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Literature summary for 3.1.8.1 extracted from

  • Park, Y.J.; Yoon, S.J.; Lee, H.B.
    A novel thermostable arylesterase from the archaeon Sulfolobus solfataricus P1: purification, characterization, and expression (2008), J. Bacteriol., 190, 8086-8095.
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate the enzyme is activated by 87% by incubation with 1% (w/v) 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate, at 30°C Saccharolobus solfataricus
dimethyl sulfoxide 21% increase of activity at 50% (v/v) dimethyl sulfoxide, at 30°C Saccharolobus solfataricus
ethanol 15% increase of activity at 50% (v/v) ethanol, at 30°C Saccharolobus solfataricus
Lubrol the enzyme is activated by 14% by incubation with 1% Lubrol, at 30°C Saccharolobus solfataricus
paraoxon 115 increase of activita in the presence of 0.5 mM paraoxon, after 30 min at 30°C Saccharolobus solfataricus
pyridoxal 5'-phosphate 110% activity in the presence of 5 mM pyridoxal 5'-phosphate, after 30 min at 30°C Saccharolobus solfataricus
Tween 20 the enzyme is activated by 67% by incubation for 60 min with 1% (v/v) Tween 20 at 30°C Saccharolobus solfataricus

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Saccharolobus solfataricus

Protein Variants

Protein Variants Comment Organism
C105S mutant shows 24.0% activity compared to the wild type enzyme Saccharolobus solfataricus
C107S mutant shows 30.8% activity compared to the wild type enzyme Saccharolobus solfataricus
C129S mutant shows 3.6% activity compared to the wild type enzyme Saccharolobus solfataricus
D251N mutant shows 0.6% activity compared to the wild type enzyme Saccharolobus solfataricus
H281N mutant shows 19.4% activity compared to the wild type enzyme Saccharolobus solfataricus
S156A mutant shows 0.012% activity compared to the wild type enzyme Saccharolobus solfataricus

Inhibitors

Inhibitors Comment Organism Structure
2-propanol 55% residual activity in the presence of 90% (v/v) 2-propanol, after 60 min at 70°C Saccharolobus solfataricus
acetonitrile 43% residual activity in the presence of 90% (v/v) acetonitrile, after 60 min at 70°C Saccharolobus solfataricus
diethyl dicarbonate 6% residual activity in the presence of 5 mM diethyl dicarbonate, after 30 min at 30°C Saccharolobus solfataricus
diisopropylfluorophosphate complete inactivation at 5 mM diisopropylfluorophosphate, after 30 min at 30°C Saccharolobus solfataricus
dimethyl sulfoxide 71% residual activity in the presence of 90% (v/v) dimethyl sulfoxide, after 60 min at 70°C Saccharolobus solfataricus
EDTA 92% residual activity in the presence of 10 mM EDTA, after 30 min at 75°C Saccharolobus solfataricus
eserine 92% residual activity in the presence of 5 mM eserine, after 30 min at 30°C Saccharolobus solfataricus
ethanol 63% residual activity in the presence of 90% (v/v) ethanol, after 60 min at 70°C Saccharolobus solfataricus
Hg2+ complete inactivation at 5 mM Hg2+, after 30 min at 30°C Saccharolobus solfataricus
methanol 48% residual activity in the presence of 90% (v/v) methanol, after 60 min at 70°C Saccharolobus solfataricus
NaCl 76% residual activity in the presence of 2 M NaCl, after 60 min at 70°C Saccharolobus solfataricus
p-chloromercuribenzoate 23% residual activity in the presence of 5 mM p-chloromercuribenzoate, after 30 min at 30°C Saccharolobus solfataricus
Phenylglyoxal 97% residual activity in the presence of 5 mM phenylglyoxal, after 30 min at 30°C Saccharolobus solfataricus
phenylmethylsulfonyl fluoride 23% residual activity in the presence of 5 mM phenylmethylsulfonyl fluoride, after 30 min at 30°C Saccharolobus solfataricus
SDS about half of the activity is retained at 1% SDS after incubation for 60 min at 70°C, the addition of 5% SDS completely eliminates the enzyme activity at 70°C Saccharolobus solfataricus
Triton X-100 11% residual activity in the presence of 5% (v/v) Tween 80, after 60 min at 70°C Saccharolobus solfataricus
Tween 80 21% residual activity in the presence of 5% (v/v) Tween 80, after 60 min at 70°C Saccharolobus solfataricus
Urea 88% residual activity in the presence of 8 M urea, after 60 min at 70°C Saccharolobus solfataricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.005
-
paraoxon recombinant wild type enzyme, in 100 mM sodium phosphate buffer (pH 7.0), at 60°C Saccharolobus solfataricus
0.246
-
methyl paraoxon recombinant wild type enzyme, in 100 mM sodium phosphate buffer (pH 7.0), at 60°C Saccharolobus solfataricus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35175
-
1 * 35175, MALDI-TOF mass spectrometry Saccharolobus solfataricus
35180
-
MALDI-TOF mass spectrometry Saccharolobus solfataricus
37000
-
SDS-PAGE Saccharolobus solfataricus

Organic Solvent Stability

Organic Solvent Comment Organism
Triton X-100 after incubation for 60 min with Triton X-100 the enzyme activity is remarkably decreased with increasing temperature and concentration Saccharolobus solfataricus
urea the purified enzyme shows high stability against urea Saccharolobus solfataricus

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus B5BLW5
-
-
Saccharolobus solfataricus P1 B5BLW5
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme is purified by DEAE-cellulose column chromatography, butyl-Sepharose column chromatography, Q-Sepharose column chromatography, and hydroxyapatite column chromatography Saccharolobus solfataricus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
methyl paraoxon + H2O
-
Saccharolobus solfataricus 4-nitrophenol + dimethyl phosphate
-
?
paraoxon + H2O paraoxon is the best substrate for the purified arylesterase Saccharolobus solfataricus diethylphosphate + 4-nitrophenol
-
?

Subunits

Subunits Comment Organism
monomer 1 * 35175, MALDI-TOF mass spectrometry Saccharolobus solfataricus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
94
-
-
Saccharolobus solfataricus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50 94 the enzyme retains 52% of its activity after 50 h of incubation at 90°C, the enzyme is rapidly inactivated above 94°C, most of the enzyme activity is maintained for 5 days at 50°C, the enzyme activity gradually decreases with time at higher temperatures, approximately 70% of the enzyme activity remains after 5 days at 70°C, and 52% of it is still preserved after 50 h at 90°C, however, at 90°C after 5 days, the enzyme is completely inactivated Saccharolobus solfataricus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
342
-
methyl paraoxon recombinant wild type enzyme, in 100 mM sodium phosphate buffer (pH 7.0), at 60°C Saccharolobus solfataricus
597
-
paraoxon recombinant wild type enzyme, in 100 mM sodium phosphate buffer (pH 7.0), at 60°C Saccharolobus solfataricus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
Saccharolobus solfataricus