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Literature summary for 3.4.21.26 extracted from

  • Tokai, S.; Bito, T.; Shimizu, K.; Arima, J.
    Methionine residues lining the substrate pathway in prolyl oligopeptidase from Pleurotus eryngii play an important role in substrate recognition (2018), Biosci. Biotechnol. Biochem., 82, 1107-1115 .
    View publication on PubMed
Search for more literature for 3.4.21.26

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rosetta (DE3) cells Pleurotus eryngii

Protein Variants

Protein Variants Comment Organism
M142A the mutant exhibits a wild type Phe/Leu ratio and has overall reduced activity compared to the wild type enzyme Pleurotus eryngii
M142Q/M203Q/M570Q/M620Q the mutant shows an activity ratio (23.6) and pattern of substrate specificity essentially identical to that of oxidized enzyme Pleurotus eryngii
M203A the mutant shows a Phe/Leu ratio (14.5) higher than that of wild type enzyme. Oxidation of Met203 on the non-catalytic beta-propeller domain with 1% (v/v) H2O2 results in decreased activity toward non-aromatic aminoacyl-4-nitroanilides while maintaining its activity toward aromatic aminoacyl-4-nitroanilides Pleurotus eryngii
M203Q the mutant shows 4-nitroanilide-release activity lower than that of wild type enzyme and a Phe/Leu ratio of 7.4 compared to the wild type Pleurotus eryngii
M203Q/M570Q the mutant shows 4-nitroanilide-release activity lower than that of wild type enzyme and a Phe/Leu ratio (15.4) higher than those of M203Q and M570Q mutant enzymes Pleurotus eryngii
M570A the mutant has overall reduced activity compared to the wild type enzyme and shows a Phe/Leu ratio (15.6) higher than that of wild type enzyme. The mutation is responsible for the altered substrate specificity of the enzyme following oxidation Pleurotus eryngii
M570E the mutant shows 4-nitroanilide-release activity lower than that of wild type enzyme Pleurotus eryngii
M570F the mutant shows 4-nitroanilide-release activity lower than that of wild type enzyme Pleurotus eryngii
M570K the mutant shows 4-nitroanilide-release activity lower than that of wild type enzyme Pleurotus eryngii
M570Q the mutant shows 4-nitroanilide-release activity lower than that of wild type enzyme and a Phe/Leu ratio of 9.3 compared to the wild type Pleurotus eryngii
M620T the mutant exhibits a wild type Phe/Leu ratio and has overall reduced activity compared to the wild type enzyme Pleurotus eryngii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.063
-
 L-Phe-4-nitroanilide wild type enzyme, at pH 8.0 and 25°C Pleurotus eryngii
0.11
-
 L-Phe-4-nitroanilide mutant enzyme M570E, at pH 8.0 and 25°C Pleurotus eryngii
0.12
-
 L-Leu-4-nitroanilide wild type enzyme, at pH 8.0 and 25°C Pleurotus eryngii
0.12
-
 L-Phe-4-nitroanilide mutant enzyme M142Q/M203Q/M570Q/M620Q, at pH 8.0 and 25°C Pleurotus eryngii
0.16
-
 L-Phe-4-nitroanilide mutant enzyme M570A, at pH 8.0 and 25°C Pleurotus eryngii
0.18
-
 L-Phe-4-nitroanilide mutant enzyme M203Q, at pH 8.0 and 25°C Pleurotus eryngii
0.18
-
 L-Phe-4-nitroanilide mutant enzyme M570Q, at pH 8.0 and 25°C Pleurotus eryngii
0.3
-
 L-Leu-4-nitroanilide mutant enzyme M203Q, at pH 8.0 and 25°C Pleurotus eryngii
0.34
-
 L-Phe-4-nitroanilide mutant enzyme M203Q/M570Q, at pH 8.0 and 25°C Pleurotus eryngii
0.62
-
 L-Leu-4-nitroanilide mutant enzyme M203Q/M570Q, at pH 8.0 and 25°C Pleurotus eryngii
0.64
-
 L-Phe-4-nitroanilide mutant enzyme M570F, at pH 8.0 and 25°C Pleurotus eryngii
0.68
-
 L-Leu-4-nitroanilide mutant enzyme M142Q/M203Q/M570Q/M620Q, at pH 8.0 and 25°C Pleurotus eryngii
0.72
-
 L-Leu-4-nitroanilide oxidized wild type enzyme, at pH 8.0 and 25°C Pleurotus eryngii
0.73
-
 L-Leu-4-nitroanilide mutant enzyme M570F, at pH 8.0 and 25°C Pleurotus eryngii
0.82
-
 L-Phe-4-nitroanilide mutant enzyme M570K, at pH 8.0 and 25°C Pleurotus eryngii
0.86
-
 L-Leu-4-nitroanilide mutant enzyme M570K, at pH 8.0 and 25°C Pleurotus eryngii
0.89
-
 L-Leu-4-nitroanilide mutant enzyme M570Q, at pH 8.0 and 25°C Pleurotus eryngii
0.98
-
 L-Leu-4-nitroanilide mutant enzyme M570A, at pH 8.0 and 25°C Pleurotus eryngii
1.1
-
 L-Leu-4-nitroanilide mutant enzyme M570E, at pH 8.0 and 25°C Pleurotus eryngii
6.2
-
 L-Phe-4-nitroanilide oxidized wild type enzyme, at pH 8.0 and 25°C Pleurotus eryngii

Organism

Organism UniProt Comment Textmining
Pleurotus eryngii
-
-
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate precipitation and Butyl Toyopearl column chromatography Pleurotus eryngii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-Ala-4-nitroanilide + H2O
-
 Pleurotus eryngii L-Ala + 4-nitroaniline
-
 ?
L-His-4-nitroanilide + H2O worst substrate Pleurotus eryngii L-His + 4-nitroaniline
-
 ?
L-Leu-4-nitroanilide + H2O
-
 Pleurotus eryngii L-Leu + 4-nitroaniline
-
 ?
L-Met-4-nitroanilide + H2O
-
 Pleurotus eryngii L-Met + 4-nitroaniline
-
 ?
L-Phe-4-nitroanilide + H2O best substrate Pleurotus eryngii L-Phe + 4-nitroaniline
-
 ?
L-Pro-4-nitroanilide + H2O
-
 Pleurotus eryngii L-Pro + 4-nitroaniline
-
 ?
L-Tyr-4-nitroanilide + H2O second best substrate Pleurotus eryngii L-Tyr + 4-nitroaniline
-
 ?

Synonyms

Synonyms Comment Organism
eryngase
-
 Pleurotus eryngii
POP
-
 Pleurotus eryngii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.9
-
 L-Leu-4-nitroanilide mutant enzyme M570K, at pH 8.0 and 25°C Pleurotus eryngii
6.4
-
 L-Leu-4-nitroanilide oxidized wild type enzyme, at pH 8.0 and 25°C Pleurotus eryngii
26
-
 L-Leu-4-nitroanilide mutant enzyme M570F, at pH 8.0 and 25°C Pleurotus eryngii
58
-
 L-Leu-4-nitroanilide mutant enzyme M203Q/M570Q, at pH 8.0 and 25°C Pleurotus eryngii
75
-
 L-Leu-4-nitroanilide mutant enzyme M142Q/M203Q/M570Q/M620Q, at pH 8.0 and 25°C Pleurotus eryngii
180
-
 L-Leu-4-nitroanilide mutant enzyme M570Q, at pH 8.0 and 25°C Pleurotus eryngii
210
-
 L-Leu-4-nitroanilide mutant enzyme M203Q, at pH 8.0 and 25°C Pleurotus eryngii
210
-
 L-Leu-4-nitroanilide mutant enzyme M570A, at pH 8.0 and 25°C Pleurotus eryngii
240
-
 L-Leu-4-nitroanilide mutant enzyme M570E, at pH 8.0 and 25°C Pleurotus eryngii
550
-
 L-Phe-4-nitroanilide mutant enzyme M142Q/M203Q/M570Q/M620Q, at pH 8.0 and 25°C Pleurotus eryngii
570
-
 L-Leu-4-nitroanilide wild type enzyme, at pH 8.0 and 25°C Pleurotus eryngii
710
-
 L-Phe-4-nitroanilide mutant enzyme M570Q, at pH 8.0 and 25°C Pleurotus eryngii
920
-
 L-Phe-4-nitroanilide mutant enzyme M570K, at pH 8.0 and 25°C Pleurotus eryngii
950
-
 L-Phe-4-nitroanilide mutant enzyme M570E, at pH 8.0 and 25°C Pleurotus eryngii
1100
-
 L-Phe-4-nitroanilide mutant enzyme M570A, at pH 8.0 and 25°C Pleurotus eryngii
1100
-
 L-Phe-4-nitroanilide mutant enzyme M570F, at pH 8.0 and 25°C Pleurotus eryngii
1200
-
 L-Phe-4-nitroanilide mutant enzyme M203Q/M570Q, at pH 8.0 and 25°C Pleurotus eryngii
1900
-
 L-Phe-4-nitroanilide wild type enzyme, at pH 8.0 and 25°C Pleurotus eryngii
1900
-
 L-Phe-4-nitroanilide mutant enzyme M203Q, at pH 8.0 and 25°C Pleurotus eryngii
4300
-
 L-Phe-4-nitroanilide oxidized wild type enzyme, at pH 8.0 and 25°C Pleurotus eryngii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.7
-
 L-Leu-4-nitroanilide mutant enzyme M570K, at pH 8.0 and 25°C Pleurotus eryngii
8.9
-
 L-Leu-4-nitroanilide oxidized wild type enzyme, at pH 8.0 and 25°C Pleurotus eryngii
35
-
 L-Leu-4-nitroanilide mutant enzyme M570F, at pH 8.0 and 25°C Pleurotus eryngii
94
-
 L-Leu-4-nitroanilide mutant enzyme M203Q/M570Q, at pH 8.0 and 25°C Pleurotus eryngii
110
-
 L-Leu-4-nitroanilide mutant enzyme M142Q/M203Q/M570Q/M620Q, at pH 8.0 and 25°C Pleurotus eryngii
200
-
 L-Leu-4-nitroanilide mutant enzyme M570Q, at pH 8.0 and 25°C Pleurotus eryngii
220
-
 L-Leu-4-nitroanilide mutant enzyme M570A, at pH 8.0 and 25°C Pleurotus eryngii
220
-
 L-Leu-4-nitroanilide mutant enzyme M570E, at pH 8.0 and 25°C Pleurotus eryngii
700
-
 L-Phe-4-nitroanilide oxidized wild type enzyme, at pH 8.0 and 25°C Pleurotus eryngii
720
-
 L-Leu-4-nitroanilide mutant enzyme M203Q, at pH 8.0 and 25°C Pleurotus eryngii
1100
-
 L-Phe-4-nitroanilide mutant enzyme M570K, at pH 8.0 and 25°C Pleurotus eryngii
1800
-
 L-Phe-4-nitroanilide mutant enzyme M570F, at pH 8.0 and 25°C Pleurotus eryngii
3500
-
 L-Phe-4-nitroanilide mutant enzyme M203Q/M570Q, at pH 8.0 and 25°C Pleurotus eryngii
3900
-
 L-Phe-4-nitroanilide mutant enzyme M570Q, at pH 8.0 and 25°C Pleurotus eryngii
4600
-
 L-Phe-4-nitroanilide mutant enzyme M142Q/M203Q/M570Q/M620Q, at pH 8.0 and 25°C Pleurotus eryngii
4900
-
 L-Leu-4-nitroanilide wild type enzyme, at pH 8.0 and 25°C Pleurotus eryngii
6800
-
 L-Phe-4-nitroanilide mutant enzyme M570A, at pH 8.0 and 25°C Pleurotus eryngii
8300
-
 L-Phe-4-nitroanilide mutant enzyme M570E, at pH 8.0 and 25°C Pleurotus eryngii
10000
-
 L-Phe-4-nitroanilide mutant enzyme M203Q, at pH 8.0 and 25°C Pleurotus eryngii
30000
-
 L-Phe-4-nitroanilide wild type enzyme, at pH 8.0 and 25°C Pleurotus eryngii