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Literature summary for 3.1.1.41 extracted from

  • Singh, M.; Manoj, N.
    Structural role of a conserved active site cis proline in the Thermotoga maritima acetyl esterase from the carbohydrate esterase family 7 (2017), Proteins, 85, 694-708 .
    View publication on PubMed
Search for more literature for 3.1.1.41

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli DH5alpha cells Thermotoga maritima

Crystallization (Commentary)

Crystallization (Comment) 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 Thermotoga maritima
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

Protein Variants

Protein Variants Comment 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 Thermotoga maritima
P228A 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 Comment 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 Thermotoga maritima
0.134
-
 xylose tetraacetate 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 Thermotoga maritima
0.207
-
 Glucose pentaacetate 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] Comment Organism
76000 78000 sedimentation equilibrium centrifugation, minor peak Thermotoga maritima
210000 220000 sedimentation equilibrium centrifugation, dominant peak Thermotoga maritima

Organism

Organism UniProt Comment Textmining
Thermotoga maritima Q9WXT2
-
-
Thermotoga maritima ATCC 43589 Q9WXT2
-
-
Thermotoga maritima DSM 3109 Q9WXT2
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Thermotoga maritima

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.075
-
 substrate: 7-aminocephalosporanic acid, pH 8.0, 50°C, mutant enzyme P228A Thermotoga maritima
0.075
-
 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 Thermotoga maritima
0.47
-
 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 Thermotoga maritima
1.78
-
 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 Thermotoga maritima
3.98
-
 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 Thermotoga maritima
10
-
 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 Thermotoga maritima
25.31
-
 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 Thermotoga maritima
29.8
-
 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 Thermotoga maritima
41.49
-
 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 Thermotoga maritima
49.33
-
 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 Thermotoga maritima
346.22
-
 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 Thermotoga maritima
415.5
-
 with D-glucose pentaacetate as substrate, wild type enzyme, at pH 8.0 and 50°C Thermotoga maritima

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1-naphthyl acetate + H2O
-
 Thermotoga maritima 1-naphthol + acetate
-
 ?
1-naphthyl acetate + H2O
-
 Thermotoga maritima DSM 3109 1-naphthol + acetate
-
 ?
1-naphthyl acetate + H2O
-
 Thermotoga maritima ATCC 43589 1-naphthol + acetate
-
 ?
4-methylumbelliferyl acetate + H2O
-
 Thermotoga maritima 4-methylumbelliferol + acetate
-
 ?
4-methylumbelliferyl acetate + H2O
-
 Thermotoga maritima DSM 3109 4-methylumbelliferol + acetate
-
 ?
4-methylumbelliferyl acetate + H2O
-
 Thermotoga maritima ATCC 43589 4-methylumbelliferol + acetate
-
 ?
4-nitrophenyl acetate + H2O
-
 Thermotoga maritima 4-nitrophenol + acetate
-
 ?
4-nitrophenyl acetate + H2O
-
 Thermotoga maritima DSM 3109 4-nitrophenol + acetate
-
 ?
4-nitrophenyl acetate + H2O
-
 Thermotoga maritima ATCC 43589 4-nitrophenol + acetate
-
 ?
7-amino-cephalosporanic acid + H2O
-
 Thermotoga maritima 7-amino-3-deacetylcephalosporanic acid + acetate
-
 ?
7-aminocephalosporanic acid + H2O
-
 Thermotoga maritima acetate + deacetyl-7-aminocephalosporanic acid
-
 ?
7-aminocephalosporanic acid + H2O
-
 Thermotoga maritima DSM 3109 acetate + deacetyl-7-aminocephalosporanic acid
-
 ?
7-aminocephalosporanic acid + H2O
-
 Thermotoga maritima ATCC 43589 acetate + deacetyl-7-aminocephalosporanic acid
-
 ?
7-aminocephalosporanic acid + H2O
-
 Thermotoga maritima 7-amino-3-deacetylcephalosporanic acid + acetate
-
 ?
D-glucose pentaacetate + H2O
-
 Thermotoga maritima D-glucose + acetate
-
 ?
D-glucose pentaacetate + H2O
-
 Thermotoga maritima DSM 3109 D-glucose + acetate
-
 ?
D-glucose pentaacetate + H2O
-
 Thermotoga maritima ATCC 43589 D-glucose + acetate
-
 ?
D-glucose pentaacetate + H2O
-
 Thermotoga maritima ?
-
 ?
D-xylose tetraacetate + H2O
-
 Thermotoga maritima ?
-
 ?
D-xylose tetraacetate + H2O
-
 Thermotoga maritima D-xylose + acetate
-
 ?
fluorescein di(acetoxymethyl ether) + H2O
-
 Thermotoga maritima ?
-
 ?
glucose pentaacetate + H2O
-
 Thermotoga maritima ? + acetate
-
 ?
xylose tetraacetate + H2O
-
 Thermotoga maritima ? + acetate
-
 ?

Synonyms

Synonyms Comment Organism
ACE
-
 Thermotoga maritima
TM0077
-
 Thermotoga maritima

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment 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] Comment 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 Comment 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 Comment Organism
8
-
-
 Thermotoga maritima
8
-
 assay at Thermotoga maritima

pH Range

pH Minimum pH Maximum Comment 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 Comment 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 Thermotoga maritima
18
-
 7-aminocephalosporanic acid 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