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Literature summary for 3.4.19.1 extracted from

  • Ishikawa, K.; Ishida, H.; Koyama, Y.; Kawarabayasi, Y.; Kawahara, J.; Matsui, E.; Matsui, I.
    Acylamino acid-releasing enzyme from the thermophilic archaeon Pyrococcus horikoshii (1998), J. Biol. Chem., 273, 17726-17731 .
    View publication on PubMed
Search for more literature for 3.4.19.1

Application

Application Comment Organism
analysis the enzyme is expected to be useful for the removal of Nalpha-acylated residues in short peptide sequence analysis at high temperatures Pyrococcus horikoshii

Cloned(Commentary)

Cloned (Comment) Organism
overexpressed in Escherichia coli BL21(DE3) Pyrococcus horikoshii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
7.6
-
 Acetyl-Ala-Ala pH 5.4, 85°C Pyrococcus horikoshii
8.6
-
 Acetyl-Ala-Ala pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
11
-
 acetyl-Leu-4-nitroanilide pH 5.4, 85°C Pyrococcus horikoshii
12.9
-
 acetyl-Leu-4-nitroanilide pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
13
-
 Acetyl-Ala-Ala-Ala-Ala pH 5.4, 85°C Pyrococcus horikoshii
15.2
-
 Acetyl-Ala-Ala-Ala-Ala pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
18.4
-
 acetyl-Ala-4-nitroanilide pH 5.4, 85°C Pyrococcus horikoshii
20.2
-
 acetyl-Ala-4-nitroanilide pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
70000
-
 gel filtration Pyrococcus horikoshii

Organism

Organism UniProt Comment Textmining
Pyrococcus horikoshii AB009494
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pyrococcus horikoshii

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 the specific activity of the enzyme increases 7–14fold with heat treatment, suggesting the modification of the structure of the enzyme for optimal hydrolytic activity by heating Pyrococcus horikoshii
1.9
-
 substrate: acetyl-Tyr-4-nitroanilide, pH 5.4, 85°C Pyrococcus horikoshii
6.9
-
 substrate: acetyl-Ala-4-nitroanilide, pH 5.4, 85°C Pyrococcus horikoshii
9.5
-
 substrate: formyl-Met-Ala-Ala-Ala-Ala-Ala, pH 5.4, 85°C Pyrococcus horikoshii
10.2
-
 substrate: formyl-Ala-Ala-Ala-Ala-Ala-Ala, pH 5.4, 85°C Pyrococcus horikoshii
11.7
-
 substrate: acetyl-Met-Ala-Ala-Ala-Ala-Ala, pH 5.4, 85°C Pyrococcus horikoshii
13.6
-
 substrate: acetyl-Ala-Ala-Ala-Ala-Ala-Ala, pH 5.4, 85°C Pyrococcus horikoshii
15
-
 substrate: acetyl-Tyr-4-nitroanilide, pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
51
-
 substrate: acetyl-Ala-4-nitroanilide, pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
66.8
-
 substrate: formyl-Met-Ala-Ala-Ala-Ala-Ala, pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
77
-
 substrate: acetyl-Met-Ala-Ala-Ala-Ala-Ala, pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
77.3
-
 substrate: formyl-Ala-Ala-Ala-Ala-Ala-Ala, pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
97.2
-
 substrate: formyl-Met-Ala, pH 5.4, 85°C Pyrococcus horikoshii
105
-
 substrate: acetyl-Leu-4-nitroanilide, pH 5.4, 85°C Pyrococcus horikoshii
118
-
 substrate: acetyl-Ala-Ala-Ala-Ala-Ala-Ala, pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
206
-
 substrate: acetyl-Ala-Ala-Ala-Ala, pH 5.4, 85°C Pyrococcus horikoshii
344
-
 substrate: acetyl-Met-Ala, pH 5.4, 85°C Pyrococcus horikoshii
402
-
 substrate: formyl-Ala-Ala-Ala, pH 5.4, 85°C Pyrococcus horikoshii
478
-
 substrate: acetyl-Ala-Ala, pH 5.4, 85°C Pyrococcus horikoshii
694
-
 substrate: formyl-Met-Leu-Gly, pH 5.4, 85°C Pyrococcus horikoshii
750
-
 substrate: acetyl-Leu-4-nitroanilide, pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
1388
-
 substrate: formyl-Met-Ala-Ser, pH 5.4, 85°C Pyrococcus horikoshii
1513
-
 substrate: formyl-Met-Ala, pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
1830
-
 substrate: acetyl-Ala-Ala-Ala-Ala, pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
5120
-
 substrate: acetyl-Met-Ala, pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
5363
-
 substrate: formyl-Ala-Ala-Ala, pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
7060
-
 substrate: acetyl-Ala-Ala, pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
7064
-
 substrate: formyl-Met-Leu-Gly, pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
17026
-
 substrate: formyl-Met-Ala-Ser, pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetyl-Ala-4-nitroanilide + H2O the enzyme releases acetyl-Leu better than acetyl-Ala from acetyl-amino acid-4-nitroanilides Pyrococcus horikoshii acetyl-Ala + 4-nitroaniline
-
 ?
acetyl-Ala-Ala + H2O
-
 Pyrococcus horikoshii acetyl-Ala + Ala
-
 ?
acetyl-Ala-Ala-Ala-Ala + H2O
-
 Pyrococcus horikoshii acetyl-Ala + Ala-Ala-Ala
-
 ?
acetyl-Ala-Ala-Ala-Ala-Ala-Ala + H2O
-
 Pyrococcus horikoshii acetyl-Ala + Ala-Ala-Ala-Ala-Ala
-
 ?
acetyl-Leu-4-nitroanilide + H2O the enzyme releases acetyl-Leu better than acetyl-Ala from acetyl-amino acid-4-nitroanilides Pyrococcus horikoshii acetyl-Leu + 4-nitroaniline
-
 ?
acetyl-Met-Ala + H2O
-
 Pyrococcus horikoshii acetyl-Met + Ala
-
 ?
acetyl-Met-Ala-Ala-Ala-Ala-Ala + H2O
-
 Pyrococcus horikoshii acetyl-Met + Ala-Ala-Ala-Ala-Ala
-
 ?
acetyl-Tyr-4-nitroanilide + H2O
-
 Pyrococcus horikoshii acetyl-Tyr + 4-nitroaniline
-
 ?
formyl-Ala-Ala-Ala + H2O
-
 Pyrococcus horikoshii formyl-Ala + Ala-Ala
-
 ?
formyl-Ala-Ala-Ala-Ala-Ala-Ala + H2O
-
 Pyrococcus horikoshii formyl-Ala + Ala-Ala-Ala-Ala-Ala
-
 ?
formyl-Met-Ala + H2O
-
 Pyrococcus horikoshii formyl-Met + Ala
-
 ?
formyl-Met-Ala-Ala-Ala-Ala-Ala + H2O
-
 Pyrococcus horikoshii formyl-Met + Ala-Ala-Ala-Ala-Ala
-
 ?
formyl-Met-Ala-Ser + H2O
-
 Pyrococcus horikoshii formyl-Met + Ala-Ser
-
 ?
formyl-Met-Leu-Gly + H2O
-
 Pyrococcus horikoshii formyl-Met + Leu-Gly
-
 ?
additional information high hydrolytic activity for acylpeptides, no hydrolytic activity for Leu-4-nitroanilide and Ala-4-nitroanilide Pyrococcus horikoshii ?
-
 ?

Subunits

Subunits Comment Organism
? x * 70000, SDS-PAGE Pyrococcus horikoshii

Synonyms

Synonyms Comment Organism
AARE
-
 Pyrococcus horikoshii
acylamino acid-releasing enzyme
-
 Pyrococcus horikoshii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
additional information
-
 the specific activity of the enzyme increases 7–14fold with heat treatment, suggesting the modification of the structure of the enzyme for optimal hydrolytic activity by heating Pyrococcus horikoshii
85
-
 assay at Pyrococcus horikoshii
90
-
-
 Pyrococcus horikoshii

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
90
-
 stable for 24 h Pyrococcus horikoshii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
17.3
-
 acetyl-Ala-4-nitroanilide pH 5.4, 85°C Pyrococcus horikoshii
144
-
 acetyl-Ala-4-nitroanilide pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
281
-
 Acetyl-Ala-Ala-Ala-Ala pH 5.4, 85°C Pyrococcus horikoshii
330
-
 acetyl-Leu-4-nitroanilide pH 5.4, 85°C Pyrococcus horikoshii
790
-
 Acetyl-Ala-Ala pH 5.4, 85°C Pyrococcus horikoshii
2090
-
 Acetyl-Ala-Ala-Ala-Ala pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
3200
-
 acetyl-Leu-4-nitroanilide pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii
9900
-
 Acetyl-Ala-Ala pH 5.4, 85°C, heat-activated enzyme Pyrococcus horikoshii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
4.8 5.5 at 85°C Pyrococcus horikoshii
5.4
-
 assay at Pyrococcus horikoshii

General Information

General Information Comment Organism
metabolism the existence of the acylamino acid-releasing enzyme in archaea suggests that the mechanisms of protein degradation or initiation of protein synthesis or both in archaea may be similar to those in eukaryotes Pyrococcus horikoshii