BRENDA - Enzyme Database
show all sequences of 3.1.2.29

Structural basis for the activity and substrate specificity of fluoroacetyl-CoA thioesterase FlK

Dias, M.V.; Huang, F.; Chirgadze, D.Y.; Tosin, M.; Spiteller, D.; Dry, E.F.; Leadlay, P.F.; Spencer, J.B.; Blundell, T.L.; J. Biol. Chem. 285, 22495-22504 (2010)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
His-tagged protein expressed in Escherichia coli Rosetta (DE3)pLysS
Streptomyces cattleya
Crystallization (Commentary)
Crystallization
Organism
FlK forms a dimer, in which each subunit adopts a hot dog fold as observed for type II thioesterases. Unlike other type II thioesterases, FlK employs a catalytic triad composed of Thr42, His76, and a water molecule; sitting drop method, polyethylene glycol monomethyl ether 2000
Streptomyces cattleya
Engineering
Amino acid exchange
Commentary
Organism
E50A
disturbed protein folding. 37% of wild-type catalytic efficiency, substrate inhibition above 0.019 mM; putative member of the catalytic triad
Streptomyces cattleya
H76D
insoluble protein; putative member of the catalytic triad, completely insoluble
Streptomyces cattleya
T42A
disturbed protein folding, complete loss of activity; putative member of the catalytic triad
Streptomyces cattleya
T42S
disturbed protein folding. 19fold increase in catalytic efficiency. Substrate inhibition above 0.012 mM; putative member of the catalytic triad
Streptomyces cattleya
Inhibitors
Inhibitors
Commentary
Organism
Structure
fluoroacetyl-CoA
substrate inhibition of mutants T42S and E50A above 0.012 and 0.019 mM, respectively; T42S mutant protein: substrate inhibition observed above 0.012 mM, E50A mutant protein: substrate inhibition observed above 0.019 mM
Streptomyces cattleya
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.015
-
fluoroacetyl-CoA
mutant T42S, 25°C, pH not specified in the publication; T42S mutant protein, pH not specified in the publication, 25°C
Streptomyces cattleya
0.03
-
fluoroacetyl-CoA
wild-type, 25°C, pH not specified in the publication; wild type protein, pH not specified in the publication, 25°C
Streptomyces cattleya
0.206
-
fluoroacetyl-CoA
E50A mutant protein, pH not specified in the publication, 25°C; mutant E50A, 25°C, pH not specified in the publication
Streptomyces cattleya
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Streptomyces cattleya
Q1EMV2
-
-
Purification (Commentary)
Commentary
Organism
immobilized metal ion affinity chromatography (Co2+), His tag removed, gel filtration
Streptomyces cattleya
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
fluoroacetyl-CoA + H2O
-
712448
Streptomyces cattleya
fluoroacetate + CoA
-
-
-
?
fluoroacetyl-CoA + H2O
does not accept acetyl-CoA as a substrate
712448
Streptomyces cattleya
fluoroacetate + CoA
-
-
-
?
Subunits
Subunits
Commentary
Organism
homodimer
crystal structure analysis
Streptomyces cattleya
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.044
-
fluoroacetyl-CoA
wild-type, 25°C, pH not specified in the publication; wild type protein, pH not specified in the publication, 25°C
Streptomyces cattleya
0.111
-
fluoroacetyl-CoA
E50A mutant protein, pH not specified in the publication, 25°C; mutant E50A, 25°C, pH not specified in the publication
Streptomyces cattleya
0.409
-
fluoroacetyl-CoA
mutant T42S, 25°C, pH not specified in the publication; T42S mutant protein, pH not specified in the publication, 25°C
Streptomyces cattleya
Cloned(Commentary) (protein specific)
Commentary
Organism
His-tagged protein expressed in Escherichia coli Rosetta (DE3)pLysS
Streptomyces cattleya
Crystallization (Commentary) (protein specific)
Crystallization
Organism
FlK forms a dimer, in which each subunit adopts a hot dog fold as observed for type II thioesterases. Unlike other type II thioesterases, FlK employs a catalytic triad composed of Thr42, His76, and a water molecule; sitting drop method, polyethylene glycol monomethyl ether 2000
Streptomyces cattleya
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
E50A
disturbed protein folding. 37% of wild-type catalytic efficiency, substrate inhibition above 0.019 mM; putative member of the catalytic triad
Streptomyces cattleya
H76D
insoluble protein; putative member of the catalytic triad, completely insoluble
Streptomyces cattleya
T42A
disturbed protein folding, complete loss of activity; putative member of the catalytic triad
Streptomyces cattleya
T42S
disturbed protein folding. 19fold increase in catalytic efficiency. Substrate inhibition above 0.012 mM; putative member of the catalytic triad
Streptomyces cattleya
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
fluoroacetyl-CoA
substrate inhibition of mutants T42S and E50A above 0.012 and 0.019 mM, respectively; T42S mutant protein: substrate inhibition observed above 0.012 mM, E50A mutant protein: substrate inhibition observed above 0.019 mM
Streptomyces cattleya
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.015
-
fluoroacetyl-CoA
mutant T42S, 25°C, pH not specified in the publication; T42S mutant protein, pH not specified in the publication, 25°C
Streptomyces cattleya
0.03
-
fluoroacetyl-CoA
wild-type, 25°C, pH not specified in the publication; wild type protein, pH not specified in the publication, 25°C
Streptomyces cattleya
0.206
-
fluoroacetyl-CoA
E50A mutant protein, pH not specified in the publication, 25°C; mutant E50A, 25°C, pH not specified in the publication
Streptomyces cattleya
Purification (Commentary) (protein specific)
Commentary
Organism
immobilized metal ion affinity chromatography (Co2+), His tag removed, gel filtration
Streptomyces cattleya
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
fluoroacetyl-CoA + H2O
-
712448
Streptomyces cattleya
fluoroacetate + CoA
-
-
-
?
fluoroacetyl-CoA + H2O
does not accept acetyl-CoA as a substrate
712448
Streptomyces cattleya
fluoroacetate + CoA
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
homodimer
crystal structure analysis
Streptomyces cattleya
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.044
-
fluoroacetyl-CoA
wild-type, 25°C, pH not specified in the publication; wild type protein, pH not specified in the publication, 25°C
Streptomyces cattleya
0.111
-
fluoroacetyl-CoA
E50A mutant protein, pH not specified in the publication, 25°C; mutant E50A, 25°C, pH not specified in the publication
Streptomyces cattleya
0.409
-
fluoroacetyl-CoA
mutant T42S, 25°C, pH not specified in the publication; T42S mutant protein, pH not specified in the publication, 25°C
Streptomyces cattleya
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.54
-
fluoroacetyl-CoA
E50A mutant protein, pH not specified in the publication, 25°C; mutant E50A, 25°C, pH not specified in the publication
Streptomyces cattleya
1.47
-
fluoroacetyl-CoA
wild-type, 25°C, pH not specified in the publication; wild type protein, pH not specified in the publication, 25°C
Streptomyces cattleya
27.3
-
fluoroacetyl-CoA
mutant T42S, 25°C, pH not specified in the publication; T42S mutant protein, pH not specified in the publication, 25°C
Streptomyces cattleya
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.54
-
fluoroacetyl-CoA
E50A mutant protein, pH not specified in the publication, 25°C; mutant E50A, 25°C, pH not specified in the publication
Streptomyces cattleya
1.47
-
fluoroacetyl-CoA
wild-type, 25°C, pH not specified in the publication; wild type protein, pH not specified in the publication, 25°C
Streptomyces cattleya
27.3
-
fluoroacetyl-CoA
mutant T42S, 25°C, pH not specified in the publication; T42S mutant protein, pH not specified in the publication, 25°C
Streptomyces cattleya
Other publictions for EC 3.1.2.29
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
752061
Weeks
Entropy drives selective fluo ...
Streptomyces cattleya
Proc. Natl. Acad. Sci. USA
115
E2193-E2201
2018
-
-
-
-
1
-
6
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
24
-
-
-
-
-
-
-
1
-
-
6
24
-
-
-
-
1
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
729250
Weeks
Molecular recognition of fluor ...
Streptomyces cattleya
Biochemistry
53
2053-2063
2014
-
-
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
730813
Weeks
Catalytic control of enzymatic ...
Streptomyces cattleya
Proc. Natl. Acad. Sci. USA
109
19667-19672
2012
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
711265
Weeks
Structural and biochemical stu ...
Streptomyces cattleya
Biochemistry
49
9269-9279
2010
-
-
-
1
9
-
-
15
-
-
-
-
-
1
-
-
-
-
-
-
-
-
3
-
-
-
-
13
-
-
-
-
-
-
-
-
-
-
-
1
9
-
-
-
-
15
-
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
13
-
-
-
-
-
1
1
-
15
15
712448
Dias
Structural basis for the activ ...
Streptomyces cattleya
J. Biol. Chem.
285
22495-22504
2010
-
-
1
1
4
-
1
3
-
-
-
-
-
2
-
-
1
-
-
-
-
-
2
1
-
-
-
3
-
-
-
-
-
-
-
-
-
1
-
1
4
-
-
1
-
3
-
-
-
-
-
-
-
1
-
-
-
-
2
1
-
-
-
3
-
-
-
-
-
-
-
-
3
3
712861
Meyer
-
Fluoroacetyl-coenzyme A hydrol ...
Dichapetalum cymosum
J. Plant Physiol.
139
369-372
1992
-
-
-
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
-
-
1
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
1
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-