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Literature summary for 3.1.1.1 extracted from

  • Ohara, K.; Unno, H.; Oshima, Y.; Hosoya, M.; Fujino, N.; Hirooka, K.; Takahashi, S.; Yamashita, S.; Kusunoki, M.; Nakayama, T.
    Structural insights into the low pH adaptation of a unique carboxylesterase from Ferroplasma: altering the pH optima of two carboxylesterases (2014), J. Biol. Chem., 289, 24499-24510.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.1.1.1

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Saccharolobus shibatae
expression in Escherichia coli Ferroplasma acidiphilum

Crystallization (Commentary)

Crystallization (Comment) Organism
protein crystals are obtained at 20°C by vapor diffusion, crystal structure is determined at 1.5 A resolution Ferroplasma acidiphilum
protein crystals of wild-type enzyme and mutant enzyme Q249P/G250E are obtained at 20°C by vapor diffusion, crystal structure is determined at 1.5 A resolution Saccharolobus shibatae

Protein Variants

Protein Variants Comment Organism
A241N/Q249P/G250E/F276S kcat and kcat/Km values for 4-nitrophenyl butyrate hydrolysis are significantly lower than the values of the wild-type enzyme. The mutant shows activations by heat treatment at either 40°C or 50°C Saccharolobus shibatae
N248A/P256Q/E257G/S283F kcat and kcat/Km values for 4-nitrophenyl butyrate hydrolysis are significantly higher than the values of the wild-type enzyme. The pH-activity profile of the mutant for the hydrolysis of 4-nitrophenyl butyrate is distinct from that of the wild-type enzyme. The optimum pH for activity of the mutant (pH 8.0) is higher than that of the wild-type enzyme (pH 5.0) Ferroplasma acidiphilum
Q249P/G250E kcat and kcat/Km values for 4-nitrophenyl butyrate hydrolysis are similar to the wild-type values and significantly higher than the values of the A241N/Q249P/G250E/F276S mutant enzyme Saccharolobus shibatae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.05
-
 4-nitrophenyl butyrate pH 6.0, 35°C, mutant enzyme A241N/Q249P/G250E/F276S Saccharolobus shibatae
0.13
-
 4-nitrophenyl hexanoate pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F Ferroplasma acidiphilum
0.13
-
 4-nitrophenyl butyrate pH 6.0, 35°C, wild-type enzyme Saccharolobus shibatae
0.13
-
 4-nitrophenyl hexanoate pH 6.0, 35°C, wild-type enzyme Ferroplasma acidiphilum
0.18
-
 4-nitrophenyl butyrate pH 6.0, 35°C, mutant enzyme Q249P/G250E Saccharolobus shibatae
0.32
-
 4-nitrophenyl butyrate pH 6.0, 35°C, wild-type enzyme Ferroplasma acidiphilum
0.37
-
 4-nitrophenyl propionate pH 6.0, 35°C, wild-type enzyme Ferroplasma acidiphilum
0.48
-
 4-nitrophenyl butyrate pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F Ferroplasma acidiphilum
0.7
-
 4-nitrophenyl propionate pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F Ferroplasma acidiphilum
0.73
-
 4-nitrophenyl acetate pH 6.0, 35°C, wild-type enzyme Ferroplasma acidiphilum
1.2
-
 4-nitrophenyl acetate pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F Ferroplasma acidiphilum

Metals/Ions

Metals/Ions Comment Organism Structure
additional information the enzyme does not bind iron atoms Ferroplasma acidiphilum

Organism

Organism UniProt Comment Textmining
Ferroplasma acidiphilum Q2PCE5
-
-
Saccharolobus shibatae Q5NU42
-
-
Saccharolobus shibatae DSM 5389 Q5NU42
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Saccharolobus shibatae
-
 Ferroplasma acidiphilum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl acetate + H2O
-
 Ferroplasma acidiphilum 4-nitrophenol + acetate
-
 ?
4-nitrophenyl butyrate + H2O
-
 Saccharolobus shibatae 4-nitrophenol + butyrate
-
 ?
4-nitrophenyl butyrate + H2O
-
 Ferroplasma acidiphilum 4-nitrophenol + butyrate
-
 ?
4-nitrophenyl butyrate + H2O
-
 Saccharolobus shibatae DSM 5389 4-nitrophenol + butyrate
-
 ?
4-nitrophenyl hexanoate + H2O
-
 Ferroplasma acidiphilum 4-nitrophenol + hexanoate
-
 ?
4-nitrophenyl propionate + H2O
-
 Ferroplasma acidiphilum 4-nitrophenol + propionate
-
 ?

Synonyms

Synonyms Comment Organism
EStFa
-
 Ferroplasma acidiphilum
SshEstI
-
 Saccharolobus shibatae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
35
-
 assay at Ferroplasma acidiphilum
50
-
 hydrolysis of 4-nitrophenol butyrate, wild-type enzyme and mutant enzyme exceeded 120 min Ferroplasma acidiphilum

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
20 55 20°C: about 40% of maximal activity, 55°C: about 55% of maximal activity, hydrolysis of 4-nitrophenol butyrate, wild-type enzyme Ferroplasma acidiphilum

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
 the enzyme is inactivated at 50°C in biphasic kinetics where the t1/2 value of the fast phase of inactivation is less than 1 min. The half-life of mutant enzyme N248A/P256Q/E257G/S283F exceeds 120 min Ferroplasma acidiphilum
60
-
 half-life of mutant enzyme A241N/Q249P/G250E/F276S is 30 min Saccharolobus shibatae
90
-
 half-life of wild-type enzyme is greater than 60 min Saccharolobus shibatae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2 8 4-nitrophenyl propionate pH 6.0, 35°C, wild-type enzyme Ferroplasma acidiphilum
6
-
 4-nitrophenyl butyrate pH 6.0, 35°C, mutant enzyme A241N/Q249P/G250E/F276S Saccharolobus shibatae
11
-
 4-nitrophenyl hexanoate pH 6.0, 35°C, wild-type enzyme Ferroplasma acidiphilum
13
-
 4-nitrophenyl acetate pH 6.0, 35°C, wild-type enzyme Ferroplasma acidiphilum
31
-
 4-nitrophenyl acetate pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F Ferroplasma acidiphilum
56
-
 4-nitrophenyl butyrate pH 6.0, 35°C, wild-type enzyme Ferroplasma acidiphilum
59
-
 4-nitrophenyl propionate pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F Ferroplasma acidiphilum
101
-
 4-nitrophenyl butyrate pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F Ferroplasma acidiphilum
159
-
 4-nitrophenyl hexanoate pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F Ferroplasma acidiphilum
859
-
 4-nitrophenyl butyrate pH 6.0, 35°C, wild-type enzyme Saccharolobus shibatae
872
-
 4-nitrophenyl butyrate pH 6.0, 35°C, mutant enzyme Q249P/G250E Saccharolobus shibatae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5
-
 hydrolysis of 4-nitrophenol butyrate, wild-type enzyme Ferroplasma acidiphilum
6
-
 assay at Ferroplasma acidiphilum
8
-
-
 Saccharolobus shibatae
8
-
 hydrolysis of 4-nitrophenol butyrate, mutant enzyme N248A/P256Q/E257G/S283F Ferroplasma acidiphilum

pH Range

pH Minimum pH Maximum Comment Organism
3 8 pH 3.0: about 60% of maximal activity, pH 8.0: about 60% of maximal activity, hydrolysis of 4-nitrophenol butyrate, wild-type enzyme Ferroplasma acidiphilum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
17.8
-
 4-nitrophenyl acetate pH 6.0, 35°C, wild-type enzyme Ferroplasma acidiphilum
25.8
-
 4-nitrophenyl acetate pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F Ferroplasma acidiphilum
74.4
-
 4-nitrophenyl propionate pH 6.0, 35°C, wild-type enzyme Ferroplasma acidiphilum
79.5
-
 4-nitrophenyl hexanoate pH 6.0, 35°C, wild-type enzyme Ferroplasma acidiphilum
84.6
-
 4-nitrophenyl propionate pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F Ferroplasma acidiphilum
119
-
 4-nitrophenyl butyrate pH 6.0, 35°C, mutant enzyme A241N/Q249P/G250E/F276S Saccharolobus shibatae
173
-
 4-nitrophenyl butyrate pH 6.0, 35°C, wild-type enzyme Ferroplasma acidiphilum
211
-
 4-nitrophenyl butyrate pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F Ferroplasma acidiphilum
1275
-
 4-nitrophenyl hexanoate pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F Ferroplasma acidiphilum
4807
-
 4-nitrophenyl butyrate pH 6.0, 35°C, mutant enzyme Q249P/G250E Saccharolobus shibatae
6850
-
 4-nitrophenyl butyrate pH 6.0, 35°C, wild-type enzyme Saccharolobus shibatae