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Information on EC 4.1.2.47 - (S)-hydroxynitrile lyase and Organism(s) Manihot esculenta and UniProt Accession P52705
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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
- esculenta
- manihot
- brasiliensis
- hevea
- synthesis
- cassava
- ketone
- hcn
- lyases
- benzaldehyde
-
s-mandelonitrile
- rubber
- athnl
- crantz
-
hydrocyanic
-
s-cyanohydrins
-
r-selective
- arylacetonitrilase
-
s-enantiomer
-
cyanogenesis
- montanum
-
s-mandelic
- pharmacology
- agriculture
- analysis
- drug development
- food industry
The taxonomic range for the selected organisms is: Manihot esculenta
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
=
+
an aliphatic aldehyde or ketone
Synonyms
mehnl, hbhnl, (s)-hydroxynitrile lyase, alpha-hydroxynitrile lyase, s-hnl, luhnl, s-selective hydroxynitrile lyase, hb-hnl, (s)-oxynitrilase, s-hydroxynitrile lyase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
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
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
PATHWAY SOURCE
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].
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
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
(3S,3E)-2-hydroxypent-3-enenitrile
-
92% enantiomeric excess
-
?
cyanide + (4-hydroxyphenyl)acetaldehyde
(2S)-2-hydroxy-3-(4-hydroxyphenyl)propanenitrile
-
wild-type enzyme: 96% enantiomeric excess, 88% conversion rate
-
?
cyanide + 1,3-benzodioxole-5-carbaldehyde
(2S)-1,3-benzodioxol-5-yl(hydroxy)acetonitrile
-
86% enantiomeric excess
-
?
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 + 2,2-dimethylpropanal
(2S)-2-hydroxy-3,3-dimethylbutanenitrile
-
94% 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-methylpropanal
(2S)-2-hydroxy-3-methylbutanenitrile
-
95% 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-phenoxybenzaldehyde
(S)-3-phenoxybenzaldehyde cyanohydrin
reaction in a high-pH two-phase system
97% enantiomeric excess
-
r
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)-hydroxy(4-methoxyphenyl)acetonitrile
-
98% enantiomeric excess
-
?
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-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 + 6-methylhept-5-en-2-one
(2S)-2-hydroxy-2,6-dimethylhept-5-enenitrile
wild-type enzyme: 61% enantiomeric excess, 78% conversion rate
-
-
?
cyanide + butan-2-one
(2S)-2-hydroxy-2-methylbutanenitrile
-
18% 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 + 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
2-hydroxy-3-phenylpropanenitrile
-
wild-type enzyme: 98% enantiomeric excess, 99% conversion rate
-
?
cyanide + prop-2-enal
(2S)-2-hydroxybut-3-enenitrile
-
47% enantiomeric excess
-
?
cyanide + propanal
(2S)-2-hydroxybutanenitrile
-
91% enantiomeric excess
-
?
furan-3-carbaldehyde + HCN
(2S)-hydroxy(furan-3-yl)ethanenitrile
-
92% enantiomeric excess
-
?
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
-
?
3,3-dimethyl-2-butanone + acetone cyanohydrin
(S)-2-hydroxy-2-methyl-3,3-dimethyl-butyronitrile
-
transcyanation
-
-
?
acetyltrimethylsilane + acetone cyanohydrin
(S)-2-trimethylsilyl-2-hydroxyl-propionitrile + acetone
-
transcyanation
-
-
?
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 + 3-phenylpropionaldehyde
(2S)-2-hydroxy-4-phenylbutanenitrile
-
-
67% enantiomeric excess
?
HCN + 4-methoxybenzaldehyde
(4-methoxyphenyl) (hydroxy)acetonitrile
-
-
-
?
HCN + acrolein
(2S)-2-hydroxybut-3-enenitrile
-
-
59% enantiomeric excess
?
HCN + allyloxy-2-hydroxypropionitrile
3-allyloxy-2-hydroxypropionitrile
-
-
-
?
HCN + benzyloxyacetaldehyde
3-benzyloxy-2-hydroxypropionitrile
-
-
-
?
HCN + methoxymethoxyacetaldehyde
2-hydroxy-3-methoxymethoxypropionitrile
-
-
-
?
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 + thiophen-2-carbaldehyde
hydroxy(thien-2-yl)acetonitrile
-
-
-
?
HCN + thiophen-3-carbaldehyde
hydroxy(thien-3-yl)acetonitrile
-
-
-
?
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
-
-
?
(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
cyanide + benzaldehyde
high selectivity towards the (S)-enentiomer
-
-
?
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
-
-
?
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
-
-
?
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 + benzaldehyde
(S)-mandelonitrile
-
98% enantiomeric excess
-
?
cyanide + benzaldehyde
(S)-mandelonitrile
-
wild-type enzyme: 99% enantiomeric excess, 97% conversion rate
-
?
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 + benzaldehyde
(S)-mandelonitrile
-
-
low enantioselectivity with 20% ee (S)
-
r
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)
-
-
?
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
?
-
-
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%
-
?
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
the liberation of HCN serves as a defense mechanism against herbivores and microbial attack in plants
-
-
?
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
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
diisopropyl fluorophosphate
both natural MeHNL as well as recombinant MeHNL are almost completely inhibited by diisopropyl fluorophosphate at 1 mM
additional information
leupeptin and pepstatin A do not affect the MeHNL-catalyzed 2-hydroxy-2-methylpropanenitrile cleavage at pH 5.4
-
additional information
-
leupeptin and pepstatin A do not affect the MeHNL-catalyzed 2-hydroxy-2-methylpropanenitrile cleavage at pH 5.4
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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
150
2-hydroxy-2-methylpropanenitrile
pH 5.2, 23°C, mutant enzyme W128A
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
1.4
(S)-mandelonitrile
-
in 50 mM citrate buffer (pH 5.0), temperature not specified in the publication
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
-
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)
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
21.67
(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
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]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
15.83
(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
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
3.85
benzaldehyde
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
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
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
additional information
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
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5.5
5.5
5.5
cleavage of mandelonitrile, cleavage of 2-hydroxy-2-methylpropanenitrile
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
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
-5 - 25
-
When the temperature decreases from 25°C to -5°C, the enantiomeric excess increases from 74% to 87%
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). HNL_MANES
258
0
29372
Swiss-Prot
other Location (Reliability: 5)
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
28500
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homotetramer
4 * 29200, in solution, calculated from amino acid sequence
tetramer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
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. 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
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%
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D208A
Km-value for 2-hydroxy-2-methylpropanenitrile increases from 101 mM for wild-type enzyme to over 200 mM for the mutant 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
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
H103Q
the mutant displays 4.06fold increase in total specific activity in the cell-free extract compared with the wild type
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
H236A
mutant enzyme is unable to catalyze the decomposition of 2-hydroxy-2-methylpropanenitrile
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
W128A
W128C
mutation increases the specific activity towards 4-hydroxymandelonitrile of the various MeHNL mutant compared to the wild-type enzyme
W128L
mutation increases the specific activity towards 4-hydroxymandelonitrile of the various MeHNL mutant compared to the wild-type enzyme
W128Y
mutation increases the specific activity towards 4-hydroxymandelonitrile of the various MeHNL mutant compared to the wild-type enzyme
C81A
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
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
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
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
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4
the enzyme shows still 40% of its maximal activity at pH 4.0, while its activity is drastically reduced below pH 4.0
704762
4
half-life time: 2 h, the deactivation of the enzyme at slightly acidic pH is a result of pronounced structural unfolding
704762
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0 - 60
the enzyme is stable and exhibits more than 96 h half-life time between 0 and 20°C, the enzyme shows more than 48 h half-life time at 30°C, 64 h half-life time at 37°C, 2.7 h half-life time at 50°C, and 0.5 h half-life time at 60°C
45
-
mutant enzyme and wild-type enzyme G113S are stable for 20 min at pH 5.4
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
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
additional information
-
the enzyme is much more thermostable in 1-propyl-3-methylimidazolium tetrahydroborate than in acetonitrile or tetrahydrofuran
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
the immobilized enzyme is successfully used for more than 20 repeated batches with no loss of conversion rate or enantioselectivity
HNL is stabilized by entrapment in aggregates of Escherichia coli cell proteins
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
high expression in a multi-auxotrophic mutant of Saccharomyces cerevisiae
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
-
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
-
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
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the enzyme activity and yield at low-temperature cultures (17°C) are 850times higher than those obtained at the optimum growth temperature of 37°C
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
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
synthesis
agriculture
-
enantiomerically pure cyanohydrins produced by enzyme-catalyzed synthesis are important synthetic intermediates for agrochemicals
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
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
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
the enzyme is a valuable catalysts for the synthesis of cyanohydrins, which are versatile chiral building blocks in the pharmaceutical and agrochemical industries
synthesis
hydroxynitrile lyase is a useful enzyme for production of optically active cyanohydrin compounds
synthesis
synthesis of pure 2-hydroxycarboxylic acids as valuable synthetic building blocks. Development of bienzymatic cascades to convert aldehydes, via hydrocyanation and subsequent hydration or hydrolysis, into the corresponding (S)-2-hydroxycarboxylic amides and acids. The biocatalysts comprise an (S)-specific hydroxynitrile lyase combined with a nonselective nitrile hydratase or nitrilase. The key to success is preventing racemisation of the intermediate (S)-2-hydroxynitrile while adequately protecting the nitrile-converting enzyme, either in a cross-linked enzyme aggregate (CLEA) or in resting cells.Two biocatalyst systems for the synthesis of (S)-mandelic acid are developed: a combined cross-linked enzyme aggregate (combi-CLEA) of an (S)-hydroxynitrile lyase and a nitrilase, as well as a whole-cell Escherichia coli biocatalyst expressing both enzymes. The nitrilase formed large amounts of mandelicamide, which was remedied by including an amidase in the combi-CLEA as well as by using nitrilasevariants obtained by directed mutagenesis in the whole-cell biocatalyst. Excellent results with more than 95% conversion of benzaldehyde into (S)-mandelic acid with near-quantitative enantiomeric purity are obtained with both biocatalyst systems. Directed mutagenesis of the nitrilase provides an amide-selective whole-cell biocatalyst, which produces (S)-mandelic amide in near-stoichiometric yields. (S)-2-hydroxylalkanoic carboxamides are synthesised in the presence of CLEAs of hydroxynitrile lyase and nitrile hydratase
synthesis
use of biocatalytic active static emulsions (BASE) for the hydroxynitrile lyase-catalyzed synthesis of enantiopure cyanohydrins. With this technique a full suppression of the undesired racemic non-enzymatic side reaction is facilitated, even at unusually high pH within the aqueous phase. BASE is an inclusion immobilization. It consists of a hydrophobic matrix, typically polydimethylsiloxane (PDMS), with dispersed domains of an aqueous phase
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
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
White, W.L.B.; Ariaz-Garzon, D.I.; McMahon, J.M.; Sayre, R.T.
Cyanogenesis in Cassava. The role of hydroxynitrile lyase in root cyanide production
Plant Physiol.
116
1219-1225
1998
Manihot esculenta (P52705)
Lauble, H.; Decanniere, K.; Wajant, H.; Förster, S.; Effenberger, F.
Crystallization and preliminary X-ray diffraction analysis of hydroxynitrile lyase from cassava (manihot esculenta)
Acta Crystallogr. Sect. D
55
904-906
1999
Manihot esculenta
-
Foerster, S.; Roos, J.; Effenberger, F.; Wajant, H.; Sprauer, A.
The first recombinent hydroxynitrile lyase in the synthesis of (S)-cyanohydrins
Angew. Chem. Int. Ed. Engl.
35
437-439
1996
Manihot esculenta (P52705)
-
Hughes, J.; Lakey, J.H.; Hughes, M.A.
Production and characterization of a plant alpha-hydroxynitrile lyase in Escherichia coli
Biotechnol. Bioeng.
53
332-338
1997
Manihot esculenta
Chueskul, S.; Chulavatnatol, M.
Properties of alpha-hydroxynitrile lyase from the petiole of cassava (Manihot esculenta Crantz)
Arch. Biochem. Biophys.
334
401-405
1996
Manihot esculenta
Wajant, H.; Foerster, S.; Spauer, A.; Effenberger, F.; Pfizenmaier, K.
Enantioselective synthesis of aliphatic (S)-cyanohydrins in organic solvents using hydroxynitrile lyase from Manihot esculenta
Ann. N. Y. Acad. Sci.
799
771-776
1996
Manihot esculenta
Hughes, J.; Carvalho, J.P.de C.; Hughes, M.A.
Purification, characterization, and cloning of alpha-hydroxynitrile lyase from cassava (Manihot esculenta Crantz)
Arch. Biochem. Biophys.
311
496-502
1994
Manihot esculenta (P52705), Manihot esculenta
Lauble, H.; Förster, S.; Miehlich, B.; Wajant, H.; Effenberger, F.
Structure of hydroxynitrile lyase from Manihot esculenta in complex with substrates acetone and chloroacetone: implications for the mechanism of cyanogenesis
Acta Crystallogr. Sect. D
57
194-200
2001
Manihot esculenta (P52705)
Yan, G.; Cheng, S.; Zhao, G.; Wu, S.; Liu, Y.; Sun, W.
A single residual replacement improves the folding and stability of recombinant cassava hydroxynitrile lyase in E. coli
Biotechnol. Lett.
25
1041-1047
2003
Manihot esculenta
Bühler, H.; Effenberger, F.; Förster, S.; Roos, J.; Wajant, H.
Substrate specificity of mutants of the hydroxynitrile lyase from Manihot esculenta
ChemBioChem
4
211-216
2003
Manihot esculenta (P52705), Manihot esculenta
Effenberger, F.; Foerster, S.; Wajant, H.
Hydroxynitrile lyases in stereoselective catalysis
Curr. Opin. Biotechnol.
11
532-539
2000
Linum usitatissimum, Manihot esculenta
Costes, D.; Wehtje, E.; Adlercreutz, P.
Hydroxynitrile lyase-catalyzed synthesis of cyanohydrins in organic solvents. Parameters influencing activity and enantiospecificity
Enzyme Microb. Technol.
25
384-391
1999
Hevea brasiliensis, Manihot esculenta
-
Lauble, H.; Miehlich, B.; Förster, S.; Wajant, H.; Effenberger, F.
Mechanistic aspects of cyanogenesis from active-site mutant Ser80Ala of hydroxynitrile lyase from manihot esculenta in complex with acetone cyanohydrin
Protein Sci.
10
1015-1022
2001
Manihot esculenta
Lauble, H.; Miehlich, B.; Förster, S.; Kobler, C.; Wajant, H.; Effenberger, F.
Structure determinants of substrate specificity of hydroxynitrile lyase from Manihot esculenta
Protein Sci.
11
65-71
2002
Manihot esculenta (P52705), Manihot esculenta
Roos, J.; Effenberger, F.
Hydroxynitrile lyase catalyzed enantioselective HCN addition to O-protected alpha-hydroxyaldehydes
Tetrahedron Asymmetry
10
2817-2828
1999
Manihot esculenta
-
Griengl, H.; Schwab, H.; Fechter, M.
The synthesis of chiral cyanohydrins by oxynitrilases
Trends Biotechnol.
18
252-256
2000
Hevea brasiliensis, Manihot esculenta
Xu, R.; Zong, M.H.; Liu, Y.Y.; He, J.; Zhang, Y.Y.; Lou, W.Y.
Enzymatic enantioselective transcyanation of silicon-containing aliphatic ketone with (S)-hydroxynitrile lyase from Manihot esculenta
Appl. Microbiol. Biotechnol.
66
27-33
2004
Manihot esculenta
Lou, W.Y.; Xu, R.; Zong, M.H.
Hydroxynitrile lyase catalysis in ionic liquid-containing systems
Biotechnol. Lett.
27
1387-1390
2005
Manihot esculenta
Bühler, H.; Miehlich, B.; Effenberger, F.
Inversion of stereoselectivity by applying mutants of the hydroxynitrile lyase from Manihot esculenta
ChemBioChem
6
711-717
2005
Manihot esculenta
Siritunga, D.; Arias-Garzon, D.; White, W.; Sayre, R.T.
Over-expression of hydroxynitrile lyase in transgenic cassava roots accelerates cyanogenesis and food detoxification
Plant Biotechnol. J.
2
37-43
2004
Manihot esculenta
Rustler, S.; Motejadded, H.; Altenbuchner, J.; Stolz, A.
Simultaneous expression of an arylacetonitrilase from Pseudomonas fluorescens and a (S)-oxynitrilase from Manihot esculenta in Pichia pastoris for the synthesis of (S)-mandelic acid
Appl. Microbiol. Biotechnol.
80
87-97
2008
Manihot esculenta
von Langermann, J.; Guterl, J.K.; Pohl, M.; Wajant, H.; Kragl, U.
Hydroxynitrile lyase catalyzed cyanohydrin synthesis at high pH-values
Bioprocess Biosyst. Eng.
31
155-161
2008
Manihot esculenta (P52705), Manihot esculenta
Semba, H.; Dobashi, Y.; Matsui, T.
Expression of hydroxynitrile lyase from Manihot esculenta in yeast and its application in (S)-mandelonitrile production using an immobilized enzyme reactor
Biosci. Biotechnol. Biochem.
72
1457-1463
2008
Manihot esculenta (P52705), Manihot esculenta
Van Pelt, S.; Van Rantwijk, F.; Sheldon, R.
Synthesis of aliphatic (S)-alpha-hydroxycarboxylic amides using a one-pot bienzymatic cascade of immobilised oxynitrilase and nitrile hydratase
Adv. Synth. Catal.
351
397-404
2009
Manihot esculenta
-
Andexer, J.; Guterl, J.K.; Pohl, M.; Eggert, T.
A high-throughput screening assay for hydroxynitrile lyase activity
Chem. Commun. (Camb. )
28
4201-4203
2006
Manihot esculenta (P52705)
Wajant, H.; Pfizenmaier, K.
Identification of potential active-site residues in the hydroxynitrile lyase from Manihot esculenta by site-directed mutagenesis
J. Biol. Chem.
271
25830-25834
1996
Manihot esculenta (P52705), Manihot esculenta
Guterl, J.K.
Andexer, J.N.; Sehl, T.; von Langermann, J.; Frindi-Wosch, I.; Rosenkranz, T.; Fitter, J.; Gruber, K.; Kragl, U.; Eggert, T.; Pohl, M.: Uneven twins: comparison of two enantiocomplementary hydroxynitrile lyases with alpha/beta-hydrolase fold
J. Biotechnol.
141
166-173
2009
Hevea brasiliensis, Manihot esculenta, Manihot esculenta (P52705)
Semba, H.; Ichige, E.; Imanaka, T.; Atomi, H.; Aoyagi, H.
Efficient production of active form recombinant cassava hydroxynitrile lyase using Escherichia coli in low-temperature culture
Methods Mol. Biol.
643
133-144
2010
Manihot esculenta (P52705), Manihot esculenta
Dadashipour, M.; Fukuta, Y.; Asano, Y.
Comparative expression of wild-type and highly soluble mutant His103Leu of hydroxynitrile lyase from Manihot esculenta in prokaryotic and eukaryotic expression systems
Protein Expr. Purif.
77
92-97
2010
Manihot esculenta
Padhi, S.K.; Fujii, R.; Legatt, G.A.; Fossum, S.L.; Berchtold, R.; Kazlauskas, R.J.
Switching from an esterase to a hydroxynitrile lyase mechanism requires only two amino acid substitutions
Chem. Biol.
17
863-871
2010
Hevea brasiliensis, Manihot esculenta
Narayanan, N.N.; Ihemere, U.; Ellery, C.; Sayre, R.T.
Overexpression of hydroxynitrile lyase in cassava roots elevates protein and free amino acids while reducing residual cyanogen levels
PLoS ONE
6
e21996
2011
Manihot esculenta
Asano, Y.; Dadashipour, M.; Yamazaki, M.; Doi, N.; Komeda, H.
Functional expression of a plant hydroxynitrile lyase in Escherichia coli by directed evolution: creation and characterization of highly in vivo soluble mutants
Protein Eng. Des. Sel.
24
607-616
2011
Manihot esculenta (P52705)
Von Langermann, J.; Wapenhensch, S.
Hydroxynitrile lyase-catalyzed synthesis of enantiopure cyanohydrins in Biocatalytic Active Static Emulsions (BASE) with suppression of the non-enzymatic side reaction
Adv. Synth. Catal.
356
2989-2997
2014
Manihot esculenta (P52705)
-
Nakano, S.; Dadashipour, M.; Asano, Y.
Structural and functional analysis of hydroxynitrile lyase from Baliospermum montanum with crystal structure, molecular dynamics and enzyme kinetics
Biochim. Biophys. Acta
1844
2059-2067
2014
Baliospermum montanum (D1MX73), Baliospermum montanum, Manihot esculenta (P52705), Manihot esculenta
Van Rantwijk, F.; Stolz, A.
Enzymatic cascade synthesis of (S)-2-hydroxycarboxylic amides and acids Cascade reactions employing a hydroxynitrile lyase, nitrile-converting enzymes and an amidase
J. Mol. Catal. B
114
25-30
2015
Manihot esculenta (P52705)
-
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