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Literature summary 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

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.1.1.1 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
3.1.1.1 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)

EC Number Cloned (Comment) Organism
3.1.1.1 expression in Escherichia coli Archaeoglobus fulgidus

Protein Variants

EC Number Protein Variants Comment Organism
3.1.1.1 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
3.1.1.1 V138G/L200R mutant enzyme shows improved enantioselectivity. Thermostability and resistance to additives or organic solvents is similar to wild-type enzyme Archaeoglobus fulgidus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.1.1.1 1-butanol 10% (v/v), complete loss of activity, wild-type enzyme and mutant enzymes V138G and V138G/L200R Archaeoglobus fulgidus
3.1.1.1 1-propanol 10% (v/v), complete loss of activity, wild-type enzyme and mutant enzymes V138G and V138G/L200R Archaeoglobus fulgidus
3.1.1.1 acetone 20% (v/v), complete loss of activity, wild-type enzyme and mutant enzymes V138G and V138G/L200R Archaeoglobus fulgidus
3.1.1.1 acetonitrile 20% (v/v), complete loss of activity, wild-type enzyme and mutant enzymes V138G and V138G/L200R Archaeoglobus fulgidus
3.1.1.1 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
3.1.1.1 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
3.1.1.1 ethanol 20% (v/v), complete loss of activity, wild-type enzyme and mutant enzymes V138G and V138G/L200R Archaeoglobus fulgidus
3.1.1.1 Isopropanol 20% (v/v), complete loss of activity, wild-type enzyme and mutant enzymes V138G and V138G/L200R Archaeoglobus fulgidus
3.1.1.1 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
3.1.1.1 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.1.1 3.1
-
(S)-ketoprofen ethyl ester pH 8.0, 80°C, wild-type enzyme Archaeoglobus fulgidus
3.1.1.1 3.9
-
(S)-ketoprofen ethyl ester pH 6.0, 80°C, mutant enzyme V138G/L200R Archaeoglobus fulgidus
3.1.1.1 20.9
-
(S)-ketoprofen ethyl ester pH 8.0, 80°C, mutant enzyme V138G Archaeoglobus fulgidus

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.1 Archaeoglobus fulgidus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.1.1
-
Archaeoglobus fulgidus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.1 (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
-
?
3.1.1.1 (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

EC Number Synonyms Comment Organism
3.1.1.1 Est-AF
-
Archaeoglobus fulgidus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.1.1.1 70
-
assay at Archaeoglobus fulgidus
3.1.1.1 80
-
-
Archaeoglobus fulgidus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.1.1.1 50 95 50°C: about 65% of maximal activity, 95°C: about 70% of maximal acttivity, wild-type enzyme Archaeoglobus fulgidus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.1.1.1 80
-
the enzyme activity of V138G/L200R is rapidly decreased above 80°C Archaeoglobus fulgidus
3.1.1.1 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]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1.1.1 65.4
-
(S)-ketoprofen ethyl ester pH 8.0, 80°C, wild-type enzyme Archaeoglobus fulgidus
3.1.1.1 83.9
-
(S)-ketoprofen ethyl ester pH 6.0, 80°C, mutant enzyme V138G/L200R Archaeoglobus fulgidus
3.1.1.1 385.5
-
(S)-ketoprofen ethyl ester pH 8.0, 80°C, mutant enzyme V138G Archaeoglobus fulgidus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.1.1 6
-
mutant enzyme V138G/L200R shows the highest enzyme activity at pH 6.0 Archaeoglobus fulgidus
3.1.1.1 8
-
wild-type enzyme and mutant enzyme V138G show the highest enzyme activity at pH 8.0 Archaeoglobus fulgidus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.1.1.1 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
3.1.1.1 4.8 10 pH 4.8: about 55% of maximal activity, pH 10.0: about 75% of maximal activity, wild-type enzyme Archaeoglobus fulgidus
3.1.1.1 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.1.1.1 18.5
-
(S)-ketoprofen ethyl ester pH 8.0, 80°C, mutant enzyme V138G Archaeoglobus fulgidus
3.1.1.1 21
-
(S)-ketoprofen ethyl ester pH 8.0, 80°C, wild-type enzyme Archaeoglobus fulgidus
3.1.1.1 21.3
-
(S)-ketoprofen ethyl ester pH 6.0, 80°C, mutant enzyme V138G/L200R Archaeoglobus fulgidus