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Literature summary for 3.1.1.1 extracted from

  • Kim, J.; Kim, S.; Yoon, S.; Hong, E.; Ryu, Y.
    Improved enantioselectivity of thermostable esterase from Archaeoglobus fulgidus toward (S)-ketoprofen ethyl ester by directed evolution and characterization of mutant esterases (2015), Appl. Microbiol. Biotechnol., 99, 6293-301.
    View publication on PubMed
Search for more literature for 3.1.1.1

Activating Compound

Activating Compound Comment Organism Structure
DMSO 10% (v/v), activity and enantioselectivity of mutant enzyme V138G toward (S)-ketoprofen ethyl ester is greatly increased. 10% and 20% (v/v) decreases activity of wild-type and mutant enzyme V138G/L200R Archaeoglobus fulgidus
n-hexane 20% (v/v), activity and enantioselectivity of mutant enzyme V138G toward (S)-ketoprofen ethyl ester is greatly increased. 10% (v/v), activity of wild-type enzyme and mutant enzyme V138G/L200R is increased Archaeoglobus fulgidus

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Archaeoglobus fulgidus

Protein Variants

Protein Variants Comment Organism
V138G mutant enzyme shows improved enantioselectivity. The mutant enzyme shows higher Vmax and kcat values than the wild-type enzyme, but the kcat/Km value of V138G decreases relative to Est-AF, due to an increase in its Km value Archaeoglobus fulgidus
V138G/L200R mutant enzyme shows improved enantioselectivity. Thermostability and resistance to additives or organic solvents is similar to wild-type enzyme Archaeoglobus fulgidus

Inhibitors

Inhibitors Comment Organism Structure
1-butanol 10% (v/v), complete loss of activity, wild-type enzyme and mutant enzymes V138G and V138G/L200R Archaeoglobus fulgidus
1-propanol 10% (v/v), complete loss of activity, wild-type enzyme and mutant enzymes V138G and V138G/L200R Archaeoglobus fulgidus
acetone 20% (v/v), complete loss of activity, wild-type enzyme and mutant enzymes V138G and V138G/L200R Archaeoglobus fulgidus
acetonitrile 20% (v/v), complete loss of activity, wild-type enzyme and mutant enzymes V138G and V138G/L200R Archaeoglobus fulgidus
DMF 10% (v/v) and 20% (v/v), strong decrease in activity of wild-type enzyme and mutant enzymes V138G and V138G/L200R Archaeoglobus fulgidus
DMSO 10% (v/v), activity and enantioselectivity of mutant enzyme V138G toward (S)-ketoprofen ethyl ester is greatly increased. 10% (v/v) and 20% (v/v) decreases activity of wild-type and mutant enzyme V138G/L200R Archaeoglobus fulgidus
ethanol 20% (v/v), complete loss of activity, wild-type enzyme and mutant enzymes V138G and V138G/L200R Archaeoglobus fulgidus
Isopropanol 20% (v/v), complete loss of activity, wild-type enzyme and mutant enzymes V138G and V138G/L200R Archaeoglobus fulgidus
methanol 10% (v/v) and 20% (v/v), strong decrease in activity of wild-type enzyme and mutant enzymes V138G and V138G/L200R Archaeoglobus fulgidus
n-hexane 20% (v/v), activity and enantioselectivity of mutant enzyme V138G toward (S)-ketoprofen ethyl ester is greatly increased. 10% (v/v), activity of wild-type enzyme and mutant enzyme V138G/L200R is increased Archaeoglobus fulgidus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1
-
 (S)-ketoprofen ethyl ester pH 8.0, 80°C, wild-type enzyme Archaeoglobus fulgidus
3.9
-
 (S)-ketoprofen ethyl ester pH 6.0, 80°C, mutant enzyme V138G/L200R Archaeoglobus fulgidus
20.9
-
 (S)-ketoprofen ethyl ester pH 8.0, 80°C, mutant enzyme V138G Archaeoglobus fulgidus

Organism

Organism UniProt Comment Textmining
Archaeoglobus fulgidus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Archaeoglobus fulgidus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R)-ketoprofen ethyl ester + H2O (R)-ketoprofen or (S)-ketoprofen is produced by esterase hydrolysis of the ester bond of (R,S)-ketoprofen ethyl ester Archaeoglobus fulgidus (R)-ketoprofen + ethanol
-
 ?
(S)-ketoprofen ethyl ester + H2O the wild-type enzyme shows low enantioselectivity toward (S)-ketoprofen ethyl ester. Mutant enzymes V138G and V138G/L200R shows improved enantioselectivity toward (S)-ketoprofen ethyl ester. The enantioselectivity of V138G and V138G/L200R is greatly improved in Tween 80 Archaeoglobus fulgidus (S)-ketoprofen + ethanol
-
 ?

Synonyms

Synonyms Comment Organism
Est-AF
-
 Archaeoglobus fulgidus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
 assay at Archaeoglobus fulgidus
80
-
-
 Archaeoglobus fulgidus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
50 95 50°C: about 65% of maximal activity, 95°C: about 70% of maximal acttivity, wild-type enzyme Archaeoglobus fulgidus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
80
-
 the enzyme activity of V138G/L200R is rapidly decreased above 80°C Archaeoglobus fulgidus
95
-
 the enzyme activity of wild-type enzyme and mutant enzyme V138G remains greater than 70% at 95°C after 30 min Archaeoglobus fulgidus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
65.4
-
 (S)-ketoprofen ethyl ester pH 8.0, 80°C, wild-type enzyme Archaeoglobus fulgidus
83.9
-
 (S)-ketoprofen ethyl ester pH 6.0, 80°C, mutant enzyme V138G/L200R Archaeoglobus fulgidus
385.5
-
 (S)-ketoprofen ethyl ester pH 8.0, 80°C, mutant enzyme V138G Archaeoglobus fulgidus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
 mutant enzyme V138G/L200R shows the highest enzyme activity at pH 6.0 Archaeoglobus fulgidus
8
-
 wild-type enzyme and mutant enzyme V138G show the highest enzyme activity at pH 8.0 Archaeoglobus fulgidus

pH Range

pH Minimum pH Maximum Comment Organism
4.8 9 pH 4.8: about 85% of maximal activity, pH 9.0: about 40% of maximal activity, mutant enzyme V138G/L200R Archaeoglobus fulgidus
4.8 10 pH 4.8: about 55% of maximal activity, pH 10.0: about 75% of maximal activity, wild-type enzyme Archaeoglobus fulgidus
4.8 11 pH 4.8: about 75% of maximal activity, pH 11.0: about 75% of maximal activity, mutant enzyme V138G Archaeoglobus fulgidus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
18.5
-
 (S)-ketoprofen ethyl ester pH 8.0, 80°C, mutant enzyme V138G Archaeoglobus fulgidus
21
-
 (S)-ketoprofen ethyl ester pH 8.0, 80°C, wild-type enzyme Archaeoglobus fulgidus
21.3
-
 (S)-ketoprofen ethyl ester pH 6.0, 80°C, mutant enzyme V138G/L200R Archaeoglobus fulgidus