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Literature summary extracted from

  • Oyama, H.; Abe, S.I.; Ushiyama, S.; Takahashi, S.; Oda, K.
    Identification of catalytic residues of pepstatin-insensitive carboxyl proteinases from prokaryotes by site-directed mutagenesis (1999), J. Biol. Chem., 274, 27815-27822.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.21.100 expression in Escherichia coli Pseudomonas sp.
3.4.21.101 expression in Escherichia coli Xanthomonas sp.

Protein Variants

EC Number Protein Variants Comment Organism
3.4.21.100 D124A slightly lower proteinase activity than that of wild-type enzyme Pseudomonas sp.
3.4.21.100 D170A mutant enzyme is not processed autocatalytically in Escherichia coli Pseudomonas sp.
3.4.21.100 D225A mutant enzyme shows almost the same proteinase activity as wild-type enzyme Pseudomonas sp.
3.4.21.100 D254A mutant enzyme shows almost the same proteinase activity as wild-type enzyme Pseudomonas sp.
3.4.21.100 D328A mutant enzyme is not processed autocatalytically in Escherichia coli Pseudomonas sp.
3.4.21.100 D84A proteinase activity is 0.2% of that of wild-type enzyme, mutant enzyme is very unstable Pseudomonas sp.
3.4.21.100 E217A mutant enzyme shows almost the same proteinase activity as wild-type enzyme Pseudomonas sp.
3.4.21.100 E222A mutant enzyme is not processed autocatalytically in Escherichia coli Pseudomonas sp.
3.4.21.101 D348A production of a 64000 Da precursor protein which is not processed to active enzymes under acidic conditions, no catalytic acticity detected Xanthomonas sp.
3.4.21.101 DELTA192 pro-region mutant enzyme Xanthomonas sp.
3.4.21.101 S169A production of a 64000 Da precursor protein which is not processed to active enzymes under acidic conditions, no catalytic acticity detected Xanthomonas sp.
3.4.21.101 S79A production of a 64000 Da precursor protein which is not processed to active enzymes under acidic conditions, no catalytic acticity detected Xanthomonas sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.21.100 0.0067
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme D124A Pseudomonas sp.
3.4.21.100 0.0074
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, wild-type enzyme Pseudomonas sp.
3.4.21.100 0.0091
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme D265A Pseudomonas sp.
3.4.21.100 0.0101
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme E217A Pseudomonas sp.
3.4.21.100 0.0296
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme D225A Pseudomonas sp.
3.4.21.100 0.149
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme D84A Pseudomonas sp.
3.4.21.101 0.0058
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme DELTA192 Xanthomonas sp.
3.4.21.101 0.0111
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme E230A Xanthomonas sp.
3.4.21.101 0.0188
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme E235A Xanthomonas sp.

Organism

EC Number Organism UniProt Comment Textmining
3.4.21.100 Pseudomonas sp. P42790
-
-
3.4.21.101 Xanthomonas sp.
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.21.100 recombinant mutant enzymes D124A, D225A, D254A, E217A, D84A, D170A, D328A and E222A Pseudomonas sp.
3.4.21.101 recombinant mutant enzymes S79A, S169A and D348A Xanthomonas sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.21.100 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu + H2O
-
 Pseudomonas sp. ?
-
 ?
3.4.21.101 casein + H2O
-
 Xanthomonas sp. ?
-
 ?
3.4.21.101 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu + H2O
-
 Xanthomonas sp. Lys-Pro-Ala-Leu-Phe + p-nitrophenylalanine-Arg-Leu
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.4.21.100 PCP
-
 Pseudomonas sp.
3.4.21.101 XCP
-
 Xanthomonas sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4.21.100 0.055
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme D84A Pseudomonas sp.
3.4.21.100 7
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, wild-type enzyme Pseudomonas sp.
3.4.21.100 39
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme D124A Pseudomonas sp.
3.4.21.100 56
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme E217A Pseudomonas sp.
3.4.21.100 59.3
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, wild-type enzyme Pseudomonas sp.
3.4.21.100 59.5
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme D265A Pseudomonas sp.
3.4.21.100 65.2
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme D225A Pseudomonas sp.
3.4.21.101 0.3
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme E235A Xanthomonas sp.
3.4.21.101 0.9
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme DELTA192 Xanthomonas sp.
3.4.21.101 21.8
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme E230A Xanthomonas sp.
3.4.21.101 34.9
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme E235A Xanthomonas sp.
3.4.21.101 68.4
-
 Lys-Pro-Ala-Leu-Phe-p-nitrophenylalanine-Arg-Leu pH 3.5, 37°C, mutant enzyme DELTA192 Xanthomonas sp.