Information on EC 3.4.22.6 - chymopapain

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The expected taxonomic range for this enzyme is: Caricaceae

EC NUMBER
COMMENTARY
3.4.22.6
-
RECOMMENDED NAME
GeneOntology No.
chymopapain
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Similar to that of papain
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis
P14080
-
hydrolysis of peptide bond
-
-
hydrolysis of peptide bond
-
-
endopeptidase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Chymopapain
P14080
-
Chymopapain
Q5Q047
-
chymopapain A
-
-
-
-
chymopapain B
-
-
-
-
Chymopapain S
-
-
-
-
papaya proteinase II
-
-
-
-
PPII
-
-
-
-
EC 3.4.4.11
-
-
formerly
-
additional information
-
chymopapain S is chymopapain A
CAS REGISTRY NUMBER
COMMENTARY
9001-09-6
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Albumin + H2O
?
show the reaction diagram
-
-
-
-
?
azocasein + H2O
?
show the reaction diagram
-
-
-
-
?
benzoyl-Gly amide + H2O
benzoyl-Gly + NH3
show the reaction diagram
-
weak activity
-
-
?
benzoyl-L-argininamide + H2O
N-benzoyl-Arg + NH3
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Lys phenyl ester + H2O
benzyloxycarbonyl + phenol
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Phe-Arg-4-methylcoumarin 7-amide + H2O
benzyloxycarbonyl-Phe-Arg-4-methylcoumarin 7-amide
show the reaction diagram
-
-
-
-
?
carbobenzoxy-L-Glu-L-Tyr + H2O
Carbobenzoxy-Glu + Tyr
show the reaction diagram
-
weak activity
-
-
?
casein + H2O
?
show the reaction diagram
-
-
-
-
?
Hemoglobin + H2O
?
show the reaction diagram
-
-
-
-
?
L-Leu amide + H2O
Leu + NH3
show the reaction diagram
-
weak activity
-
-
?
L-Leu-L-Tyr + H2O
Leu + Tyr
show the reaction diagram
-
weak activity
-
-
?
methyl red-Abu-Ala-Pro-Val-Lys-Lys(N5-(5-carboxyfluorescein))-NH2 + H2O
methyl red-Abu-Ala-Pro-Val-Lys + Lys(N5-(5-carboxyfluorescein))-NH2
show the reaction diagram
-
FRET 2, fluorescence resonance energy transfer peptide 2, pH 6.2 or pH 7.4, 10 min, 37C
-
-
?
methyl red-Abu-Ser-Ala-Pro-Val-Lys-Ala-Lys(N5-(5-carboxyfluorescein))-NH2 + H2O
methyl red-Abu-Ser-Ala-Pro-Val-Lys + Ala-Lys(N5-(5-carboxyfluorescein))-NH2
show the reaction diagram
-
pH 6.2 or pH 7.4, 10 min, 37C
-
-
?
N-acetyl-L-Phe-L-Tyr + H2O
?
show the reaction diagram
-
weak activity
-
-
?
N-alpha-benzoyl-DL-arginine-p-nitroanilide + H2O
N-alpha-benzoyl-DL-arginine + p-nitroaniline
show the reaction diagram
-
-
-
-
?
N-alpha-benzoyl-DL-arginine-para-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
N-alpha-benzoyl-L-arginine ethyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
N-alpha-benzoylphenylalanine-arginine-4-methylcoumarin-7-amide + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzoyl-Arg-4-nitroanilide + H2O
N-benzoyl-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
N-benzoyl-Arg-4-nitroanilide + H2O
N-benzoyl-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
?
N-benzoyl-Arg-4-nitroanilide + H2O
N-benzoyl-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
N-benzoyl-L-Arg ethyl ester + H2O
N-benzoyl-Arg + ethanol
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-Gly p-nitrophenyl ester + H2O
N-benzyloxycarbonyl-Gly + p-nitrophenol
show the reaction diagram
-
-
-
-
?
N-tosyl-L-Arg methyl ester + H2O
N-tosyl-Arg + methanol
show the reaction diagram
-
-
-
-
?
succinyl-albumin
?
show the reaction diagram
-
-
-
-
?
succinyl-Phe-Arg-p-nitroanilide + H2O
succinyl-Phe-Arg + p-nitroaniline
show the reaction diagram
-
mechanism of action
-
-
?
methyl red-Abu-Ser-Ala-Pro-Val-Lys-Ala-Lys(N6-(5-carboxyfluorescein))-NH2 + H2O
methyl red-Abu-Ser-Ala-Pro-Val-Lys + Ala-Lys(N6-(5-carboxyfluorescein))-NH2
show the reaction diagram
-
FRET 1, fluorescence resonance energy transfer peptide 1
-
-
?
additional information
?
-
-
the abnormal catalytic site environment of chymopapain A may have consequences for the acylation step of the catalytic act, does not pertub the conformation of the bound acyl group at the acyl-enzyme-intermediate stage of catalysis
-
-
-
additional information
?
-
-
mass spectroscopy studies show unequivocally the specificity of chymopapain toward Ala, Pro, Val and Lys for positions P4 to P1 while not presenting high specificity for residues in position P1
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
additional information
?
-
-
mass spectroscopy studies show unequivocally the specificity of chymopapain toward Ala, Pro, Val and Lys for positions P4 to P1 while not presenting high specificity for residues in position P1
-
-
-
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Albumin
-
inhibits hydrolysis of synthetic substrates
-
Cystatin
-
-
-
Succinyl-albumin
-
inhibits hydrolysis of synthetic substrates
-
iodoacetate
-
-
additional information
-
crystals from potato tubers are predominantly composed of a proteinase inhibitor with a MW of 80000 Da, that inhibits chymopapain
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2,3-Dimercaptopropanol
-
maximal activity in presence of various reducing compounds, e.g. Cys, 2,3-dimercaptopropanol, thioglycolic acid, glutathione
2,3-Dimercaptopropanol
-
accelerates casein digestion
Cys
-
maximal activity in presence of various reducing compounds, e.g. Cys, 2,3-dimercaptopropanol, thioglycolic acid, glutathione
Cys
-
accelerates casein digestion
glutathione
-
maximal activity in presence various reducing compounds, e.g. Cys, 2,3-dimercaptopropanol, thioglycolic acid, glutathione
glutathione
-
accelerates casein digestion
NaN3
-
accelerates casein digestion
thioglycolic acid
-
maximal activity in presence of various reducing compounds, e.g. Cys, 2,3-dimercaptopropanol, thioglycolic acid, glutathione
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
5
-
benzoyl-Arg-4-nitroanilide
-
pH 7.4
8.6
-
benzoyl-Arg-4-nitroanilide
-
pH 4.0
13
-
benzoyl-Arg-4-nitroanilide
-
pH 9.2
0.0075
-
benzyloxycarbonyl-Lys phenyl ester
-
at pH 6.0
0.01
-
benzyloxycarbonyl-Lys phenyl ester
-
at pH 6.9 and at pH 4.9
0.0025
-
benzyloxycarbonyl-Phe-Arg-4-methylcoumarin 7-amide
-
-
0.0502
-
methyl red-Abu-Ala-Pro-Val-Lys-Lys(N5-(5-carboxyfluorescein))-NH2
-
at pH 7.4, in 100 mM phosphate-buffered saline; pH 7.4
0.0506
-
methyl red-Abu-Ala-Pro-Val-Lys-Lys(N5-(5-carboxyfluorescein))-NH2
-
at pH 6.2, in 10 mM sodium acetate, 0.1 mM EDTA and 1 mM cysteine; pH 6.2
0.023
-
methyl red-Abu-Ser-Ala-Pro-Val-Lys-Ala-Lys(N6-(5-carboxyfluorescein))-NH2
-
at pH 6.2, in 10 mM sodium acetate, 0.1 mM EDTA and 1 mM cysteine; pH 6.2
0.026
-
methyl red-Abu-Ser-Ala-Pro-Val-Lys-Ala-Lys(N6-(5-carboxyfluorescein))-NH2
-
at pH 7.4, in 100 mM phosphate-buffered saline; pH 7.4
0.000131
-
Succinyl-albumin
-
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.003
-
benzoyl-Arg-4-nitroanilide
-
pH 4.0 and pH 7.4
0.05
-
benzoyl-Arg-4-nitroanilide
-
pH 9.2
14
-
benzyloxycarbonyl-Lys phenyl ester
-
at pH 4.9
15.5
-
benzyloxycarbonyl-Lys phenyl ester
-
at pH 6.0 and at pH 6.9
2.9
-
benzyloxycarbonyl-Phe-Arg-4-methylcoumarin 7-amide
-
-
246
-
methyl red-Abu-Ala-Pro-Val-Lys-Lys(N5-(5-carboxyfluorescein))-NH2
-
at pH 6.2, in 10 mM sodium acetate, 0.1 mM EDTA and 1 mM cysteine
590
-
methyl red-Abu-Ala-Pro-Val-Lys-Lys(N5-(5-carboxyfluorescein))-NH2
-
at pH 7.4, in 100 mM phosphate-buffered saline
133
-
methyl red-Abu-Ser-Ala-Pro-Val-Lys-Ala-Lys(N6-(5-carboxyfluorescein))-NH2
-
at pH 6.2, in 10 mM sodium acetate, 0.1 mM EDTA and 1 mM cysteine
360
-
methyl red-Abu-Ser-Ala-Pro-Val-Lys-Ala-Lys(N6-(5-carboxyfluorescein))-NH2
-
at pH 7.4, in 100 mM phosphate-buffered saline
4
-
Succinyl-albumin
-
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.154
-
-
-
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.5
8
-
benzoyl-L-argininamide
6.2
-
-
assay at
7
9
-
casein digestion
7
-
-
hemoglobin digestion
7.2
-
P14080
enzyme is dissolved in phosphate buffer and used with a suspension of lysosomes in 0.154 mol/l sodium chloride
7.2
-
-
casein or hemoglobin digestion
7.4
-
-
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6
9.8
-
about 65% of maximal activity at pH 6.0 and at pH 9.8
6.2
7.4
-
10 min, 37C
additional information
-
-
chymopapain has a broad working pH range
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
assay at; pH 6.2 or pH 7.4, 10 min
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
latex from ripe and unripe fruit. Repeated wounding results in either accumulation or activation of enzyme as well as papain, caricain and more cysteine proteases
Manually annotated by BRENDA team
-
repeated wounding results in either, accumulation or activation (or both of them) of chymopapain
Manually annotated by BRENDA team
P14080
; estimated as a fraction of papaya latex
Manually annotated by BRENDA team
-
of unripe fruit
Manually annotated by BRENDA team
Vasconcellea cundinamarcensis Hook
-
-
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
24000
-
-
equilibrium sedimentation
24100
-
-
enzyme form III, gel filtration
24400
-
-
enzyme form II and enzyme form IV, gel filtration
34500
-
-
chymopapain B, low speed sedimentation
36000
-
P14080
fraction is 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
36400
-
-
chymopapain A, low speed sedimentation
additional information
-
-
complete primary structure
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 24700, chymopapain S, SDS-PAGE
monomer
-
x * 24000, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
proteolytic modification
-
the enzyme is synthesized as inactive precursor that converts into mature enzyme within 2 min after wounding the plant when the latex is abruptly expelled
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
1
-
-
at low pH, the enzyme undergoes a conformational transition that instantaneously converts their native form into molten globule that are quite unstable and rapidly degraded by pepsin. The denatured state of these proteinase which results from acid treatment is completely irreversible
1.5
-
-
protein unfolds following biphasic kinetics. Two different effects of electrolyte concentration on unfolding reaction are observed. At low ionic strength, the ionic atmosphere causes an increase in reaction rates, regardless of the type of ions being present. This effect is attributed to a general electrostatic screening of charge-charge interactions in the macromolecule. At high ionic strength, each electrolyte exerts a distinctly different effect: both rate constants are largely increased by guanidinium-HCl but slightly by LiCl. Na2SO4 decreases the value of both unfolding rates
2
3
-
enzyme undergoes conformational transition that instantaneously converts the native form into a molten globule state and is completely irreversible
2
-
-
stable
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
pH 5, 72 h, about 35% loss of its activity
75
-
-
half-life at pH 2.1: 2 min, half-life at pH 7.2: 75 min
additional information
-
-
irreversible thermal denaturation according to a first order reaction in which the macromolecule is globally unfolded
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
during purification, the thiol function of the Cys residues are protected either as mixed disulfides with cysteamine or 2-thiopyridone or as S-sulfenylthiosulfate derivative or after blocking with PCMB
-
freeze-drying causes 7% loss of activity
-
mercury prevents both irreversible oxidation and autocatalysis
-
PEG 20000 destabilizes chymopapain
-
OXIDATION STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
mercury prevents irreversible oxidation
-
81571
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, freeze-dryed, stable for at least 1 month
-
0C, pH 2.0, stable
-
4C, 15% loss of activity after 2 weeks
-
4C, stable in solution for at least 1 month
-
frozen, 50% loss of activity after 24 h
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
; enzyme is semi-purified with from latex crude extracts by ion exchange chromatography on a CM Sephadex column C-50 and gel filtration
P14080
addition of polyethyleneglycol facilitates purification
-
chymopapain B
-
chymopapain S
-
commercial preparation
-
supports: phenyl-Hg+ CH3COO- and glutathione-S-S-(2-pyridyl), further purification through the S-thiol poly(ethylene glycol) derivatization technique
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
medicine
-
significantly more nucleus can be removed from a disc using a bilateral rather than a unilateral approach. A unilateral approach using rongeurs alone leads to removal of only about one-third of the nucleus. Rongeurs, followed by chymopapain, lead to removal of significantly more nucleus than when rongeurs are used alone. The most effective method is a bilateral approach using rongeurs followed by chymopapain where about three-quarters of the nucleus can be removed. This approach can create sufficient space for the implant insertion. A brush, which can be inserted through a trocar, assists in the removal of strands of nucleus that are loosened by chymopapain. Experiments are carried out on sheep spines since they are increasingly being used as models for human spines
medicine
-
enzyme is used for a number of biomedical applications; used for pain relief in treatment of herniated invertebral disc
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
medicine
-
is used for chemonucleolysis of damaged human intervertebral spinal discs
medicine
Vasconcellea cundinamarcensis Hook
-
is used for chemonucleolysis of damaged human intervertebral spinal discs
-