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Literature summary for 3.4.99.B2 extracted from

  • Takahashi, S.; Ogasawara, H.; Hiwatashi, K.; Hori, K.; Hata, K.; Tachibana, T.; Itoh, Y.; Sugiyama, T.
    Paenidase, a novel D-aspartyl endopeptidase from Paenibacillus sp. B38: purification and substrate specificity (2006), J. Biochem., 139, 197-202.
    View publication on PubMed

Inhibitors

Inhibitors Comment Organism Structure
benzoyl-Arg-His-D-Asp-CH2Cl 1 mM, about 70% inhibition of paenidase I and II Paenibacillus sp.
Co2+ 1.0 mM, 31% inhibition of paenidase I, 46% inhibition of paenidase II Paenibacillus sp.
Mn2+ 1.0 mM MnCl2, 31% inhibition of paenidase I, 60% inhibition of paenidase II Paenibacillus sp.
additional information no inhibition: iodoacetic acid, p-chloromercuribenzoate, EDTA, 1,10-phenanthroline, amastatin, antipain, diisopropylfluorophosphate, soybean trypsin inhibitor Paenibacillus sp.
NEM 0.1 mM, about 10% inhibition Paenibacillus sp.
pepstatin 0.01 mg/ml, about 50% inhibition of paenidase I and II Paenibacillus sp.
Zn2+ 0.25 mM ZnCl2, 98% inhibition of paenidase I and II Paenibacillus sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1
-
succinyl-D-Asp-4-methylcoumaryl-7-amide pH 8, 37°C, paenidase II Paenibacillus sp.
1.03
-
succinyl-D-Asp-p-nitroanilide pH 8, 37°C, paenidase I Paenibacillus sp.
1.25
-
succinyl-D-Asp-4-methylcoumaryl-7-amide pH 8, 37°C, paenidase I Paenibacillus sp.
1.26
-
succinyl-D-Asp-p-nitroanilide pH 8, 37°C, paenidase II Paenibacillus sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
33000
-
1 * 33000, paenidase II lacks 5 of the residues present in the N-terminal sequence of paenidase I, paenidase II Paenibacillus sp.
34000
-
1 * 34000, paenidase I Paenibacillus sp.
34170
-
paenidase II, paenidase II lacks 5 of the residues present in the N-terminal sequence of paenidase I, MALDI-TOF/MS Paenibacillus sp.
34790
-
paenidase I, MALDI-TOF/MS Paenibacillus sp.
35000
-
gel filtration Paenibacillus sp.

Organism

Organism UniProt Comment Textmining
Paenibacillus sp.
-
-
-

Purification (Commentary)

Purification (Comment) Organism
paenidases I and II Paenibacillus sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2370
-
paenidase II Paenibacillus sp.
2880
-
paenidase I Paenibacillus sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
DAEFRH-(D-Asp)-GSY + H2O synthetic Abeta peptide. No cleavage of the D-Asp–containing Abeta peptide, paenidase I and II. Paenidases specifically recognize internal D-Asp residues and hydrolyze them on the COOH side Paenibacillus sp. DAEFRH-(D-Asp) + GSY
-
?
additional information no activity with: acetyl-L-Asp-p-nitroanilide, D-Asp-p-nitroanilide, A-Ala-p-nitroanilide, D-Leu-p-nitroanilide, D-Phe-p-nitroanilide, L-Asp-p-nitroanilide, L-Ala-p-nitroanilide, L-Leu-p-nitroanilide, L-Phe-p-nitroanilide, L-Arg-p-nitroanilide, L-Glu-p-nitroanilide, Gly-p-nitroanilide, L-His-p-nitroanilide, L-Ile-p-nitroanilide, L-Lys-p-nitroanilide, L-Met-p-nitroanilide, L-Pro-p-nitroanilide, L-Val-p-nitroanilide, p-nitroanilide, succinyl-L-Ala-L-Pro-L-Ala, succinyl-L-Ala-L-Ala-L-Ala-p-nitroanilide, L-Pyr-p-nitroanilide, Arg-4-methylcoumaryl-7-amide, benzoyl-Arg-4-methylcoumaryl-7-amide, tert-butyloxycarbonyl-Gln-Ala-Arg-4-methylcoumaryl-7-amide, Pro-Phe-Arg-4-methylcoumaryl-7-amide, acetyl-Tyr-val-Ala-Asp-4-methylcoumaryl-7-amide, acetyl-ASp-GLu-Val-Asp-4-methylcoumaryl-7-amide, acetyl-Val-Glu-Ile-Asp-4-methylcoumaryl-7-amide Paenibacillus sp. ?
-
?
succinyl-D-Asp-4-methylcoumaryl-7-amide + H2O paenidases specifically recognize internal D-Asp residues and hydrolyze them on the COOH side Paenibacillus sp. succinyl-D-Asp + 7-amino-4-methylcoumarin
-
?
succinyl-D-Asp-p-nitroanilide + H2O paenidases specifically recognize internal D-Asp residues and hydrolyze them on the COOH side Paenibacillus sp. succinyl-D-Asp + p-nitroaniline
-
?

Subunits

Subunits Comment Organism
monomer 1 * 33000, paenidase II lacks 5 of the residues present in the N-terminal sequence of paenidase I, paenidase II Paenibacillus sp.
monomer 1 * 34000, paenidase I Paenibacillus sp.

Synonyms

Synonyms Comment Organism
DAEP
-
Paenibacillus sp.
paenidase
-
Paenibacillus sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30
-
pH 8, 30 min, stable below, paenidases I and II Paenibacillus sp.
40
-
pH 8, 30 min, paenidase I: about 20% loss of activity, paenidase II: about 30% loss of activity Paenibacillus sp.
50
-
pH 8, 30 min, about 40% loss of activity, paenidases I and II Paenibacillus sp.
60
-
pH 8, 30 min, paenidase I: about 80% loss of activity, paenidase II: about 90% loss of activity Paenibacillus sp.
70
-
pH 8, 30 min, complete loss of activity, paenidases I and II Paenibacillus sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.66
-
succinyl-D-Asp-4-methylcoumaryl-7-amide pH 8, 37°C, paenidase II Paenibacillus sp.
1.93
-
succinyl-D-Asp-4-methylcoumaryl-7-amide pH 8, 37°C, paenidase I Paenibacillus sp.
4.7
-
succinyl-D-Asp-p-nitroanilide pH 8, 37°C, paenidase I Paenibacillus sp.
6.6
-
succinyl-D-Asp-p-nitroanilide pH 8, 37°C, paenidase II Paenibacillus sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
paenidases I and II, succinyl-D-Asp-4-methylcoumaryl-7-amide or succinyl-D-Asp-p-nitroanilide as a substrate Paenibacillus sp.

pH Range

pH Minimum pH Maximum Comment Organism
7 9 pH 7: about 80% of maximal activity, pH 9.0: about 60% of maximal activity, paenidases I and II, succinyl-D-Asp-4-methylcoumaryl-7-amide as substrate Paenibacillus sp.