enenitrile
-
-
wild-type enzyme: 95% enantiomeric excess, 80% conversion rate
-
?
cyanide + (2E)-but-2-enal
(3E)-2-hydroxypent-3-enenitrile
-
-
86% enantiomeric excess with crude enzyme preparation
-
?
cyanide + (2E)-but-2-enal
(3S,3E)-2-hydroxypent-3-enenitrile
-
-
92% enantiomeric excess
-
?
cyanide + (2Z)-hex-2-enal
(2S,3Z)-2-hydroxyhept-3-enenitrile
-
-
80% enantiomeric excess with crude enzyme preparation
-
?
cyanide + (4-hydroxyphenyl)acetaldehyde
(2S)-2-hydroxy-3-(4-hydroxyphenyl)propanenitrile
-
-
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
-
-
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
-
-
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
-
-
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
-
-
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
-
-
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
-
-
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
-
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
-
-
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
-
-
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
-
wild-type enzyme: 87% enantiomeric excess, 13% conversion rate
-
-
?
cyanide + 1-phenylpropan-2-one
(2S)-2-hydroxy-2-methyl-3-phenylpropanenitrile
-
wild-type enzyme: 97% enantiomeric excess, 82% conversion rate
-
-
?
cyanide + 2,4-dimethylbenzaldehyde
(2S)-(2,4-dimethylphenyl)(hydroxy)ethanenitrile
-
65% yield, 82% enantiomeric excess
-
?
cyanide + 2-bromobenzaldehyde
(2S)-(2-bromophenyl)(hydroxy)ethanenitrile
-
-
wild-type enzyme: 96% enantiomeric excess, 96% conversion rate
-
?
cyanide + 2-bromobenzaldehyde
(S)-2-bromomandelonitrile
-
the (S)-cyanohydrin is formed with more than 99.5% enantiomeric excess
-
-
r
cyanide + 2-chlorobenzaldehyde
(2S)-(2-chlorophenyl)(hydroxy)acetonitrile
-
-
92% enantiomeric excess
-
?
cyanide + 2-chlorobenzaldehyde
(2S)-(2-chlorophenyl)(hydroxy)ethanenitrile
-
-
wild-type enzyme: 98% enantiomeric excess, 96% conversion rate
-
?
cyanide + 2-chlorobenzaldehyde
(S)-2-chloromandelonitrile
-
the (S)-cyanohydrin is formed with more than 99.5% enantiomeric excess
-
-
r
cyanide + 2-flourobenzaldehyde
(S)-2-fluoromandelonitrile
-
the (S)-cyanohydrin is formed with more than 99.5% enantiomeric excess
-
-
r
cyanide + 2-hydroxybenzaldehyde
(2S)-(2-hydroxyphenyl)(hydroxy)ethanenitrile
-
-
wild-type enzyme: 91% enantiomeric excess, 47% conversion rate
-
?
cyanide + 2-methoxybenzaldehyde
(2S)-2-hydroxy-2-(2-methoxyphenyl)acetonitrile
-
77% enantiomeric excess
-
?
cyanide + 2-methylbenzaldehyde
(2S)-(2-methylphenyl)(hydroxy)ethanenitrile
-
76% yield, 47% enantiomeric excess
-
?
cyanide + 3,3-dimethylbutan-2-one
(2S)-2-hydroxy-2,3,3-trimethylbutanenitrile
-
-
78% enantiomeric excess
-
?
cyanide + 3-(4-hydroxyphenyl)propanal
(2S)-2-hydroxy-4-(4-hydroxyphenyl)butanenitrile
-
-
wild-type enzyme: 67% enantiomeric excess, 90% conversion rate
-
?
cyanide + 3-bromobenzaldehyde
(S)-3-bromomandelonitrile
-
the (S)-cyanohydrin is formed with more than 96% enantiomeric excess
-
-
r
cyanide + 3-chlorobenzaldehyde
(S)-3-chloromandelonitrile
-
the (S)-cyanohydrin is formed with more than 96% enantiomeric excess
-
-
r
cyanide + 3-flourobenzaldehyde
(S)-3-fluoromandelonitrile
-
the (S)-cyanohydrin is formed with more than 99% enantiomeric excess
-
-
r
cyanide + 3-hydroxybenzaldehyde
(2S)-(3-hydroxyphenyl)(hydroxy)ethanenitrile
-
-
wild-type enzyme: 97% enantiomeric excess, 88% conversion rate
-
?
cyanide + 3-methoxybenzaldehyde
(2S)-(3-methoxyphenyl)(hydroxy)ethanenitrile
-
67% yield, 76% enantiomeric excess
-
?
cyanide + 3-methoxybenzaldehyde
(2S)-2-hydroxy-2-(3-methoxyphenyl)acetonitrile
-
99% enantiomeric excess
-
?
cyanide + 3-methylbenzaldehyde
(2S)-(3-methylphenyl)(hydroxy)ethanenitrile
-
76% yield, 76% enantiomeric excess
-
?
cyanide + 3-phenoxybenzaldehyde
(2S)-2-hydroxy-2-(3-phenoxyphenyl)acetonitrile
-
99% enantiomeric excess
-
?
cyanide + 3-phenoxybenzaldehyde
(2S)-hydroxy(3-phenoxyphenyl)acetonitrile
-
-
20% enantiomeric excess
-
?
cyanide + 3-phenoxybenzaldehyde
(S)-3-phenoxybenzaldehyde cyanohydrin
-
reaction in a high-pH two-phase system
97% enantiomeric excess
-
r
cyanide + 3-tetrahydrothiophenone
(S)-3-hydroxytetrahydrothiophene-3-carbonitrile
-
-
-
-
?
cyanide + 4-bromobenzaldehyde
(S)-4-bromomandelonitrile
-
the (S)-cyanohydrin is formed with more than 99.5% enantiomeric excess
-
-
r
cyanide + 4-chlorobenzaldehyde
(S)-4-chloromandelonitrile
-
the (S)-cyanohydrin is formed with 93% enantiomeric excess
-
-
r
cyanide + 4-fluorobenzaldehyde
(S)-4-fluoromandelonitrile
-
the (S)-cyanohydrin is formed with more than 99.5% enantiomeric excess
-
-
r
cyanide + 4-hydroxybenzaldehyde
(2S)-(4-hydroxyphenyl)(hydroxy)ethanenitrile
-
-
wild-type enzyme: 94% enantiomeric excess, 51% conversion rate
-
?
cyanide + 4-methoxybenzaldehyde
(2S)-(4-methoxyphenyl)(hydroxy)ethanenitrile
-
-
wild-type enzyme: 99% enantiomeric excess, 79% conversion rate
-
?
cyanide + 4-methoxybenzaldehyde
(2S)-2-hydroxy-2-(4-methoxyphenyl)acetonitrile
-
95% enantiomeric excess
-
?
cyanide + 4-methoxybenzaldehyde
(2S)-hydroxy(4-methoxyphenyl)acetonitrile
-
-
98% enantiomeric excess
-
?
cyanide + 4-methoxycyclohex-3-ene-1-carbaldehyde
(2S)-hydroxy[4-(methoxy)cyclohex-3-en-1-yl]ethanenitrile
-
-
-
-
r
cyanide + 4-methylbenzaldehyde
(2S)-(4-methylphenyl)(hydroxy)ethanenitrile
-
-
wild-type enzyme: 99% enantiomeric excess, 50% conversion rate
-
?
cyanide + 4-methylpentan-2-one
(2S)-2-hydroxy-2,4-dimethylpentanenitrile
-
-
28% enantiomeric excess
-
?
cyanide + 4-oxocyclohexanecarbaldehyde
(2S)-hydroxy(4-oxocyclohexyl)ethanenitrile
-
-
-
-
r
cyanide + 4-phenoxybenzaldehyde
(2S)-(4-phenoxyphenyl)(hydroxy)ethanenitrile
-
-
wild-type enzyme: 96% enantiomeric excess, 47% conversion rate
-
?
cyanide + 4-phenylbutan-2-one
(2S)-2-hydroxy-2-methyl-4-phenylbutanenitrile
-
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
-
-
-
-
r
cyanide + 6-methylhept-5-en-2-one
(2S)-2-hydroxy-2,6-dimethylhept-5-enenitrile
-
wild-type enzyme: 61% enantiomeric excess, 78% conversion rate
-
-
?
cyanide + benzaldehyde
(2S)-2-hydroxy-2-phenylacetonitrile
-
i.e. (S)-mandelonitrile, more than 99% enantiomeric excess
-
?
cyanide + butan-2-one
(2S)-2-hydroxy-2-methylbutanenitrile
-
-
18% enantiomeric excess
-
?
cyanide + butanal
(2S)-2-hydroxypentanenitrile
-
-
80% enantiomeric excess
-
?
cyanide + cinnamaldehyde
(2S)-2-hydroxy-4-phenyl-(E)-but-3-enenitrile
-
95% enantiomeric excess
-
?
cyanide + cyclohex-3-ene-1-carbaldehyde
(2S)-2-(cyclohex-3-enyl)-2-hydroxyacetonitrile
-
99% enantiomeric excess
-
?
cyanide + cyclohexanecarbaldehyde
(2S)-2-cyclohexyl-2-hydroxyacetonitrile
-
99% enantiomeric excess
-
?
cyanide + cyclohexanecarbaldehyde
(2S)-cyclohexyl(hydroxy)acetonitrile
-
-
92% enantiomeric excess
-
?
cyanide + decanal
(2S)-2-hydroxyundecanenitrile
-
-
wild-type enzyme: 78% enantiomeric excess, 65% conversion rate
-
?
cyanide + dodecanal
(2S)-2-hydroxytridecanenitrile
-
-
wild-type enzyme: 71% enantiomeric excess, 80% conversion rate
-
?
cyanide + ferrocenecarboxaldehyde
(R)-(cyanohydroxymethyl)ferrocene
-
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
-
-
92% enantiomeric excess
-
?
cyanide + heptan-3-one
(2S)-2-ethyl-2-hydroxyhexanenitrile
-
wild-type enzyme: 46% enantiomeric excess, 14% conversion rate
-
-
?
cyanide + hexan-2-one
(2S)-2-hydroxy-2-methylhexanenitrile
-
-
80% enantiomeric excess
-
?
cyanide + nonanal
(2S)-2-hydroxydecanenitrile
-
-
wild-type enzyme: 80% enantiomeric excess, 99% conversion rate
-
?
cyanide + octan-3-one
(2S)-2-ethyl-2-hydroxyheptanenitrile
-
wild-type enzyme: 61% enantiomeric excess, 24% conversion rate
-
-
?
cyanide + octanal
(2S)-2-hydroxynonanenitrile
-
-
wild-type enzyme: 79% enantiomeric excess, 96% conversion rate
-
?
cyanide + pentan-2-one
(2S)-2-hydroxy-2-methylpentanenitrile
-
-
69% enantiomeric excess
-
?
cyanide + pentanal
(2S)-2-hydroxyhexanenitrile
-
-
91% enantiomeric excess
-
?
cyanide + phenylacetaldehyde
(2S)-2-hydroxy-3-phenylpropanenitrile
-
99% enantiomeric excess
-
?
cyanide + phenylacetaldehyde
2-hydroxy-3-phenylpropanenitrile
-
-
wild-type enzyme: 98% enantiomeric excess, 99% conversion rate
-
?
cyanide + prop-2-enal
2-hydroxybut-3-enenitrile
-
-
84% enantiomeric excess
-
?
cyanide + propanal
(2S)-2-hydroxybutanenitrile
-
-
91% enantiomeric excess
-
?
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
-
-
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
-
-
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
-
-
-
-
?
furan-3-carbaldehyde + HCN
(2S)-hydroxy(furan-3-yl)ethanenitrile
-
-
92% enantiomeric excess
-
?
HCN + (benzyloxy)acetaldehyde
3-(benzyloxy)-(2S)-2-hydroxy-propanenitrile + 3-(benzyloxy)-(2R)-2-hydroxy-propanenitrile
-
-
50% 3-(benzyloxy)-(2S)-2-hydroxy-propanenitrile and 50% 3-(benzyloxy)-(2R)-2-hydroxy-propanenitrile
?
HCN + 1,1'-diformylferrocene
(R,R)-1,1-bis(cyanohydroxymethyl)ferrocene
-
-
-
?
HCN + 2,2-dimethylpropanal
2-hydroxy-3,3-dimethylbutyronitrile
-
-
-
?
HCN + 2-allyloxyheptanal
(2R,3RS)-3-allyloxy-2-hydroxyoctanenitrile
-
-
-
?
HCN + 2-allyloxyhexanal
(2R,3RS)-3-allyloxy-2-hydroxyheptanenitrile
-
-
-
?
HCN + 2-allyloxypentanal
(2R,3RS)-3-allyloxy-2-hydroxyhexanenitrile
-
-
-
?
HCN + 2-benzyloxypropanal
3-benzyloxy-2-hydroxybutanenitrile
-
-
-
?
HCN + 2-chlorobenzaldehyde
(2-chlorophenyl)(hydroxy)acetonitrile
-
-
-
?
HCN + 2-methoxymethoxypropanal
3-methoxymethoxy-2-hydroxybutanenitrile
-
-
-
?
HCN + 2-methylallyloxyacetaldehyde
3-(2-methylallyloxy)-2-hydroxypropionitrile
-
-
-
?
HCN + 2-methyldihydrofuran
3-hydroxy-2-methyltetrahydrofuran-3-carbonitrile
-
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
-
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
-
-
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
-
-
84.3% (2S)-2-hydroxy-3-naphthalen-1-yl-propionitrile and 15.6% (2R)-2-hydroxy-3-naphthalen-1-yl-propionitrile
?
HCN + 3-phenoxybenzaldehyde
(2S)-hydroxy(3-phenoxyphenyl)acetonitrile
-
-
-
?
HCN + 3-phenoxypropanal
(2S)-2-hydroxy-4-phenoxybutanenitrile + (2S)-2-hydroxy-4-phenoxybutanenitrile
-
-
95.8% (2S)-2-hydroxy-4-phenoxybutanenitrile and 4.2% (2R)-2-hydroxy-4-phenoxybutanenitrile
?
HCN + 4-methoxybenzaldehyde
(4-methoxyphenyl) (hydroxy)acetonitrile
-
-
-
?
HCN + allyloxy-2-hydroxypropionitrile
3-allyloxy-2-hydroxypropionitrile
-
-
-
?
HCN + benzyloxyacetaldehyde
3-benzyloxy-2-hydroxypropionitrile
-
-
-
?
HCN + cyclohexanecarbaldehyde
(2R)-cyclohexyl(hydroxy)acetonitrile
-
-
-
?
HCN + decanal
(S)-2-hydroxyundecanenitrile
-
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
-
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 + furaldehyde
(2R)-furan-2-yl(hydroxy)ethanenitrile
-
enzyme encapsulated in sol-gel matrix
-
-
?
HCN + hexanal
(S)-2-hydroxyoctanenitrile
-
enzyme encapsulated in sol-gel matrix
-
-
?
HCN + m-phenoxybenzaldehyde
(S)-hydroxy-(3-phenoxy-phenyl)acetonitrile
-
enzyme encapsulated in sol-gel matrix
-
-
?
HCN + methoxymethoxyacetaldehyde
2-hydroxy-3-methoxymethoxypropionitrile
-
-
-
?
HCN + methyl isopropyl ketone
(S)-2-hydroxy-2,3-dimethylbutanenitrile
-
enzyme encapsulated in sol-gel matrix
-
-
?
HCN + phenylacetaldehyde
(2R)-2-hydroxy-3-phenylpropanenitrile
-
-
-
?
HCN + rac-2-methyl-3-phenylpropionaldehyde
(2S,3R)-2-hydroxy-3-methyl-4-phenylbutyronitrile
-
-
wild-type and mutant enzymes Y128Y, W128L, W128C, W128A are (S)-selective
-
?
HCN + rac-2-phenylbutyraldehyde
2-hydroxy-3-phenylpentanenitrile
-
-
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
-
-
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
-
-
wild-type and mutant enzymes Y128Y, W128L, W128C, W128A are (S)-selective
-
?
HCN + tetrahydro-2H-3-pyranone
(3R)-3-hydroxytetrahydro-2H-pyran-3-carbonitrile
-
-
enantiomeric excess at pH 4.75 is 48.3%
?
HCN + tetrahydro-3-furanone
(3R)-3-hydroxytetrahydrofuran-3-carbonitrile
-
-
81% enantiomeric excess
?
HCN + thiophen-2-carbaldehyde
hydroxy(thien-2-yl)acetonitrile
-
-
-
?
HCN + thiophen-3-carbaldehyde
hydroxy(thien-3-yl)acetonitrile
-
-
-
?
HCN + undecanal
(S)-2-hydroxydodecanenitrile
-
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
-
-
?
m-phenoxybenzaldehyde cyanohydrin
m-phenoxybenzaldehyde + cyanide
-
2-phenoxybenzaldehyde cyanohydrin is converted with lower activity than (S)-mandelonitrile and cyclohexanone cyanohydrile
-
-
?
p-hydroxymandelonitrile
?
-
phydroxymandelonitrile is converted with lower activity than (S)-mandelonitrile and cyclohexanone cyanohydrile
-
-
?
propionaldehyde cyanohydrin
propionaldehyde + cyanide
-
poor substrate
-
-
?
thiophene-2-carbaldehyde + HCN
(2S)-hydroxy(thiophen-2-yl)ethanenitrile
-
-
96% enantiomeric excess
-
?
thiophene-3-carbaldehyde + HCN
(2S)-hydroxy(thiophen-3-yl)ethanenitrile
-
-
98% enantiomeric excess
-
?
(2S)-2-hydroxy-2-methylbutanenitrile
cyanide + butan-2-one
the liberation of HCN serves as a defense mechanism against herbivores and microbial attack in plants
-
-
?
(S)-mandelonitrile
benzaldehyde + HCN
-
enantiomeric excess: > 99%, yield: 90%
-
-
r
(S)-Mandelonitrile
Cyanide + benzaldehyde
-
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
-
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
-
the enzymatic reversible conversion of (S)-mandelonitrile to HCN and benzaldehyde can be adequately described by a three-step, reversible-ordered Uni–Bi reaction scheme
-
-
r
(S)-Mandelonitrile
Cyanide + benzaldehyde
-
high selectivity towards the (S)-enentiomer
-
-
?
2-furaldehyde cyanohydrin
2-furaldehyde + HCN
-
aqua gel, 30 min, conversion ratio: 89%, enantiomeric excess: 94%
-
-
r
2-furaldehyde cyanohydrin
2-furaldehyde + HCN
-
free enzyme, 30 min, conversion ratio: 89%,enantiomeric excess: 94%
-
-
r
2-hydroxy-2-methylpropanenitrile
acetone + HCN
-
release of HCN serves as a defense against herbivores and microbial attack of the plant
-
?
2-hydroxy-2-methylpropanenitrile
acetone + HCN
acetone cyanohydrin spontaneously decomposes to acetone and cyanide at pH above 5.0 or temperatures above 35 C
-
-
-
2-hydroxy-2-methylpropanenitrile
cyanide + acetone
-
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-hydroxy-2-methylpropanenitrile
cyanide + acetone
-
i.e. acetone cyanohydrin
-
-
?
2-hydroxy-2-methylpropanenitrile
cyanide + acetone
-
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
the liberation of HCN serves as a defense mechanism against herbivores and microbial attack in plants
-
-
?
2-hydroxy-2-methylpropanenitrile
HCN + acetone
-
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
-
release of HCN serves as a defense against herbivores and microbial attack of the plant
-
?
cyanide + (2E)-hex-2-enal
(2S,3E)-2-hydroxyhept-3-enenitrile
-
-
95% enantiomeric excess with crude enzyme preparation
-
?
cyanide + (2E)-hex-2-enal
(2S,3E)-2-hydroxyhept-3-enenitrile
-
-
97% enantiomeric excess
-
?
cyanide + (2E)-hex-2-enal
(2S,3E)-2-hydroxyhept-3-enenitrile
-
-
wild-type enzyme: 97% enantiomeric excess, 58% conversion rate
-
?
cyanide + 2,2-dimethylpropanal
(2S)-2-hydroxy-3,3-dimethylbutanenitrile
-
-
67% enantiomeric excess
-
?
cyanide + 2,2-dimethylpropanal
(2S)-2-hydroxy-3,3-dimethylbutanenitrile
-
-
94% enantiomeric excess
-
?
cyanide + 2-methylpropanal
(2S)-2-hydroxy-3-methylbutanenitrile
-
-
81% enantiomeric excess
-
?
cyanide + 2-methylpropanal
(2S)-2-hydroxy-3-methylbutanenitrile
-
-
95% enantiomeric excess
-
?
cyanide + 3-phenylpropanal
(2S)-2-hydroxy-4-phenylbutanenitrile
-
93% enantiomeric excess
-
?
cyanide + benzaldehyde
(S)-mandelonitrile
-
68% yield, 54% enantiomeric excess
-
?
cyanide + benzaldehyde
(S)-mandelonitrile
-
-
wild-type enzyme: 99% enantiomeric excess, 97% conversion rate
-
?
cyanide + benzaldehyde
(S)-mandelonitrile
-
-
low enantioselectivity with 20% ee (S)
-
r
cyanide + benzaldehyde
(S)-mandelonitrile
-
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 + benzaldehyde
(S)-mandelonitrile
-
the (S)-cyanohydrin is formed with more than 99.5% enantiomeric excess
-
-
r
cyanide + prop-2-enal
(2S)-2-hydroxybut-3-enenitrile
-
-
94% enantiomeric excess with crude enzyme preparation
-
?
cyanide + prop-2-enal
(2S)-2-hydroxybut-3-enenitrile
-
-
47% enantiomeric excess
-
?
HCN + 3-phenylpropionaldehyde
(2S)-2-hydroxy-4-phenylbutanenitrile
-
-
89% enantiomeric excess
?
HCN + 3-phenylpropionaldehyde
(2S)-2-hydroxy-4-phenylbutanenitrile
-
-
67% enantiomeric excess
?
HCN + acrolein
(2S)-2-hydroxybut-3-enenitrile
-
-
92% enantiomeric ecxess
?
HCN + acrolein
(2S)-2-hydroxybut-3-enenitrile
-
-
59% enantiomeric excess
?
HCN + benzaldehyde
(S)-mandelonitrile
-
-
99% enantiomeric excess
?
HCN + benzaldehyde
(S)-mandelonitrile
-
enzyme encapsulated in sol-gel matrix
-
-
?
HCN + benzaldehyde
(S)-mandelonitrile
-
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
-
aqua gel, 2 h, conversion ratio: 92%, enantiomeric excess: 94%
-
-
r
hexanal cyanohydrin
hexanal + HCN
-
free enzyme, 4 h, conversion ratio: 91%,enantiomeric excess: 94%
-
-
r
m-phenoxybenzaldehyde cyanohydrin
m-phenoxybenzaldehyde + HCN
-
aqua gel, 72 h, conversion ratio: 92%, enantiomeric excess: 98%
-
-
r
m-phenoxybenzaldehyde cyanohydrin
m-phenoxybenzaldehyde + HCN
-
free enzyme, 72 h, conversion ratio: 45%,enantiomeric excess: 82%
-
-
r
mandelonitrile
benzaldehyde + HCN
-
aqua gel, 0.5 h, conversion ratio: 97%, enantiomeric excess: 99%
-
-
r
mandelonitrile
benzaldehyde + HCN
-
CLEA, 72 h, conversion ratio: 55%, enantiomeric excess: 67%
-
-
r
mandelonitrile
benzaldehyde + HCN
-
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 25°C to -5°C, 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 25°C to -5°C, the enantiomeric excess increases from 88% to 95%
-
?
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
?
-
-
theoretical investigation of the catalytic mechanism
-
-
-
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
-
-
-
additional information
?
-
-
on the basis of extensive QM/MM MD and RAMD MD simulations, the catalytic mechanism of the enzyme and its substrate delivery and product (HCN) release are explored. The catalytic reaction approximately follows a two-stage mechanism. The first stage involves two fast processes including the proton abstraction of the substrate through a double-proton transfer and the C-CN bond cleavage, while the second stage concerns HCN formation and is rate-determining
-
-
-
additional information
?
-
-
When the temperature decreases from 25°C to -5°C, the enantiomeric excess increases from 67% to 74%
-
-
-
additional information
?
-
-
When the temperature decreases from 25°C to -5°C, the enantiomeric excess increases from 67% to 74%
-
?
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
?
-
-
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)
-
-
-
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(2S)-2-hydroxy-2-methylbutanenitrile
cyanide + butan-2-one
P52705
the liberation of HCN serves as a defense mechanism against herbivores and microbial attack in plants
-
-
?
2-hydroxy-2-methylpropanenitrile
acetone + HCN
-
release of HCN serves as a defense against herbivores and microbial attack of the plant
-
?
2-hydroxy-2-methylpropanenitrile
cyanide + acetone
-
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
P52705
the liberation of HCN serves as a defense mechanism against herbivores and microbial attack in plants
-
-
?
2-hydroxy-2-methylpropanenitrile
HCN + acetone
-
release of HCN serves as a defense against herbivores and microbial attack of the plant
-
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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
diisopropyl fluorophosphate
-
both natural MeHNL as well as recombinant MeHNL are almost completely inhibited by diisopropyl fluorophosphate at 1 mM
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
-
additional information
-
leupeptin and pepstatin A do not affect the MeHNL-catalyzed 2-hydroxy-2-methylpropanenitrile cleavage at pH 5.4
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
dibutyl ether
-
best solvent for HCN concentrations around 300 mM HCN concentration
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
-
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
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2.6
(S)-2-nitro-1-phenylethanol
-
in 50 mM phosphate buffer pH 6.0, at 22°C; in 50 mM phosphate buffer pH 6.0, at 25°C
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
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
120
2-hydroxy-2-methylpropanenitrile
-
mutant enzyme D130A; mutant enzyme D95A; wild-type enzyme
0.4
2-nitro-1-phenylethanol
-
mutant enzyme F125T/L146M, pH and temperature not specified in the publication
0.7
2-nitro-1-phenylethanol
-
mutant enzyme F125T/Y133F, pH and temperature not specified in the publication
1.4
2-nitro-1-phenylethanol
-
mutant enzyme V106F, pH and temperature not specified in the publication
1.6
2-nitro-1-phenylethanol
-
mutant enzyme V106F/L121Y, pH and temperature not specified in the publication
1.8
2-nitro-1-phenylethanol
-
mutant enzyme V106F/F125T, pH and temperature not specified in the publication
5
2-nitro-1-phenylethanol
-
mutant enzyme C81L, pH and temperature not specified in the publication; mutant enzyme F125T, pH and temperature not specified in the publication; mutant enzyme L121Y/F125T/L146M, pH and temperature not specified in the publication; mutant enzyme L121Y/F125T/Y133F, pH and temperature not specified in the publication; mutant enzyme L121Y/L146M, pH and temperature not specified in the publication; mutant enzyme L121Y, pH and temperature not specified in the publication; mutant enzyme L146M, pH and temperature not specified in the publication; mutant enzyme V106F/L121Y/F125T, pH and temperature not specified in the publication; mutant enzyme V106F/L146M, pH and temperature not specified in the publication
6.1
2-nitro-1-phenylethanol
-
wild type enzyme, pH and temperature not specified in the publication
7.1
2-nitro-1-phenylethanol
-
mutant enzyme Y133F, pH and temperature not specified in the publication
2 - 3
benzaldehyde
-
wild type enzyme, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
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
0.6
mandelonitrile
-
mutant enzyme L152F, pH and temperature not specified in the publication
0.7
mandelonitrile
-
mutant enzyme I209G, pH and temperature not specified in the publication
0.8
mandelonitrile
-
mutant enzyme I209A, pH and temperature not specified in the publication
1.4
mandelonitrile
-
mutant enzyme V106F/L121Y, pH and temperature not specified in the publication
1.5
mandelonitrile
-
mutant enzyme F125T/L146M, pH and temperature not specified in the publication
2
mandelonitrile
-
mutant enzyme F125T, pH and temperature not specified in the publication; mutant enzyme V106F/L121Y/F125T, pH and temperature not specified in the publication
2.1
mandelonitrile
-
mutant enzyme F125T/Y133F, pH and temperature not specified in the publication
2.7
mandelonitrile
-
mutant enzyme L121Y, pH and temperature not specified in the publication
3.1
mandelonitrile
-
mutant enzyme L121Y/F125T, pH and temperature not specified in the publication; mutant enzyme Y133F, pH and temperature not specified in the publication
3.3
mandelonitrile
-
wild type enzyme, pH and temperature not specified in the publication
3.6
mandelonitrile
-
mutant enzyme L121Y/F125T/L146M, pH and temperature not specified in the publication
3.7
mandelonitrile
-
mutant enzyme L146M, pH and temperature not specified in the publication
5.3
mandelonitrile
-
mutant enzyme V106F, pH and temperature not specified in the publication
6.7
mandelonitrile
-
mutant enzyme L121Y/L146M, pH and temperature not specified in the publication
8.6
mandelonitrile
-
mutant enzyme F210I, pH and temperature not specified in the publication; mutant enzyme L148F, pH and temperature not specified in the publication
9.4
mandelonitrile
-
mutant enzyme C81L, pH and temperature not specified in the publication
0.4
rac-2-nitro-1-phenylethanol
-
pH 5.0, temperature not specified in the publication, mutant enzyme F125T/L146M
0.7
rac-2-nitro-1-phenylethanol
-
pH 5.0, temperature not specified in the publication, mutant enzyme F125T/Y133F
1.4
rac-2-nitro-1-phenylethanol
-
pH 5.0, temperature not specified in the publication, mutant enzyme V106F
1.6
rac-2-nitro-1-phenylethanol
-
pH 5.0, temperature not specified in the publication, mutant enzyme V106F/L121Y
1.8
rac-2-nitro-1-phenylethanol
-
pH 5.0, temperature not specified in the publication, mutant enzyme V106F/F125T
6.1
rac-2-nitro-1-phenylethanol
-
pH 5.0, temperature not specified in the publication, wild-type enzyme
7.1
rac-2-nitro-1-phenylethanol
-
pH 5.0, temperature not specified in the publication, mutant enzyme Y133F
0.6
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L152F
0.7
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme I209G
0.8
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme I209A
1.4
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme V106F/L121Y
1.5
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme F125T/L146M; pH 5.0, temperature not specified in the publication, mutant enzyme L121F
2
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme F125T; pH 5.0, temperature not specified in the publication, mutant enzyme V106F/L121Y/F125T
2.1
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme F125T/Y133F
2.7
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121T; pH 5.0, temperature not specified in the publication, mutant enzyme L121Y
2.8
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121A
3.1
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121Y/F125T; pH 5.0, temperature not specified in the publication, mutant enzyme Y133F
3.3
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, wild-type enzyme
3.6
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121Y/F125T/L146M
3.7
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121V; pH 5.0, temperature not specified in the publication, mutant enzyme L146M
4
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121M
4.9
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121I
5.3
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme V106F
5.6
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121W
5.7
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121Q
6.6
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121S
6.7
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121Y/L146M
8
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121H
8.1
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121C
8.6
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme F210I; pH 5.0, temperature not specified in the publication, mutant enzyme L148F
9.4
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme C81L
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
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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
0.1
2-nitro-1-phenylethanol
-
mutant enzyme F125T/L146M, pH and temperature not specified in the publication; mutant enzyme L121Y/L146M, pH and temperature not specified in the publication; mutant enzyme V106F/L121Y, pH and temperature not specified in the publication; mutant enzyme V106F, pH and temperature not specified in the publication
0.12
2-nitro-1-phenylethanol
-
mutant enzyme F125T/Y133F, pH and temperature not specified in the publication
0.15
2-nitro-1-phenylethanol
-
wild type enzyme, pH and temperature not specified in the publication
0.2
2-nitro-1-phenylethanol
-
mutant enzyme C81L, pH and temperature not specified in the publication; mutant enzyme V106F/F125T, pH and temperature not specified in the publication; mutant enzyme V106F/L146M, pH and temperature not specified in the publication
0.3
2-nitro-1-phenylethanol
-
mutant enzyme Y133F, pH and temperature not specified in the publication
0.5
2-nitro-1-phenylethanol
-
mutant enzyme L121Y/F125T/L146M, pH and temperature not specified in the publication; mutant enzyme L121Y/F125T/Y133F, pH and temperature not specified in the publication; mutant enzyme L121Y, pH and temperature not specified in the publication; mutant enzyme L146M, pH and temperature not specified in the publication; mutant enzyme V106F/L121Y/F125T, pH and temperature not specified in the publication
0.6
2-nitro-1-phenylethanol
-
mutant enzyme F125T, pH and 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
3
mandelonitrile
-
mutant enzyme I209G, pH and temperature not specified in the publication
7
mandelonitrile
-
mutant enzyme L152F, pH and temperature not specified in the publication; mutant enzyme V106F, pH and temperature not specified in the publication
17
mandelonitrile
-
mutant enzyme L148F, pH and temperature not specified in the publication
20
mandelonitrile
-
mutant enzyme L146M, pH and temperature not specified in the publication
21
mandelonitrile
-
mutant enzyme F210I, pH and temperature not specified in the publication
22
mandelonitrile
-
mutant enzyme Y133F, pH and temperature not specified in the publication
23
mandelonitrile
-
mutant enzyme I209A, pH and temperature not specified in the publication
25
mandelonitrile
-
wild type enzyme, pH and temperature not specified in the publication
32
mandelonitrile
-
mutant enzyme F125T/L146M, pH and temperature not specified in the publication; mutant enzyme F125T/Y133F, pH and temperature not specified in the publication
33
mandelonitrile
-
mutant enzyme C81L, pH and temperature not specified in the publication
43
mandelonitrile
-
mutant enzyme L121Y/F125T, pH and temperature not specified in the publication
45
mandelonitrile
-
mutant enzyme L121Y/F125T/L146M, pH and temperature not specified in the publication
55
mandelonitrile
-
mutant enzyme V106F/L121Y, pH and temperature not specified in the publication
66
mandelonitrile
-
mutant enzyme F125T, pH and temperature not specified in the publication
82
mandelonitrile
-
mutant enzyme V106F/L121Y/F125T, pH and temperature not specified in the publication
106
mandelonitrile
-
mutant enzyme L121Y, pH and temperature not specified in the publication
0.1
rac-2-nitro-1-phenylethanol
-
pH 5.0, temperature not specified in the publication, mutant enzyme F125T/L146M; pH 5.0, temperature not specified in the publication, mutant enzyme V106F
0.12
rac-2-nitro-1-phenylethanol
-
pH 5.0, temperature not specified in the publication, mutant enzyme F125T/Y133F
0.15
rac-2-nitro-1-phenylethanol
-
pH 5.0, temperature not specified in the publication, wild-type enzyme
0.2
rac-2-nitro-1-phenylethanol
-
pH 5.0, temperature not specified in the publication, mutant enzyme V106F/F125T
0.3
rac-2-nitro-1-phenylethanol
-
pH 5.0, temperature not specified in the publication, mutant enzyme Y133F
0.6
rac-2-nitro-1-phenylethanol
-
pH 5.0, temperature not specified in the publication, mutant enzyme F125T
3
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme I209G; pH 5.0, temperature not specified in the publication, mutant enzyme L121T
5
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121H
6
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121Q
7
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L152F; pH 5.0, temperature not specified in the publication, mutant enzyme V106F
9
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121S
15
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121V; pH 5.0, temperature not specified in the publication, mutant enzyme L121W
17
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L148F
20
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121C; pH 5.0, temperature not specified in the publication, mutant enzyme L146M
21
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme F210I
22
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme Y133F
23
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme I209A; pH 5.0, temperature not specified in the publication, mutant enzyme L121A
25
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, wild-type enzyme
32
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme F125T/L146M; pH 5.0, temperature not specified in the publication, mutant enzyme F125T/Y133F
33
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme C81L; pH 5.0, temperature not specified in the publication, mutant enzyme L121I
43
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121M; pH 5.0, temperature not specified in the publication, mutant enzyme L121Y/F125T
45
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121Y/F125T/L146M
55
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme V106F/L121Y
66
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme F125T
77
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121F
82
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme V106F/L121Y/F125T
106
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121Y
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.55
(S)-mandelonitrile
-
in 50 mM citrate buffer (pH 5.0), temperature not specified in the publication
6
(S)-mandelonitrile
-
mutant enzyme K176P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
6.67
(S)-mandelonitrile
-
wild type enzyme, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
7.2
(S)-mandelonitrile
-
mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
8.43
(S)-mandelonitrile
-
mutant enzyme H103L, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
12.8
(S)-mandelonitrile
-
mutant enzyme H103M, in 100 mM citrate buffer pH above 6.0, temperature not specified in the publication
15.83
(S)-mandelonitrile
-
in 50 mM citrate buffer (pH 5.0), temperature not specified in the publication
3.44
benzaldehyde
-
mutant enzyme H103M, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
3.46
benzaldehyde
-
mutant enzyme K176P/K199P/K224P, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
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
4.2
benzaldehyde
-
wild type enzyme, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
6.75
benzaldehyde
-
mutant enzyme H103L, in 100 mM citrate buffer pH 5.0, temperature not specified in the publication
1
mandelonitrile
-
mutant enzyme L148F, pH and temperature not specified in the publication
1.4
mandelonitrile
-
mutant enzyme V106F, pH and temperature not specified in the publication
2.5
mandelonitrile
-
mutant enzyme F210I, pH and temperature not specified in the publication
3.5
mandelonitrile
-
mutant enzyme C81L, pH and temperature not specified in the publication
4
mandelonitrile
-
mutant enzyme I209G, pH and temperature not specified in the publication
5.4
mandelonitrile
-
mutant enzyme L146M, pH and temperature not specified in the publication
7.1
mandelonitrile
-
mutant enzyme Y133F, pH and temperature not specified in the publication
7.4
mandelonitrile
-
wild type enzyme, pH and temperature not specified in the publication
11
mandelonitrile
-
mutant enzyme L152F, pH and temperature not specified in the publication
12
mandelonitrile
-
mutant enzyme L121Y/F125T/L146M, pH and temperature not specified in the publication
14
mandelonitrile
-
mutant enzyme L121Y/F125T, pH and temperature not specified in the publication
15
mandelonitrile
-
mutant enzyme F125T/Y133F, pH and temperature not specified in the publication
21
mandelonitrile
-
mutant enzyme F125T/L146M, pH and temperature not specified in the publication
29
mandelonitrile
-
mutant enzyme I209A, pH and temperature not specified in the publication
33
mandelonitrile
-
mutant enzyme F125T, pH and temperature not specified in the publication
39
mandelonitrile
-
mutant enzyme L121Y, pH and temperature not specified in the publication; mutant enzyme V106F/L121Y, pH and temperature not specified in the publication
41
mandelonitrile
-
mutant enzyme V106F/L121Y/F125T, pH and temperature not specified in the publication
0.6
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121H
1
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L148F
1.1
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121Q
1.4
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121S; pH 5.0, temperature not specified in the publication, mutant enzyme L121T; pH 5.0, temperature not specified in the publication, mutant enzyme V106F
2.5
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme F210I; pH 5.0, temperature not specified in the publication, mutant enzyme L121C
2.6
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121W
3.5
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme C81L
4
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme I209G; pH 5.0, temperature not specified in the publication, mutant enzyme L121V
5.4
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L146M
6.6
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121I
7.1
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme Y133F
7.4
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, wild-type enzyme
8.3
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121A
11
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121M; pH 5.0, temperature not specified in the publication, mutant enzyme L152F
12
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121Y/F125T/L146M
14
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121Y/F125T
15
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme F125T/Y133F
21
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme F125T/L146M
29
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme I209A
33
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme F125T
39
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121Y; pH 5.0, temperature not specified in the publication, mutant enzyme V106F/L121Y
41
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme V106F/L121Y/F125T
51
rac-mandelonitrile
-
pH 5.0, temperature not specified in the publication, mutant enzyme L121F
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.37
benzaldehyde
-
in 50 mM phosphate buffer pH 6.0, at 22°C; in 50 mM phosphate buffer pH 6.0, at 25°C
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.1
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121P
0.13
-
substrate: rac-2-nitro-1-phenylethanol, pH 5.0, temperature not specified in the publication, mutant enzyme L121Y/L146M; substrate: rac-2-nitro-1-phenylethanol, pH 5.0, temperature not specified in the publication, wild-type enzyme
0.15
-
substrate: rac-2-nitro-1-phenylethanol, pH 5.0, temperature not specified in the publication, mutant enzyme C81L
0.17
-
substrate: rac-2-nitro-1-phenylethanol, pH 5.0, temperature not specified in the publication, mutant enzyme V106F
0.2
-
substrate: rac-2-nitro-1-phenylethanol, pH 5.0, temperature not specified in the publication, mutant enzyme F125T/L146M; substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121E
0.21
-
substrate: rac-2-nitro-1-phenylethanol, pH 5.0, temperature not specified in the publication, mutant enzyme F125T/Y133F; substrate: rac-2-nitro-1-phenylethanol, pH 5.0, temperature not specified in the publication, mutant enzyme V106F/L146M
0.27
-
substrate: rac-2-nitro-1-phenylethanol, pH 5.0, temperature not specified in the publication, mutant enzyme F125T
0.29
-
substrate: rac-2-nitro-1-phenylethanol, pH 5.0, temperature not specified in the publication, mutant enzyme V106F/L121Y/F125T; substrate: rac-2-nitro-1-phenylethanol, pH 5.0, temperature not specified in the publication, mutant enzyme Y133F
0.32
-
substrate: rac-2-nitro-1-phenylethanol, pH 5.0, temperature not specified in the publication, mutant enzyme L146M
0.39
-
substrate: rac-2-nitro-1-phenylethanol, pH 5.0, temperature not specified in the publication, mutant enzyme V106F/F125T
0.56
-
substrate: rac-2-nitro-1-phenylethanol, pH 5.0, temperature not specified in the publication, mutant enzyme L121Y/F125T/Y133F
0.62
-
substrate: rac-2-nitro-1-phenylethanol, pH 5.0, temperature not specified in the publication, mutant enzyme L121Y
0.7
0.71
-
substrate: rac-2-nitro-1-phenylethanol, pH 5.0, temperature not specified in the publication, mutant enzyme L121Y/F125T/L146M
0.9
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme I12A
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
1.4
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121G
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
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121T
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
3.8
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121S
4
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121H
5.1
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme I209G
5.9
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121Q
6.7
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme V106F
7
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L148F
12
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L152F
14
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme F210I; substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121W
16
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121C; substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121V
22
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme C81L
24
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L146M
28
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme Y133F
29.2
-
recombinant mutant enzyme H103L expressed in Escherichia coli JM109 cells, using benzaldehyde as substrate, pH and temperature not specified in the publication
30
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121A
33
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, wild-type enzyme
35
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121I
42
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme I209A
52
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme F125T/Y133F; substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121M
53
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme F125T/L146M
55
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121Y/F125T
64
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121Y/F125T/L146M
86
88
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme V106F/L121Y
92
115
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121Y/L146M
119
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121F
140
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme V106F/L121Y/F125T
149
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121Y
additional information
0.7
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme L121N
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
92
-
substrate: rac-mandelonitrile, pH 5.0, temperature not specified in the publication, mutant enzyme F125T
additional information
-
expressed in Saccharomyces cerevisiae or Pichia pastoris
additional information
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
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5.4
5.5
5.8
7.5
-
the maximal activity of HNL in (S)-2-nitro-1-phenylethanol cleavage is at/above pH 7.5
additional information
5.5
-
cleavage of mandelonitrile, cleavage of 2-hydroxy-2-methylpropanenitrile
additional information
-
to reduce chemical background reaction, pH-values below 5 are necessary
additional information
-
to reduce chemical background reaction, pH-values below 5 are necessary
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25
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TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
-5 - 25
-5 - 25
-
when the temperature decreases from 25°C to -5°C, the enantiomeric excess increases from 88% to 95%
-5 - 25
-
When the temperature decreases from 25°C to -5°C, the enantiomeric excess increases from 74% to 87%
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pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5.3
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
5146, 5152, 5159, 650439, 651011, 653957, 653961, 653962, 654007, 663419, 663421, 671565, 691761, 704762, 705349, 711059, 711749, 714712, 748629
-
-
brenda
-
5150, 5154, 5156, 5158, 652556, 653834, 662210, 662488, 691803, 693103, 695171, 701457, 702797, 703209, 703489, 704384, 704752, 705188, 705349, 706644, 706797, 706800, 747531, 748864
UniProt
brenda
precipitated hydroxynitrile lyase, comparison of free, cross-linked enzyme aggregate (CLEA) and immobilized in sol-gel (aqua gel)
-
-
brenda
-
UniProt
brenda
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
-
hydroxynitrile lyases defend plants from herbivores and microbial attack by releasing cyanide from hydroxynitriles
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UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
Sequence
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PDB
SCOP
CATH
UNIPROT
ORGANISM
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
28500
29200
30000
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
homotetramer
homotrimer
tetramer
homotetramer
-
4 * 29200, in solution, calculated from amino acid sequence
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POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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CRYSTALLIZATION/commentary
ORGANISM
UNIPROT
LITERATURE
sitting drop vapor diffusion method, two crystal structures of Baliospermum montanum hydroxynitrile lyase, apo1 and apo2, are determined at 2.55 and 1.9 A, respectively. Structural comparison between Baliospermum montanum (apo2) and S-hydroxynitrile lyase from Hevea brasiliensis with (S)-mandelonitrile bound to the active site reveal that hydrophobic residues at the entrance region of Baliospermum montanum hydroxynitrile lyase form hydrophobic interactions with the benzene ring of the substrate. The flexible structures of these hydrophobic residues are confirmed by a 15 ns molecular dynamics simulation. This flexibility regulates the size of the active site cavity, enabling binding of various substrates to Baliospermum montanum. The high affinity of Baliospermum montanum hydroxynitrile lyase toward substrates containing a benzene ring is also confirmed by comparing the kinetics of Baliospermum montanum hydroxynitrile lyase and S-hydroxynitrile lyase from Manihot esculenta
-
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
-
sitting drop vapor diffusion method, two crystal structures of Baliospermum montanum hydroxynitrile lyase, apo1 and apo2, are determined at 2.55 and 1.9 A, respectively. Structural comparison between Baliospermum montanum (apo2) and S-hydroxynitrile lyase from Hevea brasiliensis with (S)-mandelonitrile bound to the active site reveal that hydrophobic residues at the entrance region of Baliospermum montanum hydroxynitrile lyase form hydrophobic interactions with the benzene ring of the substrate. The flexible structures of these hydrophobic residues are confirmed by a 15 ns molecular dynamics simulation. This flexibility regulates the size of the active site cavity, enabling binding of various substrates to Baliospermum montanum. The high affinity of Baliospermum montanum hydroxynitrile lyase toward substrates containing a benzene ring is also confirmed by comparing the kinetics of Baliospermum montanum hydroxynitrile lyase and S-hydroxynitrile lyase from Manihot esculenta
-
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%
-
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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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
H103C
-
the mutated enzyme shows low enantioselectivity and specific activity for (S)-mandelonitrile synthesis. (S)-Mandelonitrile enantiomeric excess is 60%
H103C/N156D
-
hydroxynitrile lyase of mutant enzyme H103C/N156D is approximately four times higher than that for mutant enzyme H103C. (S)-Mandelonitrile enantiomeric excess is 32%
H103C/N156G
C81L
-
1.3fold increase in kcat for racemic mandelonitrile; the mutant has high enantioselectivity towards (R)-mandelonitrile with a kcat value 1.3times higher compared to the wild type enzyme
C81S
E79A
F125T
-
2.6fold increase in kcat for racemic mandelonitrile, 4.5fold increase in specificity constants (kcat/Km) for racemic mandelonitrile; the mutant has high enantioselectivity towards (R)-mandelonitrile with a kcat value 2.6times and a kcat/Km value 4.5times higher compared to the wild type enzyme
F125T/L146M
-
the mutant shows higher specific activity towards racemic mandelonitrile compared to the wild type enzyme
F125T/Y133F
-
the mutant shows higher specific activity towards racemic mandelonitrile compared to the wild type enzyme
F210I
-
mutant yields mostly insoluble protein, which suggests that the substitution hinders protein folding; the mutant shows lower specific activity towards racemic mandelonitrile compared to the wild type enzyme
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
I12A
-
mutant yields mostly insoluble protein, which suggests that the substitution hinders protein folding
I209A
-
3.9fold increase in specificity constants (kcat/Km) for racemic mandelonitrile; the mutant has higher enantioselectivity towards (R)-mandelonitrile with a kcat/Km value 3.9times higher compared to the wild type enzyme
I209G
-
mutant yields mostly insoluble protein, which suggests that the substitution hinders protein folding, moderate substrate inhibition by rac-mandelonitrile; the mutant shows lower specific activity towards racemic mandelonitrile compared to the wild type enzyme
K236L
-
inactive mutant protein, three-dimensional structure is similar to wild-type enzyme
L121F
-
6.9fold increase in specificity constants (kcat/Km) for racemic mandelonitrile
L121Y
-
5.3fold increase in specificity constants (kcat/Km) for racemic mandelonitrile, 4.2fold increase in kcat for racemic mandelonitrile, the mutant enzyme shows the same high (S)-enantioselectivity (98% ee) as wild-type enzyme. The mutant catalyzes the cleavage of 2-nitro-1-phenylethanol 4.8 times more efficiently than the wild type enzyme, this is similar to the 4.5-fold increase seen for mandelonitrile cleavage; the mutant has high enantioselectivity towards (S)-mandelonitrile with a kcat value 4.2times higher and a kcat/Km value 5.3times higher compared to the wild type enzyme
L121Y/F125T
-
1.7fold increase in kcat for racemic mandelonitrile, 1.9fold increase in specificity constants (kcat/Km) for racemic mandelonitrile; the mutant has high enantioselectivity towards (R)-mandelonitrile with a kcat value 4.2times higher and a kcat/Km value 1.7times higher compared to the wild type enzyme; the mutant shows higher specific activity towards racemic mandelonitrile compared to the wild type enzyme
L121Y/F125T/L146M
-
5.5fold increase in activity over the wild type enzyme; the mutant shows higher specific activity towards racemic mandelonitrile compared to the wild type enzyme
L121Y/L146M
-
the mutant shows higher specific activity towards racemic mandelonitrile compared to the wild type enzyme
L146M
-
the mutant shows lower specific activity towards racemic mandelonitrile compared to the wild type enzyme
L148F
-
the mutant shows lower specific activity towards racemic mandelonitrile compared to the wild type enzyme
L152F
-
1.5fold increase in specificity constants (kcat/Km) for racemic mandelonitrile; the mutant has higher enantioselectivity towards (R)-mandelonitrile with a kcat/Km value 1.5times higher compared to the wild type enzyme
S80A
V106F
-
the mutant shows lower specific activity towards racemic mandelonitrile compared to the wild type enzyme
V106F/L121Y
-
the mutant shows higher specific activity towards racemic mandelonitrile compared to the wild type enzyme
V106F/L121Y/F125T
-
3.3fold increase in kcat for racemic mandelonitrile, 5.5fold increase in specificity constants (kcat/Km) for racemic mandelonitrile; the mutant has high enantioselectivity towards (R)-mandelonitrile with a kcat value 3.3times higher compared to the wild type enzyme
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