3.8.1.5: haloalkane dehalogenase
This is an abbreviated version!
For detailed information about haloalkane dehalogenase, go to the full flat file.
Word Map on EC 3.8.1.5
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3.8.1.5
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xanthobacter
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autotrophicus
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dehalogenation
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1,2-dichloroethane
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halide
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carbon-halogen
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1,2-dibromoethane
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sphingomonas
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paucimobilis
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1,2,3-trichloropropane
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synthesis
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hexachlorocyclohexane
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rhodochrous
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sphingobium
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environmental protection
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alkyl-enzyme
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haloacid
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1-chlorobutane
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2-chloroethanol
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ncimb
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chloroalkane
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dehydrochlorinase
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epichlorohydrine
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gamma-hexachlorocyclohexane
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halotag
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biotechnology
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alpha/beta-hydrolase
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haloalcohols
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agriculture
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halide-binding
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degradation
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industry
- 3.8.1.5
- xanthobacter
- autotrophicus
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dehalogenation
- 1,2-dichloroethane
- halide
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carbon-halogen
- 1,2-dibromoethane
- sphingomonas
- paucimobilis
- 1,2,3-trichloropropane
- synthesis
- hexachlorocyclohexane
- rhodochrous
- sphingobium
- environmental protection
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alkyl-enzyme
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haloacid
- 1-chlorobutane
- 2-chloroethanol
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ncimb
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chloroalkane
- dehydrochlorinase
- epichlorohydrine
- gamma-hexachlorocyclohexane
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halotag
- biotechnology
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alpha/beta-hydrolase
- haloalcohols
- agriculture
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halide-binding
- degradation
- industry
Reaction
Synonyms
1,3,4,6,-tetrachloro-1,4-cyclohexadiene halidohydrolase, 1-chlorohexane halidohydrolase, 1-haloalkane dehalogenase, DadB, DatA, DbeA, DbjA, DccA, DhaA, DhaA31, DhaB, DhaC, DhAf, DhlA, DhmA, DmaA, dmbA, DmbB, DmbC, dmlA, DmmA, DmrA, DmrB, DmsA, DmtA, DmxA, DpcA, DppA, DrbA, DsaA, DspA, EC 3.8.1.1, eHLD-B, eHLD-C, haloalkane dehalogenase, haloalkane dehalogenase LinB, HanR, HLD, HLD-I, LinB, LinBMI, LinBUT, metallo-haloalkane dehalogenase, protein XP_504164, Rv2579, Ylehd
ECTree
Advanced search results
Engineering
Engineering on EC 3.8.1.5 - haloalkane dehalogenase
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Y109W
D103A
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no activity with substrates 1,3-dibromopropane or 1-chlorobutane
E127A
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no activity with substrates 1,3-dibromopropane or 1-chlorobutane, purification of enzyme not possible because of very small amount of soluble enzyme protein
H280A
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no activity with substrates 1,3-dibromopropane or 1-chlorobutane
D103A
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no activity with substrates 1,3-dibromopropane or 1-chlorobutane
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E127A
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no activity with substrates 1,3-dibromopropane or 1-chlorobutane, purification of enzyme not possible because of very small amount of soluble enzyme protein
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H280A
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no activity with substrates 1,3-dibromopropane or 1-chlorobutane
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D123A
D250A
H279A
W124L
W164L
C176Y/Y273F
W118F
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mutant with increased KM and decreased kcat for primary, secondary and cyclic alkylhalide substrates
F168W/A172L/Y176G
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site-directed mutagenesis, the mutant shows increased enantioselectivity with substrate TCP compared to the wild-type enzyme, 1,2,3-trichloropropane is docked in the active site in a configuration that leads to (R)-2,3-dichloropropan-1-ol formation
I135F/C176Y/V245F/L246I/Y273F
C176Y
A247H
mutant, constructed for quantitative evaluation of the differences between LinB of Sphingobium japonicum and LinB of Sphingobium sp.
I134A
mutant, constructed for quantitative evaluation of the differences between LinB of Sphingobium japonicum and LinB of Sphingobium sp.
I134V/A247H
mutant, constructed for quantitative evaluation of the differences between LinB of Sphingobium japonicum and LinB of Sphingobium sp.
L177W
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naturally occuring mutation, the single mutation in a tunnel to the active site changes the mechanism and kinetics of product release in LinB. Interactions of the bromide ion with the tryptophan increase free energy barrier for its passage, causing the reaction mechanism change
A247H
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mutant, constructed for quantitative evaluation of the differences between LinB of Sphingobium japonicum and LinB of Sphingobium sp.
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I134A
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mutant, constructed for quantitative evaluation of the differences between LinB of Sphingobium japonicum and LinB of Sphingobium sp.
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I134V/A247H
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mutant, constructed for quantitative evaluation of the differences between LinB of Sphingobium japonicum and LinB of Sphingobium sp.
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L177W
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naturally occuring mutation, the single mutation in a tunnel to the active site changes the mechanism and kinetics of product release in LinB. Interactions of the bromide ion with the tryptophan increase free energy barrier for its passage, causing the reaction mechanism change
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H247A
V134I
V134I/H247A
mutant, constructed for quantitative evaluation of the differences between LinB of Sphingobium japonicum and LinB of Sphingobium sp.
H247A
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mutant, constructed for quantitative evaluation of the differences between LinB of Sphingobium japonicum and LinB of Sphingobium sp.
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V134I
V134I/H247A
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mutant, constructed for quantitative evaluation of the differences between LinB of Sphingobium japonicum and LinB of Sphingobium sp.
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F151L
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site-directed mutagenesis, altered catalytic properties compared to the wild-type enzyme
F151W
F151Y
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site-directed mutagenesis, altered catalytic properties compared to the wild-type enzyme
F169L
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site-directed mutagenesis, altered catalytic properties compared to the wild-type enzyme
L177A
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site-directed mutagenesis, slightly reduced activity compared to the wild-type enzyme
L177C
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site-directed mutagenesis, altered substrate specificity, especially high activity with 1-iodobutane, bromocyclohexane and 1-bromobutane
L177D
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site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme
L177E
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site-directed mutagenesis, mutant cannot be expressed in Escherichia coli due to incorrect protein folding
L177F
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site-directed mutagenesis, slightly reduced activity compared to the wild-type enzyme
L177H
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site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme
L177I
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site-directed mutagenesis, inactive mutant, incorrect protein folding
L177M
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site-directed mutagenesis, slightly reduced activity compared to the wild-type enzyme
L177N
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site-directed mutagenesis, mutant cannot be expressed in Escherichia coli due to incorrect protein folding
L177P
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site-directed mutagenesis, inactive mutant, incorrect protein folding
L177Q
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site-directed mutagenesis, altered substrate specificity, especially high activity with 1-iodobutane and 1-bromobutane
L177R
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site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme
L177S
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site-directed mutagenesis, altered substrate specificity, especially high activity with 1-iodobutane and 1-bromobutane
L177T
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site-directed mutagenesis, altered substrate specificity, especially high activity with 1-iodobutane and 1-bromobutane
L177V
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site-directed mutagenesis, altered substrate specificity, especially high activity with 1-bromobutane
L177W
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site-directed mutagenesis, altered substrate specificity, specifically highly active with 1-chlorobutane
L177Y
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site-directed mutagenesis, altered substrate specificity, especially low activity with 1,2-dibromoethane
N38D
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site-directed mutagenesis, altered catalytic properties compared to the wild-type enzyme
N38E
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site-directed mutagenesis, altered catalytic properties compared to the wild-type enzyme
N38F
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site-directed mutagenesis, altered catalytic properties compared to the wild-type enzyme
N38Q
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site-directed mutagenesis, altered catalytic properties compared to the wild-type enzyme
W109L
F151W
L177A
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site-directed mutagenesis, slightly reduced activity compared to the wild-type enzyme
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L177E
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site-directed mutagenesis, mutant cannot be expressed in Escherichia coli due to incorrect protein folding
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L177I
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site-directed mutagenesis, inactive mutant, incorrect protein folding
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L177N
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site-directed mutagenesis, mutant cannot be expressed in Escherichia coli due to incorrect protein folding
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L177P
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site-directed mutagenesis, inactive mutant, incorrect protein folding
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N38D
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site-directed mutagenesis, altered catalytic properties compared to the wild-type enzyme
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N38E
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site-directed mutagenesis, altered catalytic properties compared to the wild-type enzyme
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N38F
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site-directed mutagenesis, altered catalytic properties compared to the wild-type enzyme
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W109L
F172W
F294A
mutant with modified kinetic mechanism for halide binding
H289Q
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GJ10, mutant with 660fold reduced activity with substrate 1,2-dibromoethane
T197A
mutant with modified kinetic mechanism for halide binding
W175Y
F172W
H289Q
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GJ10, mutant with 660fold reduced activity with substrate 1,2-dibromoethane
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W175Y
additional information
site-directed mutagenesis, crystal structure determination
Y109W
site-directed mutagenesis, structure and catalytic properties by spectroscopy and pre-steady-state kinetic experiments, quantum mechanical and molecular dynamics calculations. The mutation affects the substrate specificity of the enzyme and reduced its KM for selected halogenated substrates by a factor of 2-4, without impacting the rate-determining hydrolytic step
Y109W
Agrobacterium tumefaciens C58 / ATCC 33970
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site-directed mutagenesis, crystal structure determination
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Y109W
Agrobacterium tumefaciens C58 / ATCC 33970
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site-directed mutagenesis, structure and catalytic properties by spectroscopy and pre-steady-state kinetic experiments, quantum mechanical and molecular dynamics calculations. The mutation affects the substrate specificity of the enzyme and reduced its KM for selected halogenated substrates by a factor of 2-4, without impacting the rate-determining hydrolytic step
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D123A
mutant to verify catalytic relevant residues, no statistically significant activity is detected
D250A
mutant to verify catalytic relevant residues, no statistically significant activity is detected
H279A
mutant to verify catalytic relevant residues, no statistically significant activity is detected
W124L
mutant to verify catalytic relevant residues, no statistically significant activity is detected
W164L
mutant to verify catalytic relevant residues, no statistically significant activity is detected
site-directed mutagenesis, the mutant enzyme is 3.5fold higher activity toward TCP than DhaA wild-type
C176Y/Y273F
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site-directed mutagenesis, the mutant enzyme is 3.5fold higher activity toward TCP than DhaA wild-type
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structural modeling of the mutant compared to the wild-type DhaA31 using the wild-type crystal structure
I135F/C176Y/V245F/L246I/Y273F
four out of five mutations are located in the tunnels and introduce bulkier residues, which narrowed the p1 and p2 tunnels leading to its active site. The mutant enzyme has higher activity toward 1,2,3-trichloropropane than the wild-type, with enhancements of 32fold on the catalytic constant kcat and 26fold on the catalytic efficiency (kcat/Km). The carbon-halogen bond is the rate-limiting step in the wild-type. This step is enhanced in DhaA31 due to a significantly higher number of reactive configurations of the substrate and a decrease of the energy barrier to the SN2 reaction. C176Y and V245F are identified as the key mutations responsible for most of those improvements. The release of the alcohol product is the rate-limiting step in the mutant enzyme primarily due to the C176Y mutation. Mutational dissection of the mutant and kinetic analysis of the intermediate mutants confirm the theoretical observations
threefold increase in catalytic efficiency for substrate 1,2,3-trichloropropane. Mutation modifies the protein access and export routes
C176Y
mutant with a threefold higher catalytic efficiency for 1,2,3-trichloropropane dehalogenation
H247A
mutant, constructed for quantitative evaluation of the differences between LinB of Sphingobium japonicum and LinB of Sphingobium sp.
V134I
mutant, constructed for quantitative evaluation of the differences between LinB of Sphingobium japonicum and LinB of Sphingobium sp.
V134I
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mutant, constructed for quantitative evaluation of the differences between LinB of Sphingobium japonicum and LinB of Sphingobium sp.
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F151W
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site-directed mutagenesis, altered catalytic properties compared to the wild-type enzyme
W109L
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site-directed mutagenesis, altered catalytic properties compared to the wild-type enzyme
F151W
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site-directed mutagenesis, altered catalytic properties compared to the wild-type enzyme
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W109L
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site-directed mutagenesis, altered catalytic properties compared to the wild-type enzyme
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mutant with 10fold higher KM for 1-chlorohexane and 2fold higher KM for 1,2-dibromoethane, increased bromide release but velocity of hydrolysis of alkyl-enzyme bond remains unchanged
F172W
steady state kinetics of the mutant compared to the wild-type enzyme
F172W
the conformational change of the mutant enzyme is faster than that of the wild-type enzyme. The open conformation is more common than the closed one. The closed conformation also displays a larger distance between the two helices than what is observed in the wild-type simulations
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mutant with 10fold higher KM for 1-chlorohexane and 2fold higher KM for 1,2-dibromoethane, increased bromide release but velocity of hydrolysis of alkyl-enzyme bond remains unchanged
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F172W
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the conformational change of the mutant enzyme is faster than that of the wild-type enzyme. The open conformation is more common than the closed one. The closed conformation also displays a larger distance between the two helices than what is observed in the wild-type simulations
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F172W
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steady state kinetics of the mutant compared to the wild-type enzyme
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construction of deletion mutant DbeADELTACl, mutation in the second halide-binding site
additional information
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construction of deletion mutant DbeADELTACl, mutation in the second halide-binding site
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additional information
construction of deletion mutants DbjADELTA and DbjADELTA H139A
additional information
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construction of deletion mutants DbjADELTA and DbjADELTA H139A
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additional information
construction of engineered enzyme variants: the more thermostable Dhla5/Dhla8, the more active DhaAM2(C176Y/Y273F), the DhaA31 mutant that is 32fold higher activity toward TCP than DhaA wild-type, te mutant DhaA12 mimicking DbjA active (insertion of ERB), and the DhaA31 mutant that has higher enantioselectivity toward TCP than the wild-type
additional information
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construction of engineered enzyme variants: the more thermostable Dhla5/Dhla8, the more active DhaAM2(C176Y/Y273F), the DhaA31 mutant that is 32fold higher activity toward TCP than DhaA wild-type, te mutant DhaA12 mimicking DbjA active (insertion of ERB), and the DhaA31 mutant that has higher enantioselectivity toward TCP than the wild-type
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additional information
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library screening for mutants with altered enantioselectivity with substrate 1,2,3-trichloropropane compared to DhaA31 wild-type, overview
additional information
development and evaluation of a high-throughput system to select active haloalkane dehalogenase variants from a large mutant library, enrichment of the active wild-type enzyme in contrast to the inactive variants is about 340fold. Three saturation libraries, with a size of more than 106 cells, based on inactive variants of the haloalkane dehalogenases DhaA are successfully screened to retrieve active enzymes
additional information
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construction of LinBUT variants, 4H7D/4H7E/4H7F/4H7H/4H7I/4H7J/4H7K are intermediates between LinBUT and LinBMI
additional information
P51698
construction of LinBUT variants, 4H7D/4H7E/4H7F/4H7H/4H7I/4H7J/4H7K are intermediates between LinBUT and LinBMI
additional information
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construction of LinBUT variants, 4H7D/4H7E/4H7F/4H7H/4H7I/4H7J/4H7K are intermediates between LinBUT and LinBMI
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additional information
construction of LinB variants, that mimick the DmtA active site, shows higher activity toward beta- and delta-HCH than LinBMI. Mutant 4H7D/4H7E/4H7F/4H7H/4H7I/4H7J/4H7K are intermediates between LinBUT and LinBMI
additional information
mutations are constructed corresponding to the residues in LinBUT from Sphingobium japonicum strain UT26
additional information
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construction of LinB variants, that mimick the DmtA active site, shows higher activity toward beta- and delta-HCH than LinBMI. Mutant 4H7D/4H7E/4H7F/4H7H/4H7I/4H7J/4H7K are intermediates between LinBUT and LinBMI
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additional information
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mutations are constructed corresponding to the residues in LinBUT from Sphingobium japonicum strain UT26
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additional information
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UT26, W175 probably naturally exchanged for F or Q explaining substrate specificities
additional information
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UT26, W175 probably naturally exchanged for F or Q explaining substrate specificities
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additional information
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generation of a truncated enzyme form lacking the N-terminus, DmmAshort, from full-length wild-type enzyme, DmmAlong
additional information
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GJ10, several mutants of F172, tested for substrate binding
additional information
development and evaluation of a high-throughput system to select active haloalkane dehalogenase variants from a large mutant library, enrichment of the active wild-type enzyme in contrast to the inactive variants is about 340fold. Three saturation libraries, with a size of more than 106 cells, based on inactive variants of the haloalkane dehalogenases DhlA are successfully screened to retrieve active enzymes
additional information
engineered enzyme variants are constructed for in vivo evolution for activity toward 1-chlorohexane, more activity toward 1,6-dichlorohexane than DhlA wild-type, and increased activity toward 1,2-dibromoethane/1-bromobutane the wild-type
additional information
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GJ10, several mutants of F172, tested for substrate binding
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additional information
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engineered enzyme variants are constructed for in vivo evolution for activity toward 1-chlorohexane, more activity toward 1,6-dichlorohexane than DhlA wild-type, and increased activity toward 1,2-dibromoethane/1-bromobutane the wild-type
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