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Literature summary for 3.4.22.8 extracted from

  • Tanaka, H.; Nariya, H.; Suzuki, M.; Houchi, H.; Tamai, E.; Miyata, S.; Okabe, A.
    High-level production and purification of clostripain expressed in a virulence-attenuated strain of Clostridium perfringens (2011), Protein Expr. Purif., 76, 83-89.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Clostridium perfringens strains 13 and 13DELTA6 Hathewaya histolytica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.3
-
Nalpha-benzoyl-L-arginine-4-nitroanilide purified recombinant enzyme, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25°C Hathewaya histolytica
0.34
-
Nalpha-benzoyl-L-arginine-4-nitroanilide commercial enzyme preparation, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25°C Hathewaya histolytica

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
14929
-
1 * 38061 + 1 * 14929, purified recombinant enzyme, MALDI-TOF mass spectrometry Hathewaya histolytica
14947
-
1 * 38067 + 1 * 14947, commercial enzyme preparation, MALDI-TOF mass spectrometry Hathewaya histolytica
14957
-
1 * 38091 + 1 * 14957, purified recombinant enzyme, calculated from amino acid sequence Hathewaya histolytica
15000
-
1 * 38000 + 1 * 15000, SDS-PAGE Hathewaya histolytica
38000
-
1 * 38000 + 1 * 15000, SDS-PAGE Hathewaya histolytica
38061
-
1 * 38061 + 1 * 14929, purified recombinant enzyme, MALDI-TOF mass spectrometry Hathewaya histolytica
38067
-
1 * 38067 + 1 * 14947, commercial enzyme preparation, MALDI-TOF mass spectrometry Hathewaya histolytica
38091
-
1 * 38091 + 1 * 14957, purified recombinant enzyme, calculated from amino acid sequence Hathewaya histolytica

Organism

Organism UniProt Comment Textmining
Hathewaya histolytica P09870
-
-
Hathewaya histolytica ATCC 19401 P09870
-
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate precipitation, hydroxyapatite column chromatography, and benzamidine-Sepharose column chromatography Hathewaya histolytica

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.022
-
commercial enzyme preparation, using acyl-L-lysine-4-nitroanilide as substrate, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25°C Hathewaya histolytica
0.039
-
purified recombinant enzyme, using acyl-L-lysine-4-nitroanilide as substrate, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25°C Hathewaya histolytica
0.119
-
culture supernatant, using Nalpha-benzoyl-L-arginine-4-nitroanilide as substrate, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25°C Hathewaya histolytica
61.2
-
recombinant enzyme after 515fold purification, using Nalpha-benzoyl-L-arginine-4-nitroanilide as substrate, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25°C Hathewaya histolytica

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acyl-L-lysine-4-nitroanilide + H2O
-
Hathewaya histolytica acyl-L-lysine + 4-nitroaniline
-
?
acyl-L-lysine-4-nitroanilide + H2O
-
Hathewaya histolytica ATCC 19401 acyl-L-lysine + 4-nitroaniline
-
?
Nalpha-benzoyl-L-arginine-4-nitroanilide + H2O the enzyme exhibits strict specificity for arginine at the P1 position, the enzyme displays 2fold higher specific activity with N-benzoyl-L-arginine-4-nitroanilide compared to acyl-L-lysine-4-nitroanilide Hathewaya histolytica Nalpha-benzoyl-L-arginine + 4-nitroaniline
-
?
Nalpha-benzoyl-L-arginine-4-nitroanilide + H2O the enzyme exhibits strict specificity for arginine at the P1 position, the enzyme displays 2fold higher specific activity with N-benzoyl-L-arginine-4-nitroanilide compared to acyl-L-lysine-4-nitroanilide Hathewaya histolytica ATCC 19401 Nalpha-benzoyl-L-arginine + 4-nitroaniline
-
?

Subunits

Subunits Comment Organism
heterodimer 1 * 38000 + 1 * 15000, SDS-PAGE Hathewaya histolytica
heterodimer 1 * 38061 + 1 * 14929, purified recombinant enzyme, MALDI-TOF mass spectrometry Hathewaya histolytica
heterodimer 1 * 38067 + 1 * 14947, commercial enzyme preparation, MALDI-TOF mass spectrometry Hathewaya histolytica
heterodimer 1 * 38091 + 1 * 14957, purified recombinant enzyme, calculated from amino acid sequence Hathewaya histolytica

Synonyms

Synonyms Comment Organism
CLO
-
Hathewaya histolytica

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
38.8
-
Nalpha-benzoyl-L-arginine-4-nitroanilide commercial enzyme preparation, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25°C Hathewaya histolytica
69.5
-
Nalpha-benzoyl-L-arginine-4-nitroanilide purified recombinant enzyme, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25°C Hathewaya histolytica

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
116
-
Nalpha-benzoyl-L-arginine-4-nitroanilide commercial enzyme preparation, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25°C Hathewaya histolytica
234
-
Nalpha-benzoyl-L-arginine-4-nitroanilide purified recombinant enzyme, in 50 mM Tris-HCl, 5 mM dithiothreitol, and 5 mM CaCl2, at pH 7.5 and 25°C Hathewaya histolytica