Information on EC 3.4.22.6 - chymopapain

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The expected taxonomic range for this enzyme is: Caricaceae

EC NUMBER
COMMENTARY hide
3.4.22.6
-
RECOMMENDED NAME
GeneOntology No.
chymopapain
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Similar to that of papain
show the reaction diagram
-
-
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis
-
-
hydrolysis of peptide bond
CAS REGISTRY NUMBER
COMMENTARY hide
9001-09-6
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2,4-dichlorocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 43%, 80:20 anti/syn, ee (anti): 81%
-
-
?
2,6-dichlorocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 39%, 99:01 anti/syn, ee (anti): 58%
-
-
?
2-chlorocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 30%, 86:14 anti/syn, ee (anti): 79%
-
-
?
2-furanyl + cyclohexanone
?
show the reaction diagram
-
yield: 16%, 63:37 anti/syn, ee (anti): 61%
-
-
?
2-nitrocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 24%, 87:13 anti/syn, ee (anti): 82%
-
-
?
3-chlorocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 32%, 65:35 anti/syn, ee (anti): 93%
-
-
?
3-nitrocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 41%, 87:13 anti/syn, ee (anti): 86%
-
-
?
4-bromocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 29%, 81:19 anti/syn, ee (anti): 84%
-
-
?
4-chlorocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 47%, 69:31 anti/syn, ee (anti): 75%
-
-
?
4-cyanobenzaldehyde + cyclohexanone
4-[hydroxy(2-oxocyclohexyl)methyl]benzonitrile
show the reaction diagram
4-cyanoclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 60%, 72:28 anti/syn, ee (anti): 76%
-
-
?
4-methylcyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 23%, 63:37 anti/syn, ee (anti): 96%
-
-
?
4-nitrocyclohexan-1-one + acetaldehyde
?
show the reaction diagram
-
yield: 12%, ee (anti): 14%
-
-
?
4-nitrocyclohexan-1-one + cycloheptanone
?
show the reaction diagram
-
yield: 19%, 47:53 anti/syn, ee (anti): 26%
-
-
?
4-nitrocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 69%, 63:37 anti/syn, ee (anti): 78%
-
-
?
4-nitrocyclohexan-1-one + cyclopentanone
?
show the reaction diagram
-
yield: 73%, 56:44 anti/syn, ee (anti): 52%
-
-
?
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
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, 37°C
-
-
?
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
-
-
?
N,N-diBoc-dityrosine-(isoniacid)2 + H2O
N,N-diBoc-dityrosine + 2 isoniacid
show the reaction diagram
-
-
-
-
?
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-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
-
-
?
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
2,4-dichlorocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 43%, 80:20 anti/syn, ee (anti): 81%
-
-
?
2,6-dichlorocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 39%, 99:01 anti/syn, ee (anti): 58%
-
-
?
2-chlorocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 30%, 86:14 anti/syn, ee (anti): 79%
-
-
?
2-furanyl + cyclohexanone
?
show the reaction diagram
-
yield: 16%, 63:37 anti/syn, ee (anti): 61%
-
-
?
2-nitrocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 24%, 87:13 anti/syn, ee (anti): 82%
-
-
?
3-chlorocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 32%, 65:35 anti/syn, ee (anti): 93%
-
-
?
3-nitrocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 41%, 87:13 anti/syn, ee (anti): 86%
-
-
?
4-bromocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 29%, 81:19 anti/syn, ee (anti): 84%
-
-
?
4-chlorocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 47%, 69:31 anti/syn, ee (anti): 75%
-
-
?
4-cyanobenzaldehyde + cyclohexanone
4-[hydroxy(2-oxocyclohexyl)methyl]benzonitrile
show the reaction diagram
4-cyanoclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 60%, 72:28 anti/syn, ee (anti): 76%
-
-
?
4-methylcyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 23%, 63:37 anti/syn, ee (anti): 96%
-
-
?
4-nitrocyclohexan-1-one + acetaldehyde
?
show the reaction diagram
-
yield: 12%, ee (anti): 14%
-
-
?
4-nitrocyclohexan-1-one + cycloheptanone
?
show the reaction diagram
-
yield: 19%, 47:53 anti/syn, ee (anti): 26%
-
-
?
4-nitrocyclohexan-1-one + cyclohexanone
?
show the reaction diagram
-
yield: 69%, 63:37 anti/syn, ee (anti): 78%
-
-
?
4-nitrocyclohexan-1-one + cyclopentanone
?
show the reaction diagram
-
yield: 73%, 56:44 anti/syn, ee (anti): 52%
-
-
?
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
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-
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
albumin
-
inhibits hydrolysis of synthetic substrates
-
Cu2+
-
-
Cystatin
-
-
-
Hg2+
-
-
iodoacetate
-
-
PCMB
-
-
Succinyl-albumin
-
inhibits hydrolysis of synthetic substrates
-
Zn2+
-
-
additional information
-
crystals from potato tubers are predominantly composed of a proteinase inhibitor with a MW of 80000 Da, that inhibits chymopapain
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,3-Dimercaptopropanol
glutathione
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]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5 - 13
benzoyl-Arg-4-nitroanilide
0.0075 - 0.01
benzyloxycarbonyl-Lys phenyl ester
0.0025
benzyloxycarbonyl-Phe-Arg-4-methylcoumarin 7-amide
-
-
0.0502 - 0.0506
methyl red-Abu-Ala-Pro-Val-Lys-Lys(N5-(5-carboxyfluorescein))-NH2
0.023 - 0.026
methyl red-Abu-Ser-Ala-Pro-Val-Lys-Ala-Lys(N6-(5-carboxyfluorescein))-NH2
0.000131
Succinyl-albumin
-
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.003 - 0.05
benzoyl-Arg-4-nitroanilide
14 - 15.5
benzyloxycarbonyl-Lys phenyl ester
2.9
benzyloxycarbonyl-Phe-Arg-4-methylcoumarin 7-amide
Carica papaya
-
-
246 - 590
methyl red-Abu-Ala-Pro-Val-Lys-Lys(N5-(5-carboxyfluorescein))-NH2
133 - 360
methyl red-Abu-Ser-Ala-Pro-Val-Lys-Ala-Lys(N6-(5-carboxyfluorescein))-NH2
4
Succinyl-albumin
Carica papaya
-
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.154
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-
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.91
-
assay at. Chymopapain shows the best activity and enantioselectivity in the presence of phosphate buffer (pH 4.91, buffer/MeCN:0.12, v/v), which give the product in yield of 45% with 72:28 (anti/syn ratio) and 78% ee (for anti isomer)
5.5 - 8
-
benzoyl-L-argininamide
6.2
-
assay at
7 - 9
-
casein digestion
7
-
hemoglobin digestion
7.4
-
assay at
additional information
-
pH value of the reaction medium plays a significant role in maintaining the stability and catalytic activity of the enzyme
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 9.8
-
about 65% of maximal activity at pH 6.0 and at pH 9.8
6.2 - 7.4
-
10 min, 37°C
additional information
-
chymopapain has a broad working pH range
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
assay at
30
-
best diastereoselectivity and enantioselectivity is reached at 30°C, which give a yield of 27% with 76:24 (anti/syn ratio) and 78% ee (for anti isomer)
37
-
assay at; pH 6.2 or pH 7.4, 10 min
40
-
highest activity for aldol reaction
additional information
-
temperature is an important factor affecting the enzyme stability, selectivity and reaction rate
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
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
PDB
SCOP
CATH
ORGANISM
UNIPROT
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
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
-
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
COMMENTARY hide
LITERATURE
?
-
x * 24700, chymopapain S, SDS-PAGE
monomer
-
x * 24000, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
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
LITERATURE
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
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
667431
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
678400
2 - 3
-
enzyme undergoes conformational transition that instantaneously converts the native form into a molten globule state and is completely irreversible
667431
2
-
stable
81563
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
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
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
LITERATURE
mercury prevents irreversible oxidation
-
81571
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, freeze-dryed, stable for at least 1 month
-
0°C, pH 2.0, stable
-
4°C, 15% loss of activity after 2 weeks
-
4°C, stable in solution for at least 1 month
-
frozen, 50% loss of activity after 24 h
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; enzyme is semi-purified with from latex crude extracts by ion exchange chromatography on a CM Sephadex column C-50 and gel filtration
-
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
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
down regulated protein in papaya sticky diseased latex; down regulated protein in papaya sticky diseased latex; down regulated protein in papaya sticky diseased latex; down regulated protein in papaya sticky diseased latex
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
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