4.2.1.84: nitrile hydratase
This is an abbreviated version!
For detailed information about nitrile hydratase, go to the full flat file.
Word Map on EC 4.2.1.84
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4.2.1.84
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rhodococcus
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amidase
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acrylamide
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rhodochrous
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erythropolis
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synthesis
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feiii
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low-spin
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fe-type
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non-heme
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sulfenic
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benzonitrile
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pseudonocardia
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sulfinate
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propionamide
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cysteine-sulfinic
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neonicotinoid
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ruber
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propionitrile
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cgmcc
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thiacloprid
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carboxamido
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aldoxime
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indole-3-acetonitrile
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chlororaphis
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metallochaperone
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industry
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pharmacology
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degradation
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environmental protection
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analysis
- 4.2.1.84
- rhodococcus
- amidase
- acrylamide
- rhodochrous
- erythropolis
- synthesis
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feiii
-
low-spin
-
fe-type
-
non-heme
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sulfenic
- benzonitrile
- pseudonocardia
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sulfinate
- propionamide
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cysteine-sulfinic
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neonicotinoid
- ruber
- propionitrile
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cgmcc
- thiacloprid
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carboxamido
- aldoxime
- indole-3-acetonitrile
- chlororaphis
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metallochaperone
- industry
- pharmacology
- degradation
- environmental protection
- analysis
Reaction
Synonyms
3-cyanopyridine hydratase, acrylonitrile hydratase, aliphatic nitrile hydratase, ANHase, Co-type NHase, Co-type nitrile hydratase, cobalt-containing nitrile hydratase, CoIII-NHase, CtNHase, Fe-NHase, H-NHase, H-nitrilase, high-molecular mass nitrile hydratase, high-molecular weight nitrile hydratase, hydratase, nitrile, iron-type nitrile hydratase, L-Nhase, L-nitrilase, low-molecular mass nitrile hydratase, low-molecular weight nitrile hydratase, MbNHase, NHase, NHaseK, NI1 NHase, NilCo, NilFe, nitrilase, nitrile hydratase, NthAB, PaNit, ppNHase, ReNHase, TNHase, toyocamycin nitrile hydratase
ECTree
4.2.1.84 nitrile hydrataseAdvanced search results
results (
results (Engineering
Engineering on EC 4.2.1.84 - nitrile hydratase
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
H80A
mutant of alpha-subunit, activity (kcat = 220/s) accounts for about 20% of the wild-type activity (kcat = 1100/s) while the Km value is slightly reduced (187 mM)
H80A/H81A
mutant of alpha-subunit, activity (kcat = 132/s) accounts for 12% of the wild-type activity (kcat = 1100/s) while the Km value is nearly unchanged at 205 mM. Hydrogen-bonding interactions crucial for the catalytic function of the alphaCys104-SOH ligand are disrupted. Disruption of these hydrogen bonding interactions likely alters the nucleophilicity of the sulfenic acid oxygen and the Lewis acidity of the active site Fe(III) ion
H80W/H81W
mutant of alpha-subunit, disruption of these hydrogen bonding interactions likely alters the nucleophilicity of the sulfenic acid oxygen and the Lewis acidity of the active site Fe(III) ion
H81A
mutant of alpha-subunit, activity (kcat = 77/s) accounts for 4% of the wild-type activity (kcat = 1100/s) while the Km value is slightly reduced (179 mM)
R157A
H80A
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mutant of alpha-subunit, activity (kcat = 220/s) accounts for about 20% of the wild-type activity (kcat = 1100/s) while the Km value is slightly reduced (187 mM)
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H80A/H81A
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mutant of alpha-subunit, activity (kcat = 132/s) accounts for 12% of the wild-type activity (kcat = 1100/s) while the Km value is nearly unchanged at 205 mM. Hydrogen-bonding interactions crucial for the catalytic function of the alphaCys104-SOH ligand are disrupted. Disruption of these hydrogen bonding interactions likely alters the nucleophilicity of the sulfenic acid oxygen and the Lewis acidity of the active site Fe(III) ion
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H81A
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mutant of alpha-subunit, activity (kcat = 77/s) accounts for 4% of the wild-type activity (kcat = 1100/s) while the Km value is slightly reduced (179 mM)
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R157A
up
S122A
alpha subunit, site-directed mutagenesis of the recombinant NHase with modified start codon
S122C
alpha subunit, site-directed mutagenesis of the recombinant NHase with modified start codon
S122D
alpha subunit, site-directed mutagenesis of the recombinant NHase with modified start codon
W47E
beta subunit, site-directed mutagenesis of the recombinant NHase with modified start codon
S122A
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alpha subunit, site-directed mutagenesis of the recombinant NHase with modified start codon
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S122C
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alpha subunit, site-directed mutagenesis of the recombinant NHase with modified start codon
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S122D
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alpha subunit, site-directed mutagenesis of the recombinant NHase with modified start codon
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W47E
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beta subunit, site-directed mutagenesis of the recombinant NHase with modified start codon
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M150C
beta-subunit mutant enzyme, 32% increase in half-life at 50°C, the kcat/Km value is 1.1fold higher than the kcat/Km value of the wild-type enzyme
S189E
beta-subunit mutant enzyme, 107% increase in half-life at 50°C, the kcat/Km value is 2.2fold higher than the kcat/Km value of the wild-type enzyme
T173Y
beta-subunit mutant enzyme, 7% increase in half-life at 50°C, the kcat/Km value is 1.5fold higher than the kcat/Km value of the wild-type enzyme
M150C
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beta-subunit mutant enzyme, 32% increase in half-life at 50°C, the kcat/Km value is 1.1fold higher than the kcat/Km value of the wild-type enzyme
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S189E
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beta-subunit mutant enzyme, 107% increase in half-life at 50°C, the kcat/Km value is 2.2fold higher than the kcat/Km value of the wild-type enzyme
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T173Y
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beta-subunit mutant enzyme, 7% increase in half-life at 50°C, the kcat/Km value is 1.5fold higher than the kcat/Km value of the wild-type enzyme
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alphaT109S
site-directed mutagenesis, the mutant shows similar characteristics compared to the wild-type enzyme
alphaY114T
site-directed mutagenesis, the mutant shows a very low cobalt content and catalytic activity compared to the wild-type enzyme, and oxidative modifications of aCys111 and aCys113 residues are not observed
betaY68F
site-directed mutagenesis, the mutant shows an elevated Km value and a significantly decreased kcat value compared to the wild-type enzyme
T109S
similar characteristics to the wild-type enzyme
Y114T
very low cobalt content and catalytic activity compared to the wild-type enzyme
alphaT109S
-
site-directed mutagenesis, the mutant shows similar characteristics compared to the wild-type enzyme
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alphaY114T
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site-directed mutagenesis, the mutant shows a very low cobalt content and catalytic activity compared to the wild-type enzyme, and oxidative modifications of aCys111 and aCys113 residues are not observed
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betaY68F
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site-directed mutagenesis, the mutant shows an elevated Km value and a significantly decreased kcat value compared to the wild-type enzyme
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Y114T
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very low cobalt content and catalytic activity compared to the wild-type enzyme
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S113A
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the mutation partially affects catalytic activity and does not change the pH profiles of the kinetic parameters, the electronic state of the Fe center is altered
S113A
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the mutation partially affects catalytic activity and does not change the pH profiles of the kinetic parameters, the electronic state of the Fe center is altered
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A51L
wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 74.3%
alphaV5L
-
site-directed mutagenesis, exchange in the H-NHase does not influence the catalytic activity or the Co2+ content
F37H
wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 96.8%
F37H/F51L
wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 94.6%
F37H/L48A
wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 95.1%
F37Y
wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 88.7%
F51Q
wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 60.2%
L48Q
wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 59.7%
W72Y
with adiponitrile as substrate the wild-type enzyme forms only adipamide after a 4 h reaction. Y68T and W72Y mutations cause a significant shift in product formation and form primarily 5-cyanovaleramide. The use of the wild-type enzyme leads to malonamide formation with 97.3% malononitrile conversion. Variants Y68T and W72Y show a drastic change in regiospecificity by producing mainly the omega-cyanocarboxamide, cyanoacetamide, at a relatively low malononitrile conversion. The wild-type enzyme prefers terephthalonitrile to catalyze mainly into terephthalamide (84.3%) with a conversion up to 99.2 %. Variants Y68T and W72Y show a change in regiospecificity by producing mainly the 4-cyanobenzamide. The wild-type enzyme forms 100% phthalamide from phthalodinitrile. The mutant enzymes Y68T and W72Y result in a higher 2-cyanobenzamide formation than their parent enzyme
Y68T
with adiponitrile as substrate the wild-type enzyme forms only adipamide after a 4 h reaction. Y68T and W72Y mutations cause a significant shift in product formation and form primarily 5-cyanovaleramide. The use of the wild-type enzyme leads to malonamide formation with 97.3% malononitrile conversion. Variants Y68T and W72Y show a drastic change in regiospecificity by producing mainly the omega-cyanocarboxamide, cyanoacetamide, at a relatively low malononitrile conversion. The wild-type enzyme prefers terephthalonitrile to catalyze mainly into terephthalamide (84.3%) with a conversion up to 99.2%. Variants Y68T and W72Y show a change in regiospecificity by producing mainly the 4-cyanobenzamide. The wild-type enzyme forms 100% phthalamide from phthalodinitrile. The mutant enzymes Y68T and W72Y result in a higher 2-cyanobenzamide formation than their parent enzyme
Y68T/W72Y
the mutant enzyme displays a totally different regioselectivity towards dinitriles than its parent enzyme. The use of the wild-type enzyme leads to malonamide formation with 97.3% malononitrile conversion. Mutant enzyme Y68T/W72Y produces 97.1% omega-cyanocarboxamide. The wild-type enzyme forms only adipamide after a 4 h reaction. Mutant enzyme Y68T/W72Y produces 100% 5-cyanovaleramide. The wild-type enzyme prefers terephthalonitrile to catalyze mainly into terephthalamide (84.3%) with a conversion up to 99.2 %. Mutant enzyme Y68T/W72Y produces 98.2% 4-cyanobenzamide from terephthalonitrile. The wild-type enzyme forms 100% phthalamide from phthalodinitrile. Mutant enzyme Y68T/W72Y produces 100% 2-cyanobenzamide from phthalodinitrile
A51L
-
wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 74.3%
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alphaV5L
-
site-directed mutagenesis, exchange in the H-NHase does not influence the catalytic activity or the Co2+ content
-
F37H
-
wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 96.8%
-
F37Y
-
wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 88.7%
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F51Q
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wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 60.2%
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L48Q
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wild-type enzyme catalyzes the conversion of (S)-mandelamide with an enantiomeric excess of 52.6%, the beta-subunit mutant enzyme catalyzes the reaction with an enantiomeric excess of 59.7%
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W72Y
-
with adiponitrile as substrate the wild-type enzyme forms only adipamide after a 4 h reaction. Y68T and W72Y mutations cause a significant shift in product formation and form primarily 5-cyanovaleramide. The use of the wild-type enzyme leads to malonamide formation with 97.3% malononitrile conversion. Variants Y68T and W72Y show a drastic change in regiospecificity by producing mainly the omega-cyanocarboxamide, cyanoacetamide, at a relatively low malononitrile conversion. The wild-type enzyme prefers terephthalonitrile to catalyze mainly into terephthalamide (84.3%) with a conversion up to 99.2 %. Variants Y68T and W72Y show a change in regiospecificity by producing mainly the 4-cyanobenzamide. The wild-type enzyme forms 100% phthalamide from phthalodinitrile. The mutant enzymes Y68T and W72Y result in a higher 2-cyanobenzamide formation than their parent enzyme
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Y68T
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with adiponitrile as substrate the wild-type enzyme forms only adipamide after a 4 h reaction. Y68T and W72Y mutations cause a significant shift in product formation and form primarily 5-cyanovaleramide. The use of the wild-type enzyme leads to malonamide formation with 97.3% malononitrile conversion. Variants Y68T and W72Y show a drastic change in regiospecificity by producing mainly the omega-cyanocarboxamide, cyanoacetamide, at a relatively low malononitrile conversion. The wild-type enzyme prefers terephthalonitrile to catalyze mainly into terephthalamide (84.3%) with a conversion up to 99.2%. Variants Y68T and W72Y show a change in regiospecificity by producing mainly the 4-cyanobenzamide. The wild-type enzyme forms 100% phthalamide from phthalodinitrile. The mutant enzymes Y68T and W72Y result in a higher 2-cyanobenzamide formation than their parent enzyme
-
Y68T/W72Y
-
the mutant enzyme displays a totally different regioselectivity towards dinitriles than its parent enzyme. The use of the wild-type enzyme leads to malonamide formation with 97.3% malononitrile conversion. Mutant enzyme Y68T/W72Y produces 97.1% omega-cyanocarboxamide. The wild-type enzyme forms only adipamide after a 4 h reaction. Mutant enzyme Y68T/W72Y produces 100% 5-cyanovaleramide. The wild-type enzyme prefers terephthalonitrile to catalyze mainly into terephthalamide (84.3%) with a conversion up to 99.2 %. Mutant enzyme Y68T/W72Y produces 98.2% 4-cyanobenzamide from terephthalonitrile. The wild-type enzyme forms 100% phthalamide from phthalodinitrile. Mutant enzyme Y68T/W72Y produces 100% 2-cyanobenzamide from phthalodinitrile
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betaW47E
additional information
R157A
mutant of alpha-subunit, activity (kcat = 10/s) accounts for less than 1% of the wild-type activity (kcat = 1100/s) while the Km value is nearly unchanged at 205 mM
R157A
mutant of alpha-subunit, disruption of these hydrogen bonding interactions likely alters the nucleophilicity of the sulfenic acid oxygen and the Lewis acidity of the active site Fe(III) ion
R157A
-
mutant of alpha-subunit, activity (kcat = 10/s) accounts for less than 1% of the wild-type activity (kcat = 1100/s) while the Km value is nearly unchanged at 205 mM
-
R157A
-
mutant of alpha-subunit, disruption of these hydrogen bonding interactions likely alters the nucleophilicity of the sulfenic acid oxygen and the Lewis acidity of the active site Fe(III) ion
-
up
the putative activator (17K) gene next to the beta-subunit gene is proven to be important for the functional expression of recombinant NHaseK in Escherichia coli
up
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the putative activator (17K) gene next to the beta-subunit gene is proven to be important for the functional expression of recombinant NHaseK in Escherichia coli
-
additional information
-
improvement of thermal stability of the industrialized mesophilic NHase by introducing stable salt-bridge interactions into its thermal sensitive regions
additional information
transfer of stabilized salt-bridge interactions from three deformation-prone thermal-sensitive regions (A1, A2 and A3 in beta-subunit) of the thermophilic NHase 1V29 from Bacillus SC-105-1 and 1UGQ from Pseudonocardia thermophila JCM3095 into industrialized mesophilic NHase-TH from Rhodococcus ruber TH
additional information
transfer of stabilized salt-bridge interactions from three deformation-prone thermal-sensitive regions (A1, A2 and A3 in beta-subunit) of the thermophilic NHase 1V29 from Bacillus SC-105-1 and 1UGQ from Pseudonocardia thermophila JCM3095 into industrialized mesophilic NHase-TH from Rhodococcus ruber TH
additional information
Bacillus sp. (in: Bacteria) SC-105-1
-
improvement of thermal stability of the industrialized mesophilic NHase by introducing stable salt-bridge interactions into its thermal sensitive regions
-
additional information
Bacillus sp. (in: Bacteria) SC-105-1
-
transfer of stabilized salt-bridge interactions from three deformation-prone thermal-sensitive regions (A1, A2 and A3 in beta-subunit) of the thermophilic NHase 1V29 from Bacillus SC-105-1 and 1UGQ from Pseudonocardia thermophila JCM3095 into industrialized mesophilic NHase-TH from Rhodococcus ruber TH
-
additional information
the chimeric NHase (SBpNHase) from the thermal sensitive nitrile hydratasese from Bordetella petrii and the relatively thermal-stable nitrile hydratases from Pseudonocardia thermophila is constructed by swapping the corresponding C-domains
additional information
-
the chimeric NHase (SBpNHase) from the thermal sensitive nitrile hydratasese from Bordetella petrii and the relatively thermal-stable nitrile hydratases from Pseudonocardia thermophila is constructed by swapping the corresponding C-domains
additional information
Bordetella petrii DSM 128043
-
the chimeric NHase (SBpNHase) from the thermal sensitive nitrile hydratasese from Bordetella petrii and the relatively thermal-stable nitrile hydratases from Pseudonocardia thermophila is constructed by swapping the corresponding C-domains
-
additional information
-
Fe of an Fe-dependent recombinant nitrile hydratase (wild-type, NilFe) is replaced by Co generating an Co-substituted enzyme (NilCo)
additional information
homologous protein fragment swapping method is used for the improvement of the stability of NHase from Pseudomonas putida NRRL-18668, site targeted amino recombination software and molecular dynamics are used for determination of the crossover sites for fragment recombination, overview. One thermophilic NHase fragment M1 to G98 from Comamonas testosteroni 5-MGAM-4D and two fragments K165-V196 and K165-D209 from Pseudonocardia thermophila JCM3095 are selected to swap the corresponding fragments of Pseudomonas putida NHase. The chimeric NHases show 1.4 to 3.5fold enhancement in thermostability, some show reduced activity and product inhibition compared to wild-type Pseudomonas putida NHase. But mutants 3AB and 3ABC show increased activity due to altered secondary structure compared to the Pseudomonas putida wild-type
additional information
homologous protein fragment swapping method is used for the improvement of the stability of NHase from Pseudomonas putida NRRL-18668, site targeted amino recombination software and molecular dynamics are used for determination of the crossover sites for fragment recombination, overview. One thermophilic NHase fragment M1 to G98 from Comamonas testosteroni 5-MGAM-4D and two fragments K165-V196 and K165-D209 from Pseudonocardia thermophila JCM3095 are selected to swap the corresponding fragments of Pseudomonas putida NHase. The chimeric NHases show 1.4 to 3.5fold enhancement in thermostability, some show reduced activity and product inhibition compared to wild-type Pseudomonas putida NHase. But mutants 3AB and 3ABC show increased activity due to altered secondary structure compared to the Pseudomonas putida wild-type
additional information
-
homologous protein fragment swapping method is used for the improvement of the stability of NHase from Pseudomonas putida NRRL-18668, site targeted amino recombination software and molecular dynamics are used for determination of the crossover sites for fragment recombination, overview. One thermophilic NHase fragment M1 to G98 from Comamonas testosteroni 5-MGAM-4D and two fragments K165-V196 and K165-D209 from Pseudonocardia thermophila JCM3095 are selected to swap the corresponding fragments of Pseudomonas putida NHase. The chimeric NHases show 1.4 to 3.5fold enhancement in thermostability, some show reduced activity and product inhibition compared to wild-type Pseudomonas putida NHase. But mutants 3AB and 3ABC show increased activity due to altered secondary structure compared to the Pseudomonas putida wild-type
-
additional information
-
homologous protein fragment swapping method is used for the improvement of the stability of NHase from Pseudomonas putida NRRL-18668, site targeted amino recombination software and molecular dynamics are used for determination of the crossover sites for fragment recombination, overview. One thermophilic NHase fragment M1 to G98 from Comamonas testosteroni 5-MGAM-4D and two fragments K165-V196 and K165-D209 from Pseudonocardia thermophila JCM3095 are selected to swap the corresponding fragments of Pseudomonas putida NHase. The chimeric NHases show 1.4 to 3.5fold enhancement in thermostability, some show reduced activity and product inhibition compared to wild-type Pseudomonas putida NHase. But mutants 3AB and 3ABC show increased activity due to altered secondary structure compared to the wild-type
additional information
-
the thermostability and product tolerance of nitrile hydratase are enhanced by fusing with two of the self-assembling amphipathic peptides, EAK16 (AEAEAKAKAEAEAKAK) at the N- and C-terminus and ELK16 (LELELKLKLELELKLK) at the N-terminus. When self-assembling amphipathic peptide ELK16 is fused to the N-terminus of the enzymes beta-subunit, the resultant enzyme (SAP-NHase-2) becomes an active inclusion body, while EAK16 does not affect the enzyme solubility when fused to the enzyme's C-terminus (SAP-NHase-10) or N-termninus (AP-NHase-1) of the beta-subunit
additional information
-
homologous protein fragment swapping method is used for the improvement of the stability of NHase from Pseudomonas putida NRRL-18668, site targeted amino recombination software and molecular dynamics are used for determination of the crossover sites for fragment recombination, overview. One thermophilic NHase fragment M1 to G98 from Comamonas testosteroni 5-MGAM-4D and two fragments K165-V196 and K165-D209 from Pseudonocardia thermophila JCM3095 are selected to swap the corresponding fragments of Pseudomonas putida NHase. The chimeric NHases show 1.4 to 3.5fold enhancement in thermostability, some show reduced activity and product inhibition compared to wild-type Pseudomonas putida NHase. But mutants 3AB and 3ABC show increased activity due to altered secondary structure compared to the wild-type
-
additional information
-
the thermostability and product tolerance of nitrile hydratase are enhanced by fusing with two of the self-assembling amphipathic peptides, EAK16 (AEAEAKAKAEAEAKAK) at the N- and C-terminus and ELK16 (LELELKLKLELELKLK) at the N-terminus. When self-assembling amphipathic peptide ELK16 is fused to the N-terminus of the enzymes beta-subunit, the resultant enzyme (SAP-NHase-2) becomes an active inclusion body, while EAK16 does not affect the enzyme solubility when fused to the enzyme's C-terminus (SAP-NHase-10) or N-termninus (AP-NHase-1) of the beta-subunit
-
additional information
improvement of thermal stability of the industrialized mesophilic NHase by introducing stable salt-bridge interactions into its thermal sensitive regions
additional information
homologous protein fragment swapping method is used for the improvement of the stability of NHase from Pseudomonas putida NRRL-18668, site targeted amino recombination software and molecular dynamics are used for determination of the crossover sites for fragment recombination, overview. One thermophilic NHase fragment M1 to G98 from Comamonas testosteroni 5-MGAM-4D and two fragments K165-V196 and K165-D209 from Pseudonocardia thermophila JCM3095 are selected to swap the corresponding fragments of Pseudomonas putida NHase. The chimeric NHases show 1.4 to 3.5fold enhancement in thermostability, some show reduced activity and product inhibition compared to wild-type Pseudomonas putida NHase. But mutants 3AB and 3ABC show increased activity due to altered secondary structure compared to the Pseudomonas putida wild-type
additional information
homologous protein fragment swapping method is used for the improvement of the stability of NHase from Pseudomonas putida NRRL-18668, site targeted amino recombination software and molecular dynamics are used for determination of the crossover sites for fragment recombination, overview. One thermophilic NHase fragment M1 to G98 from Comamonas testosteroni 5-MGAM-4D and two fragments K165-V196 and K165-D209 from Pseudonocardia thermophila JCM3095 are selected to swap the corresponding fragments of Pseudomonas putida NHase. The chimeric NHases show 1.4 to 3.5fold enhancement in thermostability, some show reduced activity and product inhibition compared to wild-type Pseudomonas putida NHase. But mutants 3AB and 3ABC show increased activity due to altered secondary structure compared to the Pseudomonas putida wild-type
additional information
-
transfer of stabilized salt-bridge interactions from three deformation-prone thermal-sensitive regions (A1, A2 and A3 in beta-subunit) of the thermophilic NHase 1V29 from Bacillus SC-105-1 and 1UGQ from Pseudonocardia thermophila JCM3095 into industrialized mesophilic NHase-TH from Rhodococcus ruber TH
additional information
the chimeric NHase (SBpNHase) from the thermal sensitive nitrile hydratasese from Bordetella petrii and the relatively thermal-stable nitrile hydratases from Pseudonocardia thermophila is constructed by swapping the corresponding C-domains
additional information
-
transfer of stabilized salt-bridge interactions from three deformation-prone thermal-sensitive regions (A1, A2 and A3 in beta-subunit) of the thermophilic NHase 1V29 from Bacillus SC-105-1 and 1UGQ from Pseudonocardia thermophila JCM3095 into industrialized mesophilic NHase-TH from Rhodococcus ruber TH
-
additional information
-
homologous protein fragment swapping method is used for the improvement of the stability of NHase from Pseudomonas putida NRRL-18668, site targeted amino recombination software and molecular dynamics are used for determination of the crossover sites for fragment recombination, overview. One thermophilic NHase fragment M1 to G98 from Comamonas testosteroni 5-MGAM-4D and two fragments K165-V196 and K165-D209 from Pseudonocardia thermophila JCM3095 are selected to swap the corresponding fragments of Pseudomonas putida NHase. The chimeric NHases show 1.4 to 3.5fold enhancement in thermostability, some show reduced activity and product inhibition compared to wild-type Pseudomonas putida NHase. But mutants 3AB and 3ABC show increased activity due to altered secondary structure compared to the Pseudomonas putida wild-type
-
additional information
-
improvement of thermal stability of the industrialized mesophilic NHase by introducing stable salt-bridge interactions into its thermal sensitive regions
-
additional information
-
construction of an amidase-negative, amiE-, recombinant strain TH3 with 25% increased nitrile hydratase activity and 60% reduced amidase activity compared to the wild-type strain TH. Usage of TH3 free cells as biocatalysts at 18°C for acrylamide production, increased by 23% and 87% reduced acrylic acid by-product formation conmpared to the wild-type
additional information
-
transfer of stabilized salt-bridge interactions from three deformation-prone thermal-sensitive regions (A1, A2 and A3 in beta-subunit) of the thermophilic NHase 1V29 from Bacillus SC-105-1 and 1UGQ from Pseudonocardia thermophila JCM3095 into industrialized mesophilic NHase-TH from Rhodococcus ruber TH. Three types of salt bridge - active center-adjacent (in A1), internal neighboring-residue-bridged (in A2) and C-terminal-residue-bridged (A3) - are constructed in NHase-TH. A C-terminal salt-bridge strategy is powerful for enzyme stability intensification through triggering global changes of the salt bridge networks, molecular dynamic simulation, overview
additional information
-
construction of an amidase-negative, amiE-, recombinant strain TH3 with 25% increased nitrile hydratase activity and 60% reduced amidase activity compared to the wild-type strain TH. Usage of TH3 free cells as biocatalysts at 18°C for acrylamide production, increased by 23% and 87% reduced acrylic acid by-product formation conmpared to the wild-type
-
additional information
-
transfer of stabilized salt-bridge interactions from three deformation-prone thermal-sensitive regions (A1, A2 and A3 in beta-subunit) of the thermophilic NHase 1V29 from Bacillus SC-105-1 and 1UGQ from Pseudonocardia thermophila JCM3095 into industrialized mesophilic NHase-TH from Rhodococcus ruber TH. Three types of salt bridge - active center-adjacent (in A1), internal neighboring-residue-bridged (in A2) and C-terminal-residue-bridged (A3) - are constructed in NHase-TH. A C-terminal salt-bridge strategy is powerful for enzyme stability intensification through triggering global changes of the salt bridge networks, molecular dynamic simulation, overview
-
