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Information on EC 3.4.99.B2 - D-aspartyl endopeptidase
for references in articles please use BRENDA:EC3.4.99.B2
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Old data from external sources may still be available.
Please check here for current data: EC unknown
Please check here for current data: EC unknown
preliminary BRENDA-supplied EC number
EC Tree 3 Hydrolases
3.4 Acting on peptide bonds (peptidases)
3.4.99 Endopeptidases of unknown catalytic mechanism (sub-subclass is currently empty)
3.4.99.B2 D-aspartyl endopeptidase
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
- 3.4.99.B2
- alzheimer
-
cataracts
-
noxious
- d-aspartate
-
misfolding
- proteasome
-
aging-related
-
arteriosclerosis
-
lactacystin-sensitive
-
multi-complex
-
racemized
- prion
- paenibacillus
-
d-isomers
- lactacystin
- elegans
- analysis
Reaction Schemes
showpaenidases specifically recognize internal D-Asp residues and hydrolyze them on the COOH side
Synonyms
paenidase, d-aspartyl endopeptidase, more
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
D-aspartyl endopeptidase
DAEP
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
paenidases specifically recognize internal D-Asp residues and hydrolyze them on the COOH side
-
-
-
-
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
D-Asp-L-Ala-L-Glu-L-Phe-L-Arg-L-His-D-Asp-L-Ser-Gly-L-Tyr + H2O
?
-
Substrates: -
Products: -
Products: -
?
DAEFRH-(D-Asp)-GSY + H2O
DAEFRH-(D-Asp) + GSY
-
Substrates: synthetic Abeta peptide. No cleavage of the D-Aspcontaining Abeta peptide, paenidase I and II. Paenidases specifically recognize internal D-Asp residues and hydrolyze them on the COOH side
Products: -
Products: -
?
L-Thr-L-Val-L-Leu-D-alpha-Asp-L-Ser-Gly-L-Ile-L-Ser-L-Glu-L-Val-L-Arg + H2O
?
-
Substrates: -
Products: -
Products: -
?
succinyl-D-Asp-alpha-4-methylcoumaryl-7-amide + H2O
succinyl-D-Asp + 7-amino-4-methylcoumarin
-
Substrates: -
Products: -
Products: -
?
[D-Asp]AEFRH[D-Asp]SGY + H2O
[D-Asp]AEFRHX + SGY
-
Substrates: the D-Abeta1-10 peptide is degraded at D-Asp7 and two peptides are produced [D-Asp]AEFRHX and SGY. X is not identified in this study. L-Abeta110 peptide could not be degraded by this enzyme
Products: -
Products: -
?
[D-Asp]AEFRH[D-Asp]SGY + H2O
[D-Asp]AEFRH[D-Asp] + SGY
-
Substrates: the D-Abeta1-10 peptide is degraded at D-Asp7 and two peptides are produced [D-Asp]AEFRHX and SGY. X is not identified in this study. L-Abeta110 peptide could not be degraded by this enzyme
Products: -
Products: -
?
succinyl-D-Asp-4-methylcoumaryl-7-amide + H2O
succinyl-D-Asp + 7-amino-4-methylcoumarin
-
Substrates: -
Products: -
Products: -
?
succinyl-D-Asp-4-methylcoumaryl-7-amide + H2O
succinyl-D-Asp + 7-amino-4-methylcoumarin
-
Substrates: paenidases specifically recognize internal D-Asp residues and hydrolyze them on the COOH side
Products: -
Products: -
?
succinyl-D-Asp-7-amido-4-methylcoumarin + H2O
succinyl-D-Asp + 7-amino-4-methylcoumarin
-
Substrates: -
Products: -
Products: -
?
succinyl-D-Asp-7-amido-4-methylcoumarin + H2O
succinyl-D-Asp + 7-amino-4-methylcoumarin
Substrates: -
Products: -
Products: -
?
succinyl-D-Asp-p-nitroanilide + H2O
succinyl-D-Asp + p-nitroaniline
-
Substrates: paenidases specifically recognize internal D-Asp residues and hydrolyze them on the COOH side
Products: -
Products: -
?
succinyl-D-Asp-p-nitroanilide + H2O
succinyl-D-Asp + p-nitroaniline
Substrates: -
Products: -
Products: -
?
additional information
?
-
-
Substrates: the enzyme may serve as a scavenger against accumulation of racemized proteins in aging
Products: -
Products: -
?
additional information
?
-
-
Substrates: no activity with succinyl-L-Asp-7-amido-4-methylcoumarin, L-Thr-L-Val-L-Leu-L-alpha-Asp-L-Ser-Gly-L-Ile-L-Ser-L-Glu-L-Val-L-Arg, L-Thr-L-Val-L-Leu-D-beta-Asp-L-Ser-Gly-L-Ile-L-Ser-L-Glu-L-Val-L-Arg and L-Thr-L-Val-L-Leu-D-beta-Asp-L-Ser-Gly-L-Ile-L-Ser-L-Glu-L-Val-L-Arg
Products: -
Products: -
?
additional information
?
-
-
Substrates: the enzyme may serve as a scavenger against accumulation of racemized proteins in aging
Products: -
Products: -
?
additional information
?
-
-
Substrates: the accumulation of D-isomers of aspartic acid in proteins during aging is involved in the pathogenesis of Alzheimers disease, cataracts, and arteriosclerosis. The enzyme may serve as a scavenger against accumulation of racemized proteins in aging
Products: -
Products: -
?
additional information
?
-
-
Substrates: no activity with acetyl-Tyr-Val-Ala-Asp-4-methylcoumaryl-7-amide, acetyl-Asp-Glu-Val-Asp-4-methylcoumaryl-7-amide, succinyl-Gly-Pro-Leu-Gly-Pro-4-methylcoumaryl-7-amide, benzyloxycarbonyl-Phe-Arg-4-methylcoumaryl-7-amide, benzyloxycarbonyl-Arg-Arg-4-methylcoumaryl-7-amide, Leu-4-methylcoumaryl-7-amide, succinyl-Leu-Leu-Val-Tyr-4-methylcoumaryl-7-amide, benzyloxycarbonyl-Leu-Leu-Glu-4-methylcoumaryl-7-amide, Ala-Ala-Phe-4-methylcoumaryl-7-amide, succinyl-Ala-[D-Ser]-4-methylcoumaryl-7-amide, succinyl-[D-Ser]-4-methylcoumaryl-7-amide
Products: -
Products: -
?
additional information
?
-
-
Substrates: no activity with acetyl-Tyr-Val-Ala-Asp-4-methylcoumaryl-7-amide, acetyl-Asp-Glu-Val-Asp-4-methylcoumaryl-7-amide, succinyl-Gly-Pro-Leu-Gly-Pro-4-methylcoumaryl-7-amide, benzyloxycarbonyl-Phe-Arg-4-methylcoumaryl-7-amide, benzyloxycarbonyl-Arg-Arg-4-methylcoumaryl-7-amide, Leu-4-methylcoumaryl-7-amide, succinyl-Leu-Leu-Val-Tyr-4-methylcoumaryl-7-amide, benzyloxycarbonyl-Leu-Leu-Glu-4-methylcoumaryl-7-amide, Ala-Ala-Phe-4-methylcoumaryl-7-amide, succinyl-Ala-[D-Ser]-4-methylcoumaryl-7-amide, succinyl-[D-Ser]-4-methylcoumaryl-7-amide
Products: -
Products: -
?
additional information
?
-
-
Substrates: 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
Products: -
Products: -
?
additional information
?
-
Substrates: enzyme specifically recognizes an internal D-Asp residue to cleave [D-Asp]-X peptide bonds
Products: -
Products: -
?
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
additional information
?
-
-
Substrates: the enzyme may serve as a scavenger against accumulation of racemized proteins in aging
Products: -
Products: -
?
additional information
?
-
-
Substrates: the enzyme may serve as a scavenger against accumulation of racemized proteins in aging
Products: -
Products: -
?
additional information
?
-
-
Substrates: the accumulation of D-isomers of aspartic acid in proteins during aging is involved in the pathogenesis of Alzheimers disease, cataracts, and arteriosclerosis. The enzyme may serve as a scavenger against accumulation of racemized proteins in aging
Products: -
Products: -
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
benzoyl-Arg-His-D-Asp-CH2Cl
-
1 mM, about 70% inhibition of paenidase I and II
Co2+
-
1.0 mM, 31% inhibition of paenidase I, 46% inhibition of paenidase II
Mn2+
-
1.0 mM MnCl2, 31% inhibition of paenidase I, 60% inhibition of paenidase II
additional information
-
no inhibition: N-p-tosyl-L-phenylalanine chloromethyl ketone, N-p-tosyl-L-lysine chloromethyl ketone, phenylmethylsulfonyl fluoride, trans-epoxysuccinyl-L-leucylamido(4-guanidino)butane, L-alanyl-L-alanyl-L-phenylalanyl-chloromethane, chymostatin, EDTA, epoxomicin, N-acetyl-L-leucyl-L-leucyl-norleucinal
-
additional information
-
no inhibition: N-p-tosyl-L-phenylalanine chloromethyl ketone, N-p-tosyl-L-lysine chloromethyl ketone, phenylmethylsulfonyl fluoride, trans-epoxysuccinyl-L-leucylamido(4-guanidino)butane, L-alanyl-L-alanyl-L-phenylalanyl-chloromethane, chymostatin, EDTA
-
additional information
-
no inhibition: iodoacetic acid, p-chloromercuribenzoate, EDTA, 1,10-phenanthroline, amastatin, antipain, diisopropylfluorophosphate, soybean trypsin inhibitor
-
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DISEASE
TITLE OF PUBLICATION
LINK TO PUBMED
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.12
succinyl-D-Asp-alpha-4-methylcoumaryl-7-amide
-
pH 8,5, 37°C
0.14
succinyl-D-Asp-7-amido-4-methylcoumarin
mutant H276A, pH 7.0, 37°C
0.97
succinyl-D-Asp-7-amido-4-methylcoumarin
wild-type, pH 7.0, 37°C
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.34
succinyl-D-Asp-7-amido-4-methylcoumarin
mutant H276A, pH 7.0, 37°C
2.88
succinyl-D-Asp-7-amido-4-methylcoumarin
wild-type, pH 7.0, 37°C
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2.37
succinyl-D-Asp-7-amido-4-methylcoumarin
mutant H276A, pH 7.0, 37°C
2.97
succinyl-D-Asp-7-amido-4-methylcoumarin
wild-type, pH 7.0, 37°C
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IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
8
-
paenidases I and II, succinyl-D-Asp-4-methylcoumaryl-7-amide or succinyl-D-Asp-p-nitroanilide as a substrate
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Highest Expressing Human Cell Lines
Cell Line LinksPlease wait a moment until the data is sorted. This message will disappear when the data is sorted.
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
33000
-
1 * 33000, paenidase II lacks 5 of the residues present in the N-terminal sequence of paenidase I, paenidase II
34170
-
paenidase II, paenidase II lacks 5 of the residues present in the N-terminal sequence of paenidase I, MALDI-TOF/MS
600000
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
x * 32000, mature recombinant protein, x * 55000, recombinant pro-protein, SDS-PAGE, x * 55756, pro-orotein, calculated from sequence
monomer
monomer
-
1 * 33000, paenidase II lacks 5 of the residues present in the N-terminal sequence of paenidase I, paenidase II
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30
40
-
pH 8, 30 min, paenidase I: about 20% loss of activity, paenidase II: about 30% loss of activity
60
-
pH 8, 30 min, paenidase I: about 80% loss of activity, paenidase II: about 90% loss of activity
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
-
a system for screening D-Asp-containing proteins is developed by using D-aspartyl endopeptidase and a two-dimensional gel electrophoresis
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Kinouchi, T.; Ishiura, S.; Mabuchi, Y.; Urakami-Manaka, Y.; Nishio, H.; Nishiuchi, Y.; Tsunemi, M.; Takada, K.; Watanabe, M.; Ikeda, M.; Matsui, H.; Tomioka, S.; Kawahara, H.; Hamamoto, T.; Suzuki, K.; Kagawa, Y.
Mammalian D-aspartyl endopeptidase: a scavenger for noxious racemized proteins in aging
Biochem. Biophys. Res. Commun.
314
730-736
2004
Mus musculus, no activity in Caenorhabditis elegans, no activity in Escherichia coli, no activity in Saccharomyces cerevisiae, Oryctolagus cuniculus
brenda
Kinouchi, T.; Nishio, H.; Nishiuchi, Y.; Tsunemi, M.; Takada, K.; Hamamoto, T.; Kagawa, Y.; Fujii, N.
Isolation and characterization of mammalian D-aspartyl endopeptidase
Amino Acids
32
79-85
2007
Mus musculus, Oryctolagus cuniculus
brenda
Sakai-Kato, K.; Kinouchi, T.; Fujii, N.; Imai, K.; Utsunomiya-Tate, N.
Screening system for D-Asp-containing proteins using D-aspartyl endopeptidase and two-dimensional gel electrophoresis
Amino Acids
36
125-129
2009
Oryctolagus cuniculus
brenda
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
J. Biochem.
139
197-202
2006
Paenibacillus sp.
brenda
Kinouchi, T.; Fujii, N.
Structural consideration of mammalian D-aspartyl endopeptidase
Chem. Biodivers.
7
1403-1407
2010
Mus musculus
brenda
Kinouchi, T.; Fujii, N.; Fujii, N.
Substrate stereoselectivity of mammalian D-aspartyl endopeptidase
J. Chromatogr. B
879
3349-3352
2011
Mus musculus
brenda
Nirasawa, S.; Nakahara, K.; Takahashi, S.
Cloning and characterization of the novel D-aspartyl endopeptidase, paenidase, from Paenibacillus sp. B38
J. Biochem.
164
103-112
2018
Paenibacillus sp. B38 (A0A2Z6BCG6)
brenda
EXTERNAL LINKS (specific for EC-Number 3.4.99.B2)
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