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show all sequences of 3.1.1.41

Structural role of a conserved active site cis proline in the Thermotoga maritima acetyl esterase from the carbohydrate esterase family 7

Singh, M.; Manoj, N.; Proteins 85, 694-708 (2017)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli DH5alpha cells
Thermotoga maritima
Crystallization (Commentary)
Crystallization
Organism
crystal structure of the P228A variant is determined at 2.1 A resolution. The replacement does not affect the overall secondary, tertiary, and quaternary structures and moderately decreases the thermal stability; mutant enzyme P228A, hanging drop vapor diffusion method, using 0.05M ammonium sulfate, 0.1 M Bis-Tris pH 6.5 and 30% (v/v) pentaerythritolethoxylate
Thermotoga maritima
Engineering
Amino acid exchange
Commentary
Organism
additional information
construction of truncation variants lacking four and ten residues of the N-terminal region. The truncations do not affect the secondary structure or the fold but modulate the oligomerization dynamics. A modest in substrate specificity for acetylated xylose is observed compared with acetylated glucose. Variants show a drastic reduction of 30-40°C in the optimum temperature for activity
Thermotoga maritima
P228A
the mutation does not affect the overall secondary, tertiary, and quaternary structures and moderately decreases the thermal stability. However, the wild type cis conformation of the 227-228 peptide bond adopts a trans conformation in the variant. The results suggest that the conserved proline residue is responsible for the cis conformation of the peptide and shapes the geometry of the active site. Elimination of the pyrrolidine ring results in the loss of van der Waals and hydrophobic interactions with both the alcohol and acyl moeities of the ester substrate, leading to significant impairment of the activity and perturbation of substrate specificity; the mutation does not affect the overall secondary, tertiary, and quaternary structures, but has a significantly decreasing effect on the activity and moderately decreases the thermal stability of the enzyme. The wild type cis conformation of the 227-228 peptide bond adopts a trans conformation in the variant
Thermotoga maritima
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.059
-
xylose tetraacetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
0.134
-
xylose tetraacetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C; wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
0.148
-
4-methylumbelliferyl acetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
0.175
-
4-methylumbelliferyl acetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
0.175
-
4-nitrophenyl acetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
0.187
-
4-methylumbelliferyl acetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
0.189
-
xylose tetraacetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
0.193
-
4-nitrophenyl acetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
0.197
-
4-methylumbelliferyl acetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
0.203
-
Glucose pentaacetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
0.207
-
Glucose pentaacetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C; wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
0.224
-
Glucose pentaacetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
0.231
-
4-nitrophenyl acetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
0.309
-
4-nitrophenyl acetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
1.12
-
7-aminocephalosporanic acid
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
1.135
-
7-aminocephalosporanic acid
wild-type, pH 8.0, 50°C
Thermotoga maritima
1.384
-
7-aminocephalosporanic acid
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
1.563
-
7-aminocephalosporanic acid
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
76000
78000
sedimentation equilibrium centrifugation, minor peak
Thermotoga maritima
210000
220000
sedimentation equilibrium centrifugation, dominant peak
Thermotoga maritima
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Thermotoga maritima
Q9WXT2
-
-
Thermotoga maritima ATCC 43589
Q9WXT2
-
-
Thermotoga maritima DSM 3109
Q9WXT2
-
-
Purification (Commentary)
Commentary
Organism
-
Thermotoga maritima
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.075
-
substrate: 7-aminocephalosporanic acid, pH 8.0, 50°C, mutant enzyme P228A; with 7-amino-cephalosporanic acid as substrate, mutant enzyme P228A, at pH 8.0 and 50°C
Thermotoga maritima
0.47
-
substrate: 1-naphthyl acetate, pH 8.0, 50°C, mutant enzyme P228A; with 1-naphthyl acetate as substrate, mutant enzyme P228A, at pH 8.0 and 50°C
Thermotoga maritima
1.78
-
substrate: 4-nitrophenyl acetate, pH 8.0, 50°C, mutant enzyme P228A; with 4-nitrophenyl acetate as substrate, mutant enzyme P228A, at pH 8.0 and 50°C
Thermotoga maritima
3.98
-
substrate: 4-methylumbelliferyl acetate, pH 8.0, 50°C, mutant enzyme P228A; with 4-methylumbelliferyl acetate as substrate, mutant enzyme P228A, at pH 8.0 and 50°C
Thermotoga maritima
10
-
substrate: 7-aminocephalosporanic acid, pH 8.0, 50°C, wild-type enzyme; with 7-amino-cephalosporanic acid as substrate, wild type enzyme, at pH 8.0 and 50°C
Thermotoga maritima
16.81
-
substrate: 1-naphthyl acetate, pH 8.0, 50°C, wild-type enzyme
Thermotoga maritima
16.91
-
with 1-naphthyl acetate as substrate, wild type enzyme, at pH 8.0 and 50°C
Thermotoga maritima
25.31
-
substrate: D-glucose pentaacetate, pH 8.0, 50°C, mutant enzyme P228A; with D-glucose pentaacetate as substrate, mutant enzyme P228A, at pH 8.0 and 50°C
Thermotoga maritima
29.8
-
substrate: D-xylose tetraacetate, pH 8.0, 50°C, mutant enzyme P228A; with D-xylose tetraacetate as substrate, mutant enzyme P228A, at pH 8.0 and 50°C
Thermotoga maritima
41.49
-
substrate: 4-methylumbelliferyl acetate, pH 8.0, 50°C, wild-type enzyme; with 4-methylumbelliferyl acetate as substrate, wild type enzyme, at pH 8.0 and 50°C
Thermotoga maritima
49.33
-
substrate: 4-nitrophenyl acetate, pH 8.0, 50°C, wild-type enzyme; with 4-nitrophenyl acetate as substrate, wild type enzyme, at pH 8.0 and 50°C
Thermotoga maritima
346.22
-
substrate: D-xylose tetraacetate, pH 8.0, 50°C, wild-type enzyme; with D-xylose tetraacetate as substrate, wild type enzyme, at pH 8.0 and 50°C
Thermotoga maritima
415.5
-
substrate: D-glucose pentaacetate, pH 8.0, 50°C, wild-type enzyme; with D-glucose pentaacetate as substrate, wild type enzyme, at pH 8.0 and 50°C
Thermotoga maritima
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1-naphthyl acetate + H2O
-
749261
Thermotoga maritima
1-naphthol + acetate
-
-
-
?
1-naphthyl acetate + H2O
-
749261
Thermotoga maritima DSM 3109
1-naphthol + acetate
-
-
-
?
1-naphthyl acetate + H2O
-
749261
Thermotoga maritima ATCC 43589
1-naphthol + acetate
-
-
-
?
4-methylumbelliferyl acetate + H2O
-
749261
Thermotoga maritima
4-methylumbelliferol + acetate
-
-
-
?
4-methylumbelliferyl acetate + H2O
-
749261
Thermotoga maritima DSM 3109
4-methylumbelliferol + acetate
-
-
-
?
4-methylumbelliferyl acetate + H2O
-
749261
Thermotoga maritima ATCC 43589
4-methylumbelliferol + acetate
-
-
-
?
4-nitrophenyl acetate + H2O
-
749261
Thermotoga maritima
4-nitrophenol + acetate
-
-
-
?
4-nitrophenyl acetate + H2O
-
749261
Thermotoga maritima DSM 3109
4-nitrophenol + acetate
-
-
-
?
4-nitrophenyl acetate + H2O
-
749261
Thermotoga maritima ATCC 43589
4-nitrophenol + acetate
-
-
-
?
7-amino-cephalosporanic acid + H2O
-
749261
Thermotoga maritima
7-amino-3-deacetylcephalosporanic acid + acetate
-
-
-
?
7-aminocephalosporanic acid + H2O
-
749261
Thermotoga maritima
7-amino-3-deacetylcephalosporanic acid + acetate
-
-
-
?
7-aminocephalosporanic acid + H2O
-
749261
Thermotoga maritima
acetate + H2O
-
-
-
?
D-glucose pentaacetate + H2O
-
749261
Thermotoga maritima
D-glucose + acetate
-
-
-
?
D-glucose pentaacetate + H2O
-
749261
Thermotoga maritima DSM 3109
D-glucose + acetate
-
-
-
?
D-glucose pentaacetate + H2O
-
749261
Thermotoga maritima ATCC 43589
D-glucose + acetate
-
-
-
?
D-glucose pentaacetate + H2O
-
749261
Thermotoga maritima
?
-
-
-
?
D-glucose pentaacetate + H2O
-
749261
Thermotoga maritima DSM 3109
?
-
-
-
?
D-glucose pentaacetate + H2O
-
749261
Thermotoga maritima ATCC 43589
?
-
-
-
?
D-xylose tetraacetate + H2O
-
749261
Thermotoga maritima
?
-
-
-
?
D-xylose tetraacetate + H2O
-
749261
Thermotoga maritima
D-xylose + acetate
-
-
-
?
fluorescein di(acetoxymethyl ether) + H2O
-
749261
Thermotoga maritima
?
-
-
-
?
glucose pentaacetate + H2O
-
749261
Thermotoga maritima
? + acetate
-
-
-
?
xylose tetraacetate + H2O
-
749261
Thermotoga maritima
? + acetate
-
-
-
?
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
50
-
assay at
Thermotoga maritima
60
70
truncation variants lacking four and ten residues of the N-terminus
Thermotoga maritima
95
-
excessive auto-hydrolysis of the substrate at higher temperatures limits the assay at 95°C
Thermotoga maritima
Temperature Stability [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
105
-
the unfolding transition for the wild type occurs at a temperature higher than 105°C
Thermotoga maritima
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
15.5
-
7-aminocephalosporanic acid
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
20.2
-
7-aminocephalosporanic acid
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
23.8
-
7-aminocephalosporanic acid
wild-type, pH 8.0, 50°C
Thermotoga maritima
24.8
-
7-aminocephalosporanic acid
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
32.2
-
4-methylumbelliferyl acetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
39.8
-
4-methylumbelliferyl acetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
42.7
-
4-methylumbelliferyl acetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
47.8
-
4-nitrophenyl acetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
48.7
-
4-nitrophenyl acetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
60.3
-
4-nitrophenyl acetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
63
-
4-methylumbelliferyl acetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
96.2
-
4-nitrophenyl acetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
169.8
-
xylose tetraacetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
266.7
-
xylose tetraacetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
280.7
-
xylose tetraacetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
293.2
-
Glucose pentaacetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
295.5
-
Glucose pentaacetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
323.2
-
Glucose pentaacetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
326
-
xylose tetraacetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
347
-
Glucose pentaacetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
; assay at
Thermotoga maritima
pH Range
pH Minimum
pH Maximum
Commentary
Organism
7.5
8.5
-
Thermotoga maritima
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli DH5alpha cells
Thermotoga maritima
Crystallization (Commentary) (protein specific)
Crystallization
Organism
crystal structure of the P228A variant is determined at 2.1 A resolution. The replacement does not affect the overall secondary, tertiary, and quaternary structures and moderately decreases the thermal stability; mutant enzyme P228A, hanging drop vapor diffusion method, using 0.05M ammonium sulfate, 0.1 M Bis-Tris pH 6.5 and 30% (v/v) pentaerythritolethoxylate
Thermotoga maritima
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
additional information
construction of truncation variants lacking four and ten residues of the N-terminal region. The truncations do not affect the secondary structure or the fold but modulate the oligomerization dynamics. A modest in substrate specificity for acetylated xylose is observed compared with acetylated glucose. Variants show a drastic reduction of 30-40°C in the optimum temperature for activity
Thermotoga maritima
P228A
the mutation does not affect the overall secondary, tertiary, and quaternary structures and moderately decreases the thermal stability. However, the wild type cis conformation of the 227-228 peptide bond adopts a trans conformation in the variant. The results suggest that the conserved proline residue is responsible for the cis conformation of the peptide and shapes the geometry of the active site. Elimination of the pyrrolidine ring results in the loss of van der Waals and hydrophobic interactions with both the alcohol and acyl moeities of the ester substrate, leading to significant impairment of the activity and perturbation of substrate specificity; the mutation does not affect the overall secondary, tertiary, and quaternary structures, but has a significantly decreasing effect on the activity and moderately decreases the thermal stability of the enzyme. The wild type cis conformation of the 227-228 peptide bond adopts a trans conformation in the variant
Thermotoga maritima
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.059
-
xylose tetraacetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
0.134
-
xylose tetraacetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C; wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
0.148
-
4-methylumbelliferyl acetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
0.175
-
4-methylumbelliferyl acetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
0.175
-
4-nitrophenyl acetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
0.187
-
4-methylumbelliferyl acetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
0.189
-
xylose tetraacetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
0.193
-
4-nitrophenyl acetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
0.197
-
4-methylumbelliferyl acetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
0.203
-
Glucose pentaacetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
0.207
-
Glucose pentaacetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C; wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
0.224
-
Glucose pentaacetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
0.231
-
4-nitrophenyl acetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
0.309
-
4-nitrophenyl acetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
1.12
-
7-aminocephalosporanic acid
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
1.135
-
7-aminocephalosporanic acid
wild-type, pH 8.0, 50°C
Thermotoga maritima
1.384
-
7-aminocephalosporanic acid
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
1.563
-
7-aminocephalosporanic acid
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
76000
78000
sedimentation equilibrium centrifugation, minor peak
Thermotoga maritima
210000
220000
sedimentation equilibrium centrifugation, dominant peak
Thermotoga maritima
Purification (Commentary) (protein specific)
Commentary
Organism
-
Thermotoga maritima
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.075
-
substrate: 7-aminocephalosporanic acid, pH 8.0, 50°C, mutant enzyme P228A; with 7-amino-cephalosporanic acid as substrate, mutant enzyme P228A, at pH 8.0 and 50°C
Thermotoga maritima
0.47
-
substrate: 1-naphthyl acetate, pH 8.0, 50°C, mutant enzyme P228A; with 1-naphthyl acetate as substrate, mutant enzyme P228A, at pH 8.0 and 50°C
Thermotoga maritima
1.78
-
substrate: 4-nitrophenyl acetate, pH 8.0, 50°C, mutant enzyme P228A; with 4-nitrophenyl acetate as substrate, mutant enzyme P228A, at pH 8.0 and 50°C
Thermotoga maritima
3.98
-
substrate: 4-methylumbelliferyl acetate, pH 8.0, 50°C, mutant enzyme P228A; with 4-methylumbelliferyl acetate as substrate, mutant enzyme P228A, at pH 8.0 and 50°C
Thermotoga maritima
10
-
substrate: 7-aminocephalosporanic acid, pH 8.0, 50°C, wild-type enzyme; with 7-amino-cephalosporanic acid as substrate, wild type enzyme, at pH 8.0 and 50°C
Thermotoga maritima
16.81
-
substrate: 1-naphthyl acetate, pH 8.0, 50°C, wild-type enzyme
Thermotoga maritima
16.91
-
with 1-naphthyl acetate as substrate, wild type enzyme, at pH 8.0 and 50°C
Thermotoga maritima
25.31
-
substrate: D-glucose pentaacetate, pH 8.0, 50°C, mutant enzyme P228A; with D-glucose pentaacetate as substrate, mutant enzyme P228A, at pH 8.0 and 50°C
Thermotoga maritima
29.8
-
substrate: D-xylose tetraacetate, pH 8.0, 50°C, mutant enzyme P228A; with D-xylose tetraacetate as substrate, mutant enzyme P228A, at pH 8.0 and 50°C
Thermotoga maritima
41.49
-
substrate: 4-methylumbelliferyl acetate, pH 8.0, 50°C, wild-type enzyme; with 4-methylumbelliferyl acetate as substrate, wild type enzyme, at pH 8.0 and 50°C
Thermotoga maritima
49.33
-
substrate: 4-nitrophenyl acetate, pH 8.0, 50°C, wild-type enzyme; with 4-nitrophenyl acetate as substrate, wild type enzyme, at pH 8.0 and 50°C
Thermotoga maritima
346.22
-
substrate: D-xylose tetraacetate, pH 8.0, 50°C, wild-type enzyme; with D-xylose tetraacetate as substrate, wild type enzyme, at pH 8.0 and 50°C
Thermotoga maritima
415.5
-
substrate: D-glucose pentaacetate, pH 8.0, 50°C, wild-type enzyme; with D-glucose pentaacetate as substrate, wild type enzyme, at pH 8.0 and 50°C
Thermotoga maritima
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1-naphthyl acetate + H2O
-
749261
Thermotoga maritima
1-naphthol + acetate
-
-
-
?
1-naphthyl acetate + H2O
-
749261
Thermotoga maritima DSM 3109
1-naphthol + acetate
-
-
-
?
1-naphthyl acetate + H2O
-
749261
Thermotoga maritima ATCC 43589
1-naphthol + acetate
-
-
-
?
4-methylumbelliferyl acetate + H2O
-
749261
Thermotoga maritima
4-methylumbelliferol + acetate
-
-
-
?
4-methylumbelliferyl acetate + H2O
-
749261
Thermotoga maritima DSM 3109
4-methylumbelliferol + acetate
-
-
-
?
4-methylumbelliferyl acetate + H2O
-
749261
Thermotoga maritima ATCC 43589
4-methylumbelliferol + acetate
-
-
-
?
4-nitrophenyl acetate + H2O
-
749261
Thermotoga maritima
4-nitrophenol + acetate
-
-
-
?
4-nitrophenyl acetate + H2O
-
749261
Thermotoga maritima DSM 3109
4-nitrophenol + acetate
-
-
-
?
4-nitrophenyl acetate + H2O
-
749261
Thermotoga maritima ATCC 43589
4-nitrophenol + acetate
-
-
-
?
7-amino-cephalosporanic acid + H2O
-
749261
Thermotoga maritima
7-amino-3-deacetylcephalosporanic acid + acetate
-
-
-
?
7-aminocephalosporanic acid + H2O
-
749261
Thermotoga maritima
7-amino-3-deacetylcephalosporanic acid + acetate
-
-
-
?
7-aminocephalosporanic acid + H2O
-
749261
Thermotoga maritima
acetate + H2O
-
-
-
?
D-glucose pentaacetate + H2O
-
749261
Thermotoga maritima
D-glucose + acetate
-
-
-
?
D-glucose pentaacetate + H2O
-
749261
Thermotoga maritima DSM 3109
D-glucose + acetate
-
-
-
?
D-glucose pentaacetate + H2O
-
749261
Thermotoga maritima ATCC 43589
D-glucose + acetate
-
-
-
?
D-glucose pentaacetate + H2O
-
749261
Thermotoga maritima
?
-
-
-
?
D-glucose pentaacetate + H2O
-
749261
Thermotoga maritima DSM 3109
?
-
-
-
?
D-glucose pentaacetate + H2O
-
749261
Thermotoga maritima ATCC 43589
?
-
-
-
?
D-xylose tetraacetate + H2O
-
749261
Thermotoga maritima
?
-
-
-
?
D-xylose tetraacetate + H2O
-
749261
Thermotoga maritima
D-xylose + acetate
-
-
-
?
fluorescein di(acetoxymethyl ether) + H2O
-
749261
Thermotoga maritima
?
-
-
-
?
glucose pentaacetate + H2O
-
749261
Thermotoga maritima
? + acetate
-
-
-
?
xylose tetraacetate + H2O
-
749261
Thermotoga maritima
? + acetate
-
-
-
?
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
50
-
assay at
Thermotoga maritima
60
70
truncation variants lacking four and ten residues of the N-terminus
Thermotoga maritima
95
-
excessive auto-hydrolysis of the substrate at higher temperatures limits the assay at 95°C
Thermotoga maritima
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
105
-
the unfolding transition for the wild type occurs at a temperature higher than 105°C
Thermotoga maritima
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
15.5
-
7-aminocephalosporanic acid
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
20.2
-
7-aminocephalosporanic acid
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
23.8
-
7-aminocephalosporanic acid
wild-type, pH 8.0, 50°C
Thermotoga maritima
24.8
-
7-aminocephalosporanic acid
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
32.2
-
4-methylumbelliferyl acetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
39.8
-
4-methylumbelliferyl acetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
42.7
-
4-methylumbelliferyl acetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
47.8
-
4-nitrophenyl acetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
48.7
-
4-nitrophenyl acetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
60.3
-
4-nitrophenyl acetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
63
-
4-methylumbelliferyl acetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
96.2
-
4-nitrophenyl acetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
169.8
-
xylose tetraacetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
266.7
-
xylose tetraacetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
280.7
-
xylose tetraacetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
293.2
-
Glucose pentaacetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
295.5
-
Glucose pentaacetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
323.2
-
Glucose pentaacetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
326
-
xylose tetraacetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
347
-
Glucose pentaacetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
; assay at
Thermotoga maritima
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
7.5
8.5
-
Thermotoga maritima
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
9
-
7-aminocephalosporanic acid
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
18
-
7-aminocephalosporanic acid
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C; wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
21
-
7-aminocephalosporanic acid
wild-type, pH 8.0, 50°C
Thermotoga maritima
155
-
4-nitrophenyl acetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
171
-
4-methylumbelliferyl acetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
211
-
4-nitrophenyl acetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
228
-
4-methylumbelliferyl acetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
289
-
4-methylumbelliferyl acetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
320
-
4-methylumbelliferyl acetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
344
-
4-nitrophenyl acetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
497
-
4-nitrophenyl acetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
1418
-
Glucose pentaacetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
1456
-
Glucose pentaacetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
1555
-
Glucose pentaacetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
1558
-
Glucose pentaacetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
1725
-
xylose tetraacetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
1994
-
xylose tetraacetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
2083
-
xylose tetraacetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
2866
-
xylose tetraacetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
9
-
7-aminocephalosporanic acid
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
18
-
7-aminocephalosporanic acid
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C; wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
21
-
7-aminocephalosporanic acid
wild-type, pH 8.0, 50°C
Thermotoga maritima
155
-
4-nitrophenyl acetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
171
-
4-methylumbelliferyl acetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
211
-
4-nitrophenyl acetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
228
-
4-methylumbelliferyl acetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
289
-
4-methylumbelliferyl acetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
320
-
4-methylumbelliferyl acetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
344
-
4-nitrophenyl acetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
497
-
4-nitrophenyl acetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
1418
-
Glucose pentaacetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
1456
-
Glucose pentaacetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
1555
-
Glucose pentaacetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
1558
-
Glucose pentaacetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
1725
-
xylose tetraacetate
wild-type, pH 8.0, 50°C
Thermotoga maritima
1994
-
xylose tetraacetate
mutant lacking 4 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
2083
-
xylose tetraacetate
wild-type including expression and purification tag, pH 8.0, 50°C
Thermotoga maritima
2866
-
xylose tetraacetate
mutant lacking 10 amino acids at N-terminus, pH 8.0, 50°C
Thermotoga maritima
Other publictions for EC 3.1.1.41
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)
749261
Singh
Structural role of a conserve ...
Thermotoga maritima, Thermotoga maritima ATCC 43589, Thermotoga maritima DSM 3109
Proteins
85
694-708
2017
-
-
1
1
2
-
-
18
-
-
2
-
-
6
-
-
1
-
-
-
13
-
23
-
3
-
1
20
1
1
-
-
-
-
-
-
-
1
-
1
2
-
-
-
-
18
-
-
2
-
-
-
-
1
-
-
13
-
23
-
3
-
1
20
1
1
-
-
-
-
-
-
19
19
746975
Singh
Crystal structure of Thermoto ...
Thermotoga maritima, Thermotoga maritima ATCC 43589, Thermotoga maritima DSM 3109
Biochem. Biophys. Res. Commun.
476
63-68
2016
-
-
-
1
-
-
-
-
-
-
-
-
-
8
-
-
1
-
-
-
-
-
16
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
16
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
749834
Ma
-
High-level expression of Ceph ...
Bacillus subtilis, Bacillus subtilis SIL3
Biochem. Eng. J.
114
183-190
2016
-
1
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
749837
Ma
-
One-pot enzymatic production ...
Pseudomonas sp. SE83
Biochem. Eng. J.
95
1-8
2015
-
1
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
752071
Tao
-
Efficient production of perac ...
Bacillus subtilis
Process Biochem.
50
2121-2127
2015
-
-
1
-
1
-
1
5
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
1
-
5
-
-
-
-
-
-
-
-
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
5
5
729073
Tian
A novel cephalosporin deacetyl ...
Bacillus subtilis, Bacillus subtilis CICC 20034
Appl. Microbiol. Biotechnol.
98
2081-2089
2014
-
-
1
-
-
-
15
-
-
-
2
2
-
2
-
-
1
-
-
-
6
-
13
1
1
1
3
-
1
1
1
-
-
-
-
-
-
1
-
-
-
-
-
15
-
-
-
-
2
2
-
-
-
1
-
-
6
-
13
1
1
1
3
-
1
1
1
-
-
1
1
-
-
-
730072
Wang
Double knockout of beta-lactam ...
Escherichia coli, Escherichia coli JM105
J. Biosci. Bioeng.
113
737-741
2012
-
-
-
-
1
-
-
-
-
-
-
2
-
7
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
730907
Levisson
Functional and structural char ...
Thermotoga maritima
Proteins
80
1545-1559
2012
-
-
1
1
-
-
2
-
1
2
-
2
-
1
-
-
1
1
-
-
-
-
4
1
1
-
-
-
1
-
-
-
1
-
-
-
-
1
-
1
-
-
-
2
1
-
1
2
-
2
-
-
-
1
-
-
-
-
4
1
1
-
-
-
1
-
-
-
-
2
2
-
-
-
747164
Hedge
The structural basis for the ...
Thermotoga maritima, Thermotoga maritima DSM 3109
Biochim. Biophys. Acta
1824
1024-1030
2012
-
-
1
-
1
-
-
10
-
-
-
-
-
4
-
-
1
-
-
-
-
-
15
-
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
10
-
-
-
-
-
-
-
1
-
-
-
-
15
-
1
-
-
-
1
-
-
-
-
-
-
-
10
10
677538
Martinez-Martinez
A colorimetric assay for the d ...
Bacillus pumilus, Bacillus pumilus CECT 5072
Anal. Biochem.
369
210-217
2007
-
1
1
-
-
-
-
2
-
-
-
-
-
4
-
-
1
-
-
-
-
-
4
-
1
-
-
-
1
-
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
1
-
-
-
-
4
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
671502
Takimoto
Batch production of deacetyl 7 ...
Bacillus subtilis, Bacillus subtilis SHS0133
Appl. Microbiol. Biotechnol.
65
263-267
2004
-
1
1
-
-
1
1
-
-
-
-
2
-
5
-
-
1
-
-
-
-
1
4
-
-
-
1
-
-
-
2
-
-
-
-
-
1
1
-
-
-
1
-
1
-
-
-
-
-
2
-
-
-
1
-
-
-
1
4
-
-
-
1
-
-
-
2
-
-
-
-
-
-
-
675216
Basch
Expression of a cephalosporin ...
Rhodotorula toruloides
J. Ind. Microbiol. Biotechnol.
31
531-539
2004
-
1
1
-
-
-
-
-
-
-
-
1
-
4
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
652917
Vincent
Multifunctional xylooligosacch ...
Bacillus subtilis
J. Mol. Biol.
330
593-606
2003
-
-
-
1
-
-
-
-
-
-
-
-
-
3
-
-
-
1
-
-
-
-
5
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
5
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
649401
Velasco
Cloning and characterization o ...
Acremonium chrysogenum, Acremonium chrysogenum C10 / ATCC 48272
Appl. Microbiol. Biotechnol.
57
350-356
2001
-
-
-
-
-
-
2
1
1
-
2
-
-
7
-
1
1
-
-
-
-
-
4
-
1
1
-
-
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
2
-
1
1
-
2
-
-
-
1
1
-
-
-
-
4
-
1
1
-
-
1
1
-
1
-
-
-
-
-
-
651823
Lee
-
Isolation, analysis, and expre ...
Staphylococcus sp.
J. Biochem. Mol. Biol.
34
274-277
2001
-
-
1
-
-
-
-
-
-
-
1
-
-
1
-
1
1
-
-
-
-
-
2
-
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
1
1
-
-
-
-
2
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
649397
Cardoza
A cephalosporin C acetylhydrol ...
Amycolatopsis lactamdurans, Amycolatopsis lactamdurans MA4213
Appl. Microbiol. Biotechnol.
54
406-412
2000
-
-
-
-
-
-
5
2
1
-
1
-
-
4
-
-
1
-
-
1
1
-
8
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
5
-
2
1
-
1
-
-
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1
1
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8
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1
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170677
Takimoto
High-level expression, purific ...
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456-462
1999
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1
11
1
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1
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1
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5
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1
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1
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1
1
11
1
1
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1
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170672
Sakai
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Purification and properties of ...
Rhodotorula glutinis, Rhodotorula glutinis 38B1
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1998
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8
1
1
1
1
4
1
1
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6
-
4
1
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2
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1
1
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1
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8
1
1
1
1
4
1
1
-
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170675
Politino
Purification and characterizat ...
Bacillus subtilis, Rhodotorula toruloides
Appl. Environ. Microbiol.
63
4807-4811
1997
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3
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1
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1
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7
1
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2
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1
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1
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3
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1
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1
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1
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7
1
1
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2
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1
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170676
Sakai
Bioconversion of 7-aminocephal ...
Rhodotorula glutinis
Appl. Environ. Microbiol.
62
2669-2672
1996
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1
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1
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3
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170674
Mitsushima
Gene cloning, nucleotide seque ...
Bacillus subtilis, Bacillus subtilis SHS 0133
Appl. Environ. Microbiol.
61
2224-2229
1995
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1
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8
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170673
Takimoto
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Purification, characterization ...
Bacillus subtilis, Bacillus subtilis ATCC 6633, Bacillus subtilis SHS 0133
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1994
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7
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1
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16
2
1
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4
2
2
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1
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7
-
7
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3
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1
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1
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16
2
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4
2
2
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1
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660484
Verweij
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Industrial transformation of p ...
Escherichia coli, Pseudomonas sp., Pseudomonas sp. GK-16
Rec. Trav. Chim. Pays-Bas
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1993
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3
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2
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3
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2
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170663
Singh
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Fusarium oxysporum, Fusarium oxysporum AY F-298
Eur. J. Appl. Microbiol. Biotechnol.
9
15-18
1980
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6
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1
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170667
Abbott
Cephalosporin acetylesterase ( ...
Bacillus subtilis, Bacillus subtilis WRRL-B-558
Methods Enzymol.
43
731-734
1978
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2
3
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12
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1
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1
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1
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2
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3
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1
12
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1
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1
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1
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170664
Abbott
Immobilization of a cephalospo ...
Bacillus subtilis
Biotechnol. Bioeng.
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1033-1042
1976
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1
2
1
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1
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170665
Hinnem
Enzymatic hydrolysis of cephal ...
Acremonium chrysogenum
Antimicrob. Agents Chemother.
9
824-830
1976
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170670
Nuesch
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A biochemical and genetical ap ...
Acremonium chrysogenum
Proc. Int. Symp. Genet. Ind. Microorg. (McDonald, K. D. , ed. )
451-472
1976
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Abbott
Physical properties and kineti ...
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1975
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Abraham
Cephalosporin acetylesterase ( ...
Citrus sp.
Methods Enzymol.
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1975
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Fujisawa
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Deacetylcephalosporin C format ...
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1975
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Fujisawa
New findings on cephalosporin ...
Acremonium chrysogenum
Nature New Biol.
246
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1973
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