Information on EC 3.4.22.25 - glycyl endopeptidase

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The expected taxonomic range for this enzyme is: Carica papaya

EC NUMBER
COMMENTARY hide
3.4.22.25
-
RECOMMENDED NAME
GeneOntology No.
glycyl endopeptidase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
preferential cleavage: Gly-/-, in proteins and small molecule substrates
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
CAS REGISTRY NUMBER
COMMENTARY hide
149719-24-4
-
92228-52-9
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Papaya latex
-
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
aggrecan + H2O
?
show the reaction diagram
-
-
-
-
-
azocasein + H2O
trichloroacetic acid-soluble peptides from azocasein
show the reaction diagram
-
10% of the activity of papain
-
-
benzyloxycarbonyl-Gly-Leu-NH + H2O
L-leucine amide + N-benzyloxycarbonyl-glycine
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Gly-Phe-NH + H2O
L-phenylalanine amide + N-benzyloxycarbonyl-glycine
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Gly-Tyr-NH + H2O
L-tyrosine amide + N-benzyloxycarbonyl-glycine
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Gly-Tyr-Oet + H2O
L-tyrosine ethyl ester + N-benzyloxycarbonyl-glycine
show the reaction diagram
-
-
-
?
casein + H2O
?
show the reaction diagram
Papaya latex
-
-
-
-
?
casein + H2O
trichloroacetic acid-soluble peptides derived from casein
show the reaction diagram
-
10% of the activity of papain
-
-
Chicken cystatin + H2O
?
show the reaction diagram
Gly-4-nitroanilide + H2O
glycine + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
Hemoglobin + H2O
Trichloroacetic acid-soluble peptides from haemoglobin
show the reaction diagram
-
-
-
-
hide powder azure + H2O
?
show the reaction diagram
-
-
-
-
-
Human cystatin C + H2O
?
show the reaction diagram
N-Acetyl-L-Phe-Gly 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
N-Benzyloxycarbonyl-Gly 4-nitrophenyl ester + H2O
?
show the reaction diagram
-
-
-
-
-
protein + H2O
peptides
show the reaction diagram
tert-Butyloxycarbonyl-Ala-Ala-Ala 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
-
-
tert-Butyloxycarbonyl-Ala-Ala-Gly 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
-
-
tert-Butyloxycarbonyl-Ala-Ala-Gly 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
tert-butyloxycarbonyl-Ala-Ala-Gly-p-nitroanilide + H2O
tert-butyloxycarbonyl-Ala-Ala-Gly + p-nitroaniline
show the reaction diagram
-
-
-
?
Trypsin + H2O
Hydrolyzed trypsin
show the reaction diagram
-
16 cleavage points of which 13 are glycyl bonds
-
-
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
protein + H2O
peptides
show the reaction diagram
additional information
?
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Alpha-macroglobulin
-
-
-
E64
-
inactivation
iodoacetamide
iodoacetate
L-3-carboxy-2,3-epoxypropionylleucylamido(4-guanidino)butane
-
-
papaya cystatin
-
-
-
Peptide aldehydes
-
despite the presence of bulky sidechains in P1
Peptidyl diazomethanes
Pro-peptide
-
the enzyme is inhibited by its own pro region after cleavage, also pro-peptide of other C1 peptidase family enzymes can be inhibitory
-
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
the enzyme depends on a thiolate ion for activity
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.08
tert-butyloxycarbonyl-Ala-Ala-Ala 4-methylcoumarin 7-amide
-
-
0.16
tert-butyloxycarbonyl-Ala-Ala-Gly 4-methylcoumarin 7-amide
-
-
5.2
tert-butyloxycarbonyl-Ala-Ala-Gly 4-nitroanilide
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.08
tert-butyloxycarbonyl-Ala-Ala-Ala 4-methylcoumarin 7-amide
Carica papaya
-
-
5
tert-butyloxycarbonyl-Ala-Ala-Gly 4-methylcoumarin 7-amide
Carica papaya
-
-
22
tert-butyloxycarbonyl-Ala-Ala-Gly 4-nitroanilide
Carica papaya
-
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0000034
papapya cystatin
-
-
-
0.00086
Pro-peptide
-
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 7.5
-
tert-butyloxycarbonyl-Ala-Ala-Gly 4-nitroanilide
7.5
Papaya latex
-
assay at
8
-
assay at
additional information
-
pI: 10.6-10.7
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 9.5
-
3.0: 15% of maximal activity, 9.5: about 75% of maximal activity, tert-butyloxycarbonyl-Ala-Ala-Gly 4-nitroanilide
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
Papaya latex
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
unripe
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
23000
Papaya latex
-
SDS-PAGE
23310
-
Carica papaya, calculation from amino acid sequence
24000
-
Carica papaya, sedimentation analysis, meniscus depletion method
additional information
-
three-dimensional structure
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
-
1 * 23313
additional information
-
three-dimensional structure comparison to other latex peptidase
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
no glycoprotein
proteolytic modification
-
the enzyme is synthesized as preproenzyme
additional information
-
enzyme is synthesized as inactive proenzyme, and rapidly converted to the active from within 2 min after wounding of the plant
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
the enzyme undergoes a conformational transition at low pH and 37°C, that instantaneously and irreversibly converts the native form into molten globules, which are unstable and rapidly degraded by pepsin
667431
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
recombinant proregion, 60 min, stable
additional information
-
factors effecting thermostability
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation
different methods, overview
-
glycyl endopeptidase from papaya latex is partitioned using aqueous two-phase (10% PEG 6000- 10% (NH4)2SO4) in combination with NH4(SO4)2 precipitation.The partially purified glycyl endopeptidase shows the potential in production of antioxidative gelatin hydrolysates. The enzyme fraction contain lower odorous compounds in papaya latex. The gelatin hydrolysate produced using the selected fraction have negligible odorous compounds
Papaya latex
-
native enzyme from latex by ion exchange and adsorption chromatography, and gel filtration
-
several ligands possible for affinity chromatography are presented, fractionation on hydrophobic and cation-exchange supports, overview
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression of the pro-region of papaya proteinase IV in Escherichia coli
-
expression of the proregion of the enzyme in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
G27E
-
site-directed mutagenesis, high reduced activity with substrate having an Arg at P1 position
G73R
-
site-directed mutagenesis, high reduced activity with substrate having an Arg at P1 position, decreased affinity for cystatin C
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
-
plant protease proregions have a potential as regulators of cysteine proteinases in biotechnological systems and to target proteases of pests
nutrition
-
the enzyme needs to be protected from acid denaturation and proteolysis in the gut after oral administration to be effective as cysteine protease