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Literature summary for 3.5.1.4 extracted from

  • Fu, L.; Li, X.; Xiao, X.; Xu, J.
    Purification and characterization of a thermostable aliphatic amidase from the hyperthermophilic archaeon Pyrococcus yayanosii CH1 (2014), Extremophiles, 18, 429-440.
    View publication on PubMed
Search for more literature for 3.5.1.4

Cloned(Commentary)

Cloned (Comment) Organism
overexpression in Escherichia coli C41 (DE3) Pyrococcus yayanosii

Inhibitors

Inhibitors Comment Organism Structure
acetone 25% (v/v), 69% inhibition Pyrococcus yayanosii
Ca2+ 10 mM, 12% inhibition Pyrococcus yayanosii
Co2+ 10 mM, 41% inhibition Pyrococcus yayanosii
Cu2+ 10 mM, 69% inhibition Pyrococcus yayanosii
ethanol 25% (v/v), 45% inhibition Pyrococcus yayanosii
Fe3+ 10 mM, 68% inhibition Pyrococcus yayanosii
Isopropanol 25% (v/v), 54% inhibition Pyrococcus yayanosii
methanol 25% (v/v), 33% inhibition Pyrococcus yayanosii
Ni2+ 10 mM, 31% inhibition Pyrococcus yayanosii
Zn2+ 10 mM, 62% inhibition Pyrococcus yayanosii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
8.33
-
 acetamide pH 7.6, 60°C Pyrococcus yayanosii
10.86
-
 Acrylamide pH 7.6, 60°C Pyrococcus yayanosii
11.08
-
 hexanamide pH 7.6, 60°C Pyrococcus yayanosii
15.92
-
 L-glutamine pH 7.6, 60°C Pyrococcus yayanosii

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ 10 mM, 1.15fold stimulation Pyrococcus yayanosii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
34000
-
 2 * 34000, SDS-PAGE Pyrococcus yayanosii
70000
-
 gel filtration Pyrococcus yayanosii

Organism

Organism UniProt Comment Textmining
Pyrococcus yayanosii F8AEP0
-
-
Pyrococcus yayanosii CH1 F8AEP0
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pyrococcus yayanosii

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
67
-
 acrylamide: pH 7.6, 60°C Pyrococcus yayanosii
143
-
 hexanamide: pH 7.6, 60°C Pyrococcus yayanosii
1000
-
 substrate: acetamide, pH 7.6, 60°C Pyrococcus yayanosii
1430
-
 L-glutamine: pH 7.6, 60°C Pyrococcus yayanosii

Storage Stability

Storage Stability Organism
4°C, the purified enzyme can be stored for longer than 6 months at in 50 mM phosphate buffer and 10% glycerol without any loss of activity Pyrococcus yayanosii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetamide + H2O
-
 Pyrococcus yayanosii acetate + NH3
-
 ?
acrylamide + H2O
-
 Pyrococcus yayanosii acrylic acid + NH3
-
 ?
acrylamide + H2O
-
 Pyrococcus yayanosii CH1 acrylic acid + NH3
-
 ?
formamide + H2O 22.5% of the activity with acetamide Pyrococcus yayanosii formate + NH3
-
 ?
formamide + H2O 22.5% of the activity with acetamide Pyrococcus yayanosii CH1 formate + NH3
-
 ?
hexanamide + H2O 38.7% of the activity with acetamide Pyrococcus yayanosii hexanoate + NH3
-
 ?
L-asparagine + H2O 4.8% of the activity with acetamide Pyrococcus yayanosii L-aspartate + NH3
-
 ?
L-asparagine + H2O 4.8% of the activity with acetamide Pyrococcus yayanosii CH1 L-aspartate + NH3
-
 ?
L-glutamine + H2O 73% of the activity with acetamide Pyrococcus yayanosii L-glutamate + NH3
-
 ?
L-glutamine + H2O 73% of the activity with acetamide Pyrococcus yayanosii CH1 L-glutamate + NH3
-
 ?
additional information no activity with: urea, benzamide, pyrazinamide, nicotinamide, cinnamamide, salicylamide Pyrococcus yayanosii ?
-
 ?
additional information no activity with: urea, benzamide, pyrazinamide, nicotinamide, cinnamamide, salicylamide Pyrococcus yayanosii CH1 ?
-
 ?

Subunits

Subunits Comment Organism
dimer 2 * 34000, SDS-PAGE Pyrococcus yayanosii

Synonyms

Synonyms Comment Organism
AmiE
-
 Pyrococcus yayanosii
PYCH_10400 locus name Pyrococcus yayanosii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
85
-
 with acetamide as the substrate Pyrococcus yayanosii

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
50 95 50°C: 53% of maximal activity, 60°C: about 80% of maximal activity, 70°C: about 90% of maximal activity, 95°C: about 89% of maximal activity Pyrococcus yayanosii

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
80
-
 t1/2: 110 min Pyrococcus yayanosii
90
-
 t1/2: 40 min Pyrococcus yayanosii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
333.3
-
 Acrylamide pH 7.6, 60°C Pyrococcus yayanosii
721.7
-
 hexanamide pH 7.6, 60°C Pyrococcus yayanosii
5051
-
 acetamide pH 7.6, 60°C Pyrococcus yayanosii
7216
-
 L-glutamine pH 7.6, 60°C Pyrococcus yayanosii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
 with acetamide as the substrate Pyrococcus yayanosii

pH Range

pH Minimum pH Maximum Comment Organism
3 8 pH 3.0-5.0: about 50% of maximal activity, pH 8.0: about 65% of maximal activity Pyrococcus yayanosii

pH Stability

pH Stability pH Stability Maximum Comment Organism
6
-
 2 h, stable Pyrococcus yayanosii
7
-
 120 min, over 60% of maximal activity is retained Pyrococcus yayanosii
8
-
 120 min, over 60% of maximal activity is retained Pyrococcus yayanosii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
30.69
-
 Acrylamide pH 7.6, 60°C Pyrococcus yayanosii
65.14
-
 hexanamide pH 7.6, 60°C Pyrococcus yayanosii
453.2
-
 L-glutamine pH 7.6, 60°C Pyrococcus yayanosii
606.3
-
 acetamide pH 7.6, 60°C Pyrococcus yayanosii