Information on EC 3.4.22.30 - Caricain

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The expected taxonomic range for this enzyme is: Carica papaya

EC NUMBER
COMMENTARY hide
3.4.22.30
-
RECOMMENDED NAME
GeneOntology No.
Caricain
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
hydrolysis of proteins with broad specificity for peptide bonds, similar to those of papain and chymopapain
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis
-
-
hydrolysis of peptide bond
CAS REGISTRY NUMBER
COMMENTARY hide
39307-22-7
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Albumin + H2O
?
show the reaction diagram
-
low activity
-
-
-
alpha-N-benzoyl-L-arginine 4-nitroanilide + H2O
alpha-N-benzoyl-L-arginine + 4-nitroaniline
show the reaction diagram
-
-
-
?
azocasein + H2O
?
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Gly 4-nitrophenyl ester + H2O
?
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Lys nitrophenyl ester + H2O
?
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Phe-Arg 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
-
-
Bovine nasal cartilage + H2O
?
show the reaction diagram
-
-
-
-
-
casein + H2O
?
show the reaction diagram
Gliadin + H2O
?
show the reaction diagram
gliadin + H2O
fragments of gliadin
show the reaction diagram
-
-
-
-
?
hide powder azure + H2O
?
show the reaction diagram
-
-
-
-
-
Hippuric acid 4-nitrophenyl ester + H2O
?
show the reaction diagram
-
-
-
-
-
N-acetyl-Phe-Gly methylthionoester + H2O
?
show the reaction diagram
-
synthetic chromogenic substrate, reaction mechanism, computer modeling of intermediate formation
-
?
N-Benzoyl-Arg 4-nitroanilide + H2O
?
show the reaction diagram
N-Benzoyl-Arg amide + H2O
?
show the reaction diagram
-
less active than papain
-
-
-
N-Benzoyl-L-Arg ethyl ester + H2O
?
show the reaction diagram
N-benzoyl-L-arginine ethyl ester + H2O
N-benzoyl-L-arginine + ethanol
show the reaction diagram
-
isoform caricain II, low activity
-
-
?
N-tosyl-L-Arg methyl ester + H2O
N-tosyl-L-Arg + methanol
show the reaction diagram
-
less active than papain
-
-
-
Nalpha-benzoyl-L-Arg-4-nitroanilide + H2O
Nalpha-benzoyl-L-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
protein + H2O
peptides
show the reaction diagram
pyr-Glu-Phe-Leu-4-nitroanilide + H2O
pyr-Glu-Phe-Leu + 4-nitroaniline
show the reaction diagram
-
-
?
Succinyl-albumin + H2O
?
show the reaction diagram
-
-
-
-
-
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
Gliadin + H2O
?
show the reaction diagram
-
-
-
-
?
gliadin + H2O
fragments of gliadin
show the reaction diagram
-
-
-
-
?
protein + H2O
peptides
show the reaction diagram
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Benzyloxycarbonyl-Phe-Arg 4-methylcoumarin 7-amide
-
competitive to succinyl-albumin
Gly-Gly(O-benzyl)Tyr-Arg
-
competitive, affinity is dependent on ionic strength and always lower than that of papain
imidazole
-
-
iodoacetamide
-
-
phenylmethylsulfonyl fluoride
-
weak inhibition
Succinyl-albumin
-
competitive to benzyloxycarbonyl-Phe-Arg 4-methylcoumarin 7-amide
-
trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane
-
i.e. E-64, irreversible, active site-directed
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
thiol group
-
enzyme contains 1 thiol group per molecule, this thiol group is essential for catalytic activity
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
16 - 18
benzoyl-DL-Arg 4-nitroanilide
0.028 - 0.035
benzyloxycarbonyl-Lys nitrophenyl ester
0.0067
Benzyloxycarbonyl-Phe-Arg 4-methylcoumarin 7-amide
-
-
0.009
N-Benzyloxycarbonyl-Gly 4-nitrophenyl ester
-
-
0.000096
Succinyl-albumin
-
-
-
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.14 - 0.15
benzoyl-DL-Arg 4-nitroanilide
49
benzyloxycarbonyl-Lys nitrophenyl ester
Carica papaya
-
pH 5.3 and 5.9
7.1
Benzyloxycarbonyl-Phe-Arg 4-methylcoumarin 7-amide
Carica papaya
-
-
16.7
N-Benzyloxycarbonyl-Gly 4-nitrophenyl ester
Carica papaya
-
-
2.6
Succinyl-albumin
Carica papaya
-
-
-
additional information
additional information
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 9.5
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
assay at
37
-
assay at
additional information
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
23500
-
-
24000
-
Carica papaya, sedimentation equilibrium centrifugation
40000
-
procaricain, fractions are estimated from plots of Kav against molecular weight using dextran blue, human gamma-globulin, bovine serum albumin and trypsin inhibitor as standards, mass spectrometric analysis of trypsin-digested fractions from chromatography are carried out by liquid chromatography/electrospray quadrupole time-of-flight mass spectrometry in positive mode
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 18800, purified from enzyme solution. Protein lacks the N-terminal 41 residues of enzyme, disulfide bond C22-C63 is opened
monomer
-
1 * 24000, Carica papaya, SDS-PAGE
additional information
-
transition state structure
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2 - 3
-
enzyme undergoes conformational transition that instantaneously converts the native form into a molten globule state and is completely irreversible
667431
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
85
-
heat stability of caricain up to 85°C
additional information
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, preparation of the mercury derivative of fully active papaya peptidase A, 50fold molar excess of DTT, 30 days stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
CM Sephadex C-50 column chromatography
-
enzyme is semi-purified with from latex crude extracts by ion exchange chromatography on a CM Sephadex column C-50 and gel filtration
-
recombinant wild-type and mutant enzymes from Escherichia coli, to homogeneity
-
to homogeneity
-
to homogeneity, several ligands possible for affinity chromatography are presented, fractionation on hydrophobic and cation-exchange supports, overview
-
with monoclonal antibodies
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
overexpression of wild-type and mutant enzymes in Escherichia coli BL21(DE3)
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E50A
-
site-directed mutagenesis, reduced activity at pH 6.8
E50A/D158N
-
site-directed mutagenesis, reduced activity at pH 6.8
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant wild-type and mutant enzymes from inclusion bodies after overexpression in scherichia coli
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
-
caricain would be suitable for enzyme therapy in gluten intolerance and appears to have synergistic action with porcine intestinal extracts, potential for enzyme therapy in coeliac disease
food industry
-
the enzyme detoxifies gliadin in wheat dough
nutrition
-
at low pH, enzyme undergoes conformational transition leading to instability and rapid degradation by pepsin. To be effective in gut after oral administration, enzyme needs to be protected against acid denaturation and degradation