Information on EC 3.11.1.1 - phosphonoacetaldehyde hydrolase

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The expected taxonomic range for this enzyme is: Bacteria

EC NUMBER
COMMENTARY
3.11.1.1
-
RECOMMENDED NAME
GeneOntology No.
phosphonoacetaldehyde hydrolase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
phosphonoacetaldehyde + H2O = acetaldehyde + phosphate
show the reaction diagram
Schiff base mechanism
-
phosphonoacetaldehyde + H2O = acetaldehyde + phosphate
show the reaction diagram
imine formation between the enzyme and its substrate
-
phosphonoacetaldehyde + H2O = acetaldehyde + phosphate
show the reaction diagram
double displacement mechanism proceeding via protonated Schiff base and phosphoenzyme intermediates. The mechanism involves P-C bond cleavage in a protonated Schiff base intermediate by in-line displacement by an enzyme nucleophile. Subsequent hydrolysis of the resultant acetaldehyde enamine and phosphoenzyme groups then yield acetaldehyde and phosphate
-
phosphonoacetaldehyde + H2O = acetaldehyde + phosphate
show the reaction diagram
mechanism involves Schiff base formation with Lys53 followed by phosphoryl transfer to Asp12 and at last hydrolysis at the imine and acyl phosphate phosphorus
-
phosphonoacetaldehyde + H2O = acetaldehyde + phosphate
show the reaction diagram
mechanism involves Schiff base formation with Lys53 followed by phosphoryl transfer to Asp11 and at last hydrolysis at the imine and acyl phosphate phosphorus
-
phosphonoacetaldehyde + H2O = acetaldehyde + phosphate
show the reaction diagram
Schiff base formation with catalytic Lys and phosphonoacetaldehyde, PC-bond cleavage in the Schiff base takes place during the second partial reaction and liberation of the acetaldehyde from the resulting enamine occurs during the third partial reaction
-
phosphonoacetaldehyde + H2O = acetaldehyde + phosphate
show the reaction diagram
bicovalent catalytic mechanism in which an active site nucleophile abstracts the phosphoryl group from the Schiff-base intermediate formed from Lys53 and phosphonoacetaldehyde
-
phosphonoacetaldehyde + H2O = acetaldehyde + phosphate
show the reaction diagram
mechanism, active site conformation during catalysis, Lys53 is involved
-
phosphonoacetaldehyde + H2O = acetaldehyde + phosphate
show the reaction diagram
active site structure
-
phosphonoacetaldehyde + H2O = acetaldehyde + phosphate
show the reaction diagram
mechanism, reaction pathway, Schiff base formation between an amine and a ketone in aqueous solution, active site model
-
phosphonoacetaldehyde + H2O = acetaldehyde + phosphate
show the reaction diagram
quantum chemical study of the imine formation reaction, which precedes P-C bond cleavage. The barrier of this reaction can be significantly lowered if the reaction is assisted by a water molecule and the substrate is protonated
-
phosphonoacetaldehyde + H2O = acetaldehyde + phosphate
show the reaction diagram
alternative catalytic mechanism, involving proton transfer that triggers P-C bond cleavage, transition states, TSd1, TSm1, TSd2, TSm2, and theoretical QM/MM study using crystal structure of an inhibitor-bound enzyme, overview. The bond breaking process is facilitated by proton transfer from catalytic lysine residue to the substrate. The common catalytic mechanism involves formation of a Schiff base, overview
-
phosphonoacetaldehyde + H2O = acetaldehyde + phosphate
show the reaction diagram
Schiff base formation with catalytic Lys and phosphonoacetaldehyde, PC-bond cleavage in the Schiff base takes place during the second partial reaction and liberation of the acetaldehyde from the resulting enamine occurs during the third partial reaction
Bacillus cereus AI-2
-
-
phosphonoacetaldehyde + H2O = acetaldehyde + phosphate
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of carbon-phosphorus bond
-
-
-
-
P-C bond cleavage
O31156
-
PATHWAY
KEGG Link
MetaCyc Link
2-aminoethylphosphonate degradation I
-
Metabolic pathways
-
Microbial metabolism in diverse environments
-
Phosphonate and phosphinate metabolism
-
SYSTEMATIC NAME
IUBMB Comments
2-oxoethylphosphonate phosphonohydrolase
This enzyme destabilizes the C-P bond, by forming an imine between one of its lysine residues and the carbonyl group of the substrate, thus allowing this, normally stable, bond to be broken. The mechanism is similar to that used by EC 4.1.2.13, fructose-bisphosphate aldolase, to break a C-C bond. Belongs to the haloacetate dehalogenase family.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2-phosphonoacetaldehyde phosphonohydrolase
-
-
2-phosphonoacetylaldehyde phosphonohydrolase
-
-
-
-
hydrolase, phosphonoacetylaldehyde
-
-
-
-
P-Ald hydrolase
-
-
-
-
phosphonatase
-
-
-
-
phosphonatase
-
-
CAS REGISTRY NUMBER
COMMENTARY
37289-42-2
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Bacillus cereus AI-2
AI-2
-
-
Manually annotated by BRENDA team
strain IFO 12010. 2-aminoethylphosphonic acid:pyruvate aminotransferase and phosphonoacetaldehyde hydrolase activities are induced when cells are both phosphate limited and supplied with 2-aminoethylphosphonic acid as sole source of phosphorus in the culture medium. Neither enzyme is induced in phosphate-replete medium, or in medium where both 2-aminoethylphosphoonic acid and phosphate are supplied
-
-
Manually annotated by BRENDA team
Enterobacter aerogenes IFO 12010
strain IFO 12010. 2-aminoethylphosphonic acid:pyruvate aminotransferase and phosphonoacetaldehyde hydrolase activities are induced when cells are both phosphate limited and supplied with 2-aminoethylphosphonic acid as sole source of phosphorus in the culture medium. Neither enzyme is induced in phosphate-replete medium, or in medium where both 2-aminoethylphosphoonic acid and phosphate are supplied
-
-
Manually annotated by BRENDA team
strain A237
-
-
Manually annotated by BRENDA team
Pseudomonas aeruginosa A237
A237
-
-
Manually annotated by BRENDA team
Pseudomonas aeruginosa A237
strain A237
-
-
Manually annotated by BRENDA team
strain NG2, can utilize 2-aminoethylphosphonic acid as sole carbon and energy, nitrogen and phosphorus source. Both 2-aminoethylphosphonic acid:pyruvate aminotransferase and phosphonoacetaldehyde hydrolase activities are inducible by the presence of 2-aminoethylphosphonic acid in the culture medium
-
-
Manually annotated by BRENDA team
Pseudomonas putida NG2
strain NG2, can utilize 2-aminoethylphosphonic acid as sole carbon and energy, nitrogen and phosphorus source. Both 2-aminoethylphosphonic acid:pyruvate aminotransferase and phosphonoacetaldehyde hydrolase activities are inducible by the presence of 2-aminoethylphosphonic acid in the culture medium
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
metabolism
-
the enzyme is involved in the phosphonatase pathway of 2-aminoethylphosphonic acid degradation, overview
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetonyl phosphonate + H2O
?
show the reaction diagram
Bacillus cereus, Bacillus cereus AI-2
-
-
-
-
-
p-nitrophenylphosphate + H2O
p-nitrophenol + phosphate
show the reaction diagram
-
hydrolyzed at considerable lower rate than phosphonoacetaldehyde
-
-
?
phosphonoacetaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
-
-
-
?
phosphonoacetaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
-
-
ir
phosphonoacetaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
-
-
-
?
phosphonoacetaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
i.e. Pald
-
?
phosphonoacetaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
O31156
i.e. Pald, cleavage via Schiff base intermediate formed with Lys53, bound substrate stabilizes the closed conformation of the active site, thus facilitating catalysis
-
?
phosphonoacetaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
catalysis within the core domain of phosphonatase requires the participation of loop 5 of the corresponding cap domain. Gly is an indispensable component
-
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
-
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
-
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
-
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
-
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
-
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
-
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
-
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
-
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
2-phosphonoacetaldehyde
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
second step in the pathway by which Bacillus cereus metabolizes 2-aminoethylphosphonic acid
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
second step in the pathway by which Bacillus cereus metabolizes 2-aminoethylphosphonic acid
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
second step in the pathway by which Bacillus cereus metabolizes 2-aminoethylphosphonic acid
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
Bacillus cereus AI-2
-
-
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
Pseudomonas aeruginosa A237
-
-
-
?
thiophosphonoacetaldehyde + H2O
thiophosphate + acetaldehyde
show the reaction diagram
Bacillus cereus, Bacillus cereus AI-2
-
-
-
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
phosphonoacetaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
-
-
-
?
phosphonoacetaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
-
-
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
-
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
-
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
second step in the pathway by which Bacillus cereus metabolizes 2-aminoethylphosphonic acid
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
second step in the pathway by which Bacillus cereus metabolizes 2-aminoethylphosphonic acid
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
-
second step in the pathway by which Bacillus cereus metabolizes 2-aminoethylphosphonic acid
-
?
phosphonoacetylaldehyde + H2O
acetaldehyde + phosphate
show the reaction diagram
Pseudomonas aeruginosa A237
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
copper
-
contains 0.3 gatom per molecular weight of 75000 Da
Iron
-
contains trace amounts of iron
Mg2+
-
required; stabilizes the active homodimer form of the enzyme
Mg2+
-
required
Mg2+
-
cofactor
Mg2+
-
activates, required for catalysis, serves as a cofactor, binds via ligation to the loop 1 Asp12 carboxylate and Thr 14 backbone carbonyl and to the loop 4 Asp186carboxylate, the loop 4 Asp190 forms a hydrogen bond to the Mg(II) water ligand, Asp186 is essential while Asp190 simply enhances cofactor binding
Mg2+
-
required for catalysis
Zinc
-
contains 0.4 gatom per molecular weight of 75000 Da
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2,4-Dinitrophenylacetate
-
loss of activity due to acetylation of Ls53 with the inert compound, pH profile of inactivation
Acetonylphosphonate
-
-
Acetonylphosphonate
-
in presence of NH4+
fluorophosphate
-
competitive inhibitor
Malonic semialdehyde
-
competitive inhibitor
n-butylphosphonic acid
-
activates at low concentrations, inhibits at high concentrations. Allosteric model involving two different classes of sites for n-butylphosphonic acid
NaBH4
-
in presence of phosphonoacetaldehyde or acetaldehyde; no effect in absence of substrate or product
NaBH4
-
in presence of phosphonoacetaldehyde or acetaldehyde
phosphonoacetaldehyde
-
competitive inhibitor
phosphonoethanol
-
competitive inhibitor
Trypsin
-
degradation
-
vinyl sulfonate
-
competitive, inhibition mechanism
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-Aminoethylphosphonic acid
-
both 2-aminoethylphosphonic acid:pyruvate aminotransferase and phosphonoacetaldehyde hydrolase activities are inducible by the presence of 2-aminoethylphosphonic acid in the culture medium
n-butylphosphonic acid
-
activates at low concentrations, inhibits at high concentrations. Allosteric model involving two different classes of sites for n-butylphosphonic acid
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.033
-
phosphonoacetaldehyde
-
wild-type enzyme, pH 7.5, 25C
0.033
-
phosphonoacetaldehyde
-
wild-type enzyme, mutant C22S, and mutant Y128F/C22S, pH 7.5, 25C
0.033
-
phosphonoacetaldehyde
-
25C, pH 7.5, wild-type enzyme
0.035
-
phosphonoacetaldehyde
-
mutant Y128A, pH 7.5, 25C
0.04
-
phosphonoacetaldehyde
-
-
0.045
-
phosphonoacetaldehyde
-
mutant Y128F, pH 7.5, 25C
0.054
-
phosphonoacetaldehyde
-
mutant G185D/D190G, pH 7.5, 25C
0.056
-
phosphonoacetaldehyde
-
triple mutant K121R/K146R/K192R, pH 7.5, 25C
0.072
-
phosphonoacetaldehyde
-
mutant D12E, pH 7.5, 25C
0.145
-
phosphonoacetaldehyde
-
mutant H56A, pH 7.5, 25C
0.175
-
phosphonoacetaldehyde
-
25C, pH 7.5, mutant enzyme H56Q
0.193
-
phosphonoacetaldehyde
-
mutant K183A, pH 7.5, 25C
0.52
-
phosphonoacetaldehyde
-
mutant D190A, pH 7.5, 25C
0.53
-
phosphonoacetaldehyde
-
mutant C22A, pH 7.5, 25C
0.77
-
phosphonoacetaldehyde
-
mutant K183L, pH 7.5, 25C
5.1
-
phosphonoacetaldehyde
-
mutant M49L, pH 7.5, 25C
5.1
-
phosphonoacetaldehyde
-
25C, pH 7.5, mutant enzyme m49L
11
-
phosphonoacetaldehyde
-
25C, pH 7.5, mutant enzyme G50A
additional information
-
additional information
-
kinetics
-
additional information
-
additional information
-
kinetics, activity of mutant D186A is too low to measure Km accurately
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
16.7
-
2-phosphonoacetaldehyde
-
-
8.4e-05
-
phosphonoacetaldehyde
-
mutant D186A, pH 7.5, 25C
0.0012
-
phosphonoacetaldehyde
-
mutant D12E, pH 7.5, 25C
0.00247
-
phosphonoacetaldehyde
-
25C, pH 7.5, mutant enzyme m49L
0.006
-
phosphonoacetaldehyde
-
25C, pH 7.5, mutant enzyme H56Q
0.0071
-
phosphonoacetaldehyde
-
25C, pH 7.5, mutant enzyme G50A
0.012
-
phosphonoacetaldehyde
-
mutant K183A, pH 7.5, 25C
0.022
-
phosphonoacetaldehyde
-
mutant D190A, pH 7.5, 25C
0.046
-
phosphonoacetaldehyde
-
mutant K183L, pH 7.5, 25C
0.075
-
phosphonoacetaldehyde
-
mutant H56A, pH 7.5, 25C
0.077
-
phosphonoacetaldehyde
-
mutant Y128A, pH 7.5, 25C
0.25
-
phosphonoacetaldehyde
-
25C, pH 7.5, wild-type enzyme
1.28
-
phosphonoacetaldehyde
-
triple mutant K121R/K146R/K192R, pH 7.5, 25C
1.7
-
phosphonoacetaldehyde
-
mutant G185D/D190G, pH 7.5, 25C
1.95
-
phosphonoacetaldehyde
-
mutant C22A, pH 7.5, 25C
2.11
-
phosphonoacetaldehyde
-
mutant Y128F/C22S, pH 7.5, 25C
2.21
-
phosphonoacetaldehyde
-
mutant Y128F, pH 7.5, 25C
2.26
-
phosphonoacetaldehyde
-
mutant C22S, pH 7.5, 25C
2.94
-
phosphonoacetaldehyde
-
mutant C22S, pH 7.5, 25C; mutant Y128F/C22S, pH 7.5, 25C; mutant Y128F, pH 7.5, 25C
6.08
-
phosphonoacetaldehyde
-
triple mutant K121R/K146R/K192R, pH 7.5, 25C
15
-
phosphonoacetaldehyde
-
wild-type enzyme, pH 7.5, 25C
15
-
phosphonoacetaldehyde
-
wild-type enzyme and mutant M49L, pH 7.5, 25C
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1.79
-
vinyl sulfonate
-
pH 7.5, 25C
additional information
-
additional information
-
inhibition kinetics
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
13.9
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.5
-
-
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.5
10
-
pH 6.5: about 35% of maximal activity, pH 10.0: about 30% of maximal activity
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
assy at
25
-
-
assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
55
-
30C: about 50% of maximal activity, 55C: about 45% of maximal activity
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
68000
-
-
nondenaturing PAGE
83000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
dimer
-
2 * 33000-37000, similar subunits, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
10 mg/ml purified recombinant wild-type and mutant enzymes, complexed with Mg2+ only or with Mg2+ and inhibitor vinyl sulfonate, in 1 mM HEPES, 10 mM MgCl2, 0.1 mM DTT, pH 7.5, 4C, hanging drop vapour diffusion method, equal volume of protein and reservoir solution, the latter containing 30% PEG 4000, 100 mM Tris-HCl, pH 7.4, 100 mM MgCl2, 1 week, against the reservoir well solution additionally with 20% glycerol before data collection, X-ray diffraction structure determination and analysis at 2.4-2.8 A resolution
-
10 mg/ml wild-type and mutant D12A enzymes complexed with Mg2+ only or with Mg2+ and substrate, in 1 mM HEPES, 10 mM MgCl2, 0.1 mM DTT, pH 7.5, 4C, hanging drop vapour diffusion method, equal volume of protein and reservoir solution, the latter containing 30% PEG 4000, 100 mM Tris-HCl, pH 7.4, 100 mM MgCl2, 1 week, against the reservoir well solution additionally with 20% glycerol before data collection, X-ray diffraction structure determination and analysis at 2.3-2.55 A
-
crystal structure of the homodimeric enzyme complexed with the phosphate analogue tungstate and Mg2+
-
mutant enzymes
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6
-
-
protonation of the active site Lys is the cause for loss of activity
8
-
-
deprotonation of the active site Cys may be the cause for the loss of enzyme activity
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
35
-
-
rapid loss loss of activity in absence of Mg2+
45
-
-
heat-labile, even in presence of 5 mM Mg2+
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
inactivation in absence of Mg2+ is aggravated at higher pH or when either EDTA or Ca2+ is added
-
repeated freezing and thawing causes rapid loss of activity
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, stable for several months
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant wild-type and mutant enzymes from Escherichia coli
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression of wild-type and mutant enzymes in Escherichia coli
-
expression of wild-type and mutant enzymes in Escherichia coli JM109
-
expression in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C22A
-
site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme
C22S
-
site-directed mutagenesis, reduced activity compared to the wild-type enzyme
D12A
-
site-directed mutagenesis, catalytically inactive mutant
D186A
-
site-directed mutagenesis, highly reduced activity
D186A/D190A
-
site-directed mutagenesis, inactive mutant
D186E
-
site-directed mutagenesis, very highly reduced activity
D190A
-
site-directed mutagenesis, reduced activity
G185D/D190G
-
site-directed mutagenesis, reduced activity
G50A
-
kcat/Km is 12820fold lower than wild-type value
G50P
-
inactive mutant protein
G50V
-
inactive mutant protein
H56A
-
site-directed mutagenesis, very highly reduced activity compared to the wild-type enzyme
H56Q
-
kcat/Km is 238fold lower than wild-type value
K121R/K146R/K192R
-
site-directed mutagenesis, reduced activity compared to the wild-type enzyme
K183A
-
site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme, Lys183 is probably important in maintaining the active site environment
K183L
-
site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme, Lys183 is probably important in maintaining the active site environment
K53A
-
inactive mutant protein
K53R
-
inactive mutant protein
M49L
-
site-directed mutagenesis, very highly reduced activity compared to the wild-type enzyme
M49L
-
kcat/Km is 17241fold lower than wild-type value
Y128A
-
site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme
Y128F
-
site-directed mutagenesis, reduced activity compared to the wild-type enzyme
Y128F/C22S
-
site-directed mutagenesis, reduced activity compared to the wild-type enzyme
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression of mutant enzymes in escherichia coli
-