Information on EC 3.4.22.2 - papain

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.4.22.2
-
RECOMMENDED NAME
GeneOntology No.
papain
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of proteins with broad specificity for peptide bonds, but preference for an amino acid bearing a large hydrophobic side chain at the P2 position. Does not accept Val in P1'
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
acyl transfer
-
-
hydrolysis
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-
hydrolysis of peptide bond
polymerization
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CAS REGISTRY NUMBER
COMMENTARY hide
9001-73-4
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
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papain displays a strong anti-angiogenic effect in VEGF activated HUVEC human umbilical vein endothelial cells in vitro
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(RS)-mandelic hydrazide + benzyloxycarbonyl-Ala
N1-(benzyloxycarbonyl-Ala)-N2-[(R)-mandelyl]hydrazine + N1-(benzyloxycarbonyl-Ala)-N2-[(S)-mandelyl]hydrazine
show the reaction diagram
-
-
mixture of diastereoisomers containing 73% N1-(benzyloxycarbonyl-Ala)-N2-[(R)-mandelyl]hydrazine
?
(RS)-mandelic hydrazide + benzyloxycarbonyl-Gly
N1-(benzyloxycarbonyl-Gly)-N2-[(R)-mandelyl]hydrazine + N1-(benzyloxycarbonyl-Gly)-N2-[(S)-mandelyl]hydrazine
show the reaction diagram
-
-
-
?
(RS)-mandelic hydrazide + hippuric acid
?
show the reaction diagram
-
-
-
-
?
(RS)-mandelic hydrazide + N(tert-amyloxycarbonyl)-Gly
(+)-N1-(tert-amyloxycarbonyl-Gly)-NH2-[(R)-mandelyl]hydrazine + (+)-N1-(tert-anhydroxycarbonyl-Gly)-N2-[(S)-mandelyl] hydrazine
show the reaction diagram
-
-
-
?
(RS)-mandelic hydrazide + N-(tert-butyloxycarbonyl)-Gly
(+)-N1-(tert-butyloxycarbonyl-Gly)-N2[(R)-mandelyl]hydrazine + (+)-(N1)-(tert-butyloxycarbonyl-Gly)-N2[(S)-mandelyl]hydrazine
show the reaction diagram
-
-
-
-
?
2-(amino)ethyl 2'-pyridyl disulfide + H2O
?
show the reaction diagram
-
-
-
-
?
3-(amino)trimethylene 2'-pyridyl disulfide + H2O
?
show the reaction diagram
-
-
-
-
?
4(4'-dimethylaminophenylazo)benzoyl-Lys-Phe-Gly-Gly-Ala-Ala-(5-[(2-aminoethyl)amino]naphthalene) + H2O
4(4'-dimethylaminophenylazo)benzoyl-Lys-Phe-Gly + Gly-Ala-Ala-(5-[(2-aminoethyl)amino]naphthalene)
show the reaction diagram
4-(amino)-tetramethylene 2'-pyridyl disulfide + H2O
?
show the reaction diagram
-
-
-
-
?
Ac-L-Phe-Gly 4-nitroanilide + H2O
Ac-L-Phe-Gly + 4-nitroaniline
show the reaction diagram
-
whole hydrolysis process includes two stages: acylation and deacylation. The first step is a proton transfer to form a zwitterionic form (i.e. Cys-S-/His-H+ion-pair), and the second step is the nucleophilic attack on the carboxyl carbon of the substrate accompanied with the dissociation of 4-nitroaniline. The deacylation stage includes the nucleophilic attack of a water molecule on the carboxyl carbon of the substrate and dissociation between the carboxyl carbon of the substrate and the sulfhydryl sulfur of Cys25 side chain. The acylation is rate-limiting
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-
?
acetyl-Ala-Ala-Ala-p-nitroanilide + H2O
?
show the reaction diagram
-
-
-
?
acetyl-L-Phe-Gly-4-nitroanilide + H2O
acetyl-L-Phe-Gly + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
alpha-lactalbumin + H2O
?
show the reaction diagram
-
-
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?
alpha-N-benzoyl-DL-Arg-p-nitroanilide + H2O
?
show the reaction diagram
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-
-
?
azocasein + H2O
?
show the reaction diagram
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-
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?
azocasein + H2O
azopeptide + peptides
show the reaction diagram
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-
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?
benzaldehyde + acetylacetone
3-benzylidenepentane-2,4-dione
show the reaction diagram
-
35% yield after 72 h at 25C or 55% yield after 81 h at 60C. 150 mg of papain is the optimum quantity for the Knoevenagel reaction between 2 mM of benzaldehyde and 2.4 mM of acetylacetone in 5 ml of DMSO/H2O
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-
?
benzoyl arginine ethyl ester + H2O
?
show the reaction diagram
-
pH 6.3, 20C
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-
?
benzoyl-L-Arg-2-naphthylamide + H2O
benzoyl-L-Arg + 2-naphthylamine
show the reaction diagram
-
-
-
-
?
benzoyl-thiocarbamic acid + H2O
N-benzoyl-Gly + SH2
show the reaction diagram
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?
benzoyl-thiocarbamic acid ethyl ester + H2O
N-benzoyl-Gly + ethanethiol
show the reaction diagram
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-
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?
benzoyl-thiocarbamic acid methyl ester + H2O
N-benzoyl thioglycine + methanol
show the reaction diagram
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?
benzyl-Phe-Val-Arg-4-nitroanilide + H2O
benzyl-Phe-Val-Arg + 4-nitroaniline
show the reaction diagram
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-
?
benzyloxycarbonyl-Ala methyl ester + L-Arg
benzyloxycarbonyl-Ala-Arg-OH
show the reaction diagram
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?
benzyloxycarbonyl-Ala-Arg-NH2 + Arg-NH2
benzyloxycarbonyl-Ala-Arg-Arg-NH2
show the reaction diagram
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?
benzyloxycarbonyl-Ala-OMe + 4-aminoantipyrine
benzyloxycarbonyl-Ala-4-aminoantipyrine + methanol
show the reaction diagram
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-
?
benzyloxycarbonyl-Arg-Arg 4-methylcoumarin-7-amide + H2O
?
show the reaction diagram
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-
-
-
?
benzyloxycarbonyl-citrullyl-Arg 4-methylcoumarin-7-amide + H2O
?
show the reaction diagram
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?
benzyloxycarbonyl-Gly-OMe + 4-aminoantipyrine
benzyloxycarbonyl-Gly-4-aminoantipyrine + methanol
show the reaction diagram
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?
benzyloxycarbonyl-Phe-Arg 4-methylcoumarin-7-amide + H2O
?
show the reaction diagram
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?
benzyloxycarbonyl-Phe-Arg-4-methylcoumaryl-7-amide + H2O
?
show the reaction diagram
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?
benzyloxycarbonyl-Phe-Arg-4-nitroanilide + H2O
benzyloxycarbonyl-Phe-Arg + 4-nitroaniline
show the reaction diagram
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?
benzyloxycarbonyl-Phe-Leu-4-nitroanilide + H2O
benzyloxycarbonyl-Phe-Leu + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Ser-OMe + 4-aminoantipyrine
benzyloxycarbonyl-Ser-4-aminoantipyrine + methanol
show the reaction diagram
-
-
-
?
beta-lactoglobulin + H2O
?
show the reaction diagram
-
-
-
-
?
Bovine serum albumin + H2O
?
show the reaction diagram
-
-
-
-
?
carboxybenzoyl-Phe-Arg-7-(4-methyl)coumarylamide + H2O
carboxybenzoyl-Phe-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
fluorogenic substrate
-
-
?
casein + H2O
L-tyrosine + ?
show the reaction diagram
-
-
-
-
?
CBZ-beta-Ala 4-guanidinophenyl ester + L-Phe-NH2 + H2O
CBZ-beta-Ala-L-Phe-NH2 + CBZ-beta-Ala + 4-guanidinophenol
show the reaction diagram
-
-
31.6% yield of CBZ-beta-Ala-L-Phe-NH2
-
?
CBZ-D-Ala 4-guanidinophenyl ester + L-Phe-NH2 + H2O
CBZ-D-Ala-L-Phe-NH2 + CBZ-D-Ala + 4-guanidinophenol
show the reaction diagram
-
-
11.6% yield of CBZ-D-Ala-L-Phe-NH2
-
?
CBZ-Gly 4-guanidinophenyl ester + D-Phe-NH2 + H2O
CBZ-Gly-D-Phe-NH2 + CBZ-Gly + 4-guanidinophenol
show the reaction diagram
-
-
22.9% yield for CBZ-Gly-D-Phe-NH2 and 74.3% yield for CBZ-Gly
-
?
CBZ-Gly 4-guanidinophenyl ester + H2O
CBZ-Gly + 4-guanidinophenol
show the reaction diagram
-
-
94.8% yield for CBZ-Gly
-
?
CBZ-Gly 4-guanidinophenyl ester + L-Ala 4-nitroanilide + H2O
CBZ-Gly-L-Ala 4-nitroanilide + CBZ-Gly + 4-guanidinophenol
show the reaction diagram
-
-
96% yield for CBZ-Gly-L-Ala 4-nitroanilide
-
?
CBZ-Gly 4-guanidinophenyl ester + L-Ala-NH2 + H2O
CBZ-Gly-L-Ala-NH2 + CBZ-Gly + 4-guanidinophenol
show the reaction diagram
-
-
87% yield for Gly-L-Ala-NH2 and 7.7% yield for Gly-OH
-
?
CBZ-Gly 4-guanidinophenyl ester + L-Phe tert-butyl ester + H2O
CBZ-Gly-L-Phe tert-butyl ester + CBZ-Gly + 4-guanidinophenol
show the reaction diagram
-
-
11.8% yield for CBZ-Gly-L-Phe tert-butyl ester
-
?
CBZ-Gly 4-guanidinophenyl ester + L-Phe-NH2 + H2O
CBZ-Gly-L-Phe-NH2 + CBZ-Gly + 4-guanidinophenol
show the reaction diagram
-
-
92% yield of CBZ-Gly-L-Phe-NH2
-
?
CBZ-Gly 4-guanidinophenyl ester + L-Pro 4-nitroanilide + H2O
CBZ-Gly-L-Pro 4-nitroanilide + CBZ-Gly + 4-guanidinophenol
show the reaction diagram
-
-
90.3% yield for CBZ-Gly
-
?
CBZ-Gly 4-guanidinophenyl ester + L-Ser 4-nitroanilide + H2O
CBZ-Gly-L-Ser 4-nitroanilide + CBZ-Gly + 4-guanidinophenol
show the reaction diagram
-
-
94% yield for CBZ-Gly-L-Ser 4-nitroanilide
-
?
CBZ-Gly 4-guanidinophenyl ester + L-Tyr 4-nitroanilide + H2O
CBZ-Gly-L-Tyr 4-nitroanilide + CBZ-Gly + 4-guanidinophenol
show the reaction diagram
-
-
90.6% yield for CBZ-Gly-L-Tyr 4-nitroanilide and 4.3% yield for CBZ-Gly
-
?
CBZ-Gly 4-guanidinophenyl ester + L-Tyr-NH2 + H2O
CBZ-Gly-L-Tyr-NH2 + CBZ-Gly + 4-guanidinophenol
show the reaction diagram
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-
91.3% yield for CBZ-Gly-L-Tyr-NH2 and 2.5% yield for CBZ-Gly
-
?
CBZ-L-Ala 4-guanidinophenyl ester + L-Phe-NH2 + H2O
CBZ-L-Ala-L-Phe-NH2 + CBZ-L-Ala + 4-guanidinophenol
show the reaction diagram
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-
77.5% yield of CBZ-L-Ala-L-Phe-NH2
-
?
CBZ-L-Arg 4-guanidinophenyl ester + L-Phe-NH2 + H2O
CBZ-L-Arg-L-Phe-NH2 + CBZ-L-Arg + 4-guanidinophenol
show the reaction diagram
-
-
45.9% yield of CBZ-L-Arg-L-Phe-NH2
-
?
CBZ-L-Asn 4-guanidinophenyl ester + L-Phe-NH2 + H2O
L-Asn-L-Phe-NH2 + CBZ-L-Asn + 4-guanidinophenol
show the reaction diagram
-
-
6.1% yield of CBZ-L-Asn-L-Phe-NH2
-
?
CBZ-L-Glu 4-guanidinophenyl ester + L-Phe-NH2 + H2O
CBZ-Glu-L-Phe-NH2 + CBZ-L-Glu + 4-guanidinophenol
show the reaction diagram
-
-
68.5% yield of CBZ-L-Glu-L-Phe-NH2
-
?
CBZ-L-Ile 4-guanidinophenyl ester + L-Phe-NH2 + H2O
CBZ-L-Ile-L-Phe-NH2 + CBZ-L-Ile + 4-guanidinophenol
show the reaction diagram
-
-
24.5% yield of CBZ-L-Ile-L-Phe-NH2
-
?
CBZ-L-Thr 4-guanidinophenyl ester + L-Phe-NH2 + H2O
CBZ-L-Thr-L-Phe-NH2 + CBZ-L-Thr + 4-guanidinophenol
show the reaction diagram
-
-
90.7% yield of CBZ-L-Thr-L-Phe-NH2
-
?
CH3-CH2-2-pyridyl disulfide + H2O
?
show the reaction diagram
-
-
-
-
?
CH3CO-(D-Phe)-NH-[CH2]2-2-pyridyl disulfide + H2O
?
show the reaction diagram
-
-
-
-
?
CH3CO-(D-Phe)-O-[CH2]2-2-pyridyl disulfide + H2O
?
show the reaction diagram
-
-
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?
CH3CO-(L-Phe)-NH-[CH2]2-2-pyridyl disulfide + H2O
?
show the reaction diagram
-
-
-
-
?
CH3CO-(L-Phe)-O-[CH2]2-2-pyridyl disulfide + H2O
?
show the reaction diagram
-
-
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?
CH3CO-NH-[CH2]2-2-pyridyl disulfide + H2O
?
show the reaction diagram
-
-
-
-
?
CH3CO-O-[CH2]2-2-pyridyl disulfide + H2O
?
show the reaction diagram
-
-
-
-
?
chicken IgY + H2O
?
show the reaction diagram
-
-
-
-
?
chitosan + H2O
low-molecular mass chitosan + chito-oligomeric-monomeric mixture
show the reaction diagram
CopA + H2O
?
show the reaction diagram
-
CopA is a bacterial Cu+-ATPase from Thermotoga maritima and contains 3 papain cleavage sites on the C-terminal side of the N-terminal metal binding domain
-
-
?
curbitin + H2O
?
show the reaction diagram
-
the reaction occurs in two successive steps. In the first step, limited proteolysis consisting of detachments of short terminal peptides from the alpha and beta chains are observed. The cooperative proteolysis, which occurs as a pseudo-first order reaction, started at the second step. The limited proteolysis at the first step plays a regulatory role, impacting the rate of deep degradation of cucurbitin molecules by the cooperative mechanism
-
-
?
DL-4-hydroxyphenylglycine methyl ester + H2O
?
show the reaction diagram
-
asymmetric hydrolysis
-
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?
fibroin + H2O
?
show the reaction diagram
-
-
upon papain hydrolysis of fibroin composed of highly repetitive Ala- and Gly-rich blocks even-numbered peptides are obtained. The even-numbered peptides are in the forms of di-, tetra-, hexa-, and octa-peptides with repeating units in combination of Ala-Gly, Ser-Gly, Tyr-Gly, and Val-Gly. The sequences of the tetra-peptides are in the order of Ala-Gly-X-Gly, where X is Tyr or Val
-
?
fish IgM + H2O
?
show the reaction diagram
-
-
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?
Glucagon + H2O
?
show the reaction diagram
-
-
-
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?
Hemoglobin + H2O
?
show the reaction diagram
-
alpha-chain and beta-chain
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?
hippuric acid + aniline
hippuryl anilide
show the reaction diagram
-
weak activity, 0.1% of the hydrolytic activity with N-benzoyl-L-argininamide
-
r
human IgG + H2O
fragment Fab + fragment Fc
show the reaction diagram
-
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-
?
immunoglobulin M + H2O
IgMI +
show the reaction diagram
-
release of a basic subunit-like fragment which is designated IgMI, by proteolysis of the m-chain near the carboxyl terminus
-
?
L-Arg-7-amido-4-methylcoumarin + H2O
L-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
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?
L-glutamic acid diethyl ester + L-glutamic acid diethyl ester
L-Glu-gamma-diethyl ester polymer
show the reaction diagram
-
polymerization reaction
-
-
?
L-glutamic acid diethyl ester + L-glutamic acid diethyl ester
oligo-gamma-ethyl-L-glutamate
show the reaction diagram
-
oligomerization reaction
-
-
?
L-glutamic acid diethyl ester + N-alpha-benzoyl-L-arginine ethyl ester
N-alpha-benzoyl-L-arginine + N-alpha-benzoyl-L-argininyl-Glu-diethyl ester
show the reaction diagram
-
-
-
-
?
L-glutamic acid triethyl ester + N-alpha-benzoyl-L-arginine ethyl ester
N-alpha-benzoyl-L-arginine + N-alpha-benzoyl-L-argininyl-Glu-Glu-triethyl ester
show the reaction diagram
-
L-glutamic acid triethyl ester shows higher affinity for papain than L-glutamic acid diethyl ester
-
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?
L-phenylalanine amide + H2O
L-phenylalanine + NH3
show the reaction diagram
-
-
-
-
?
L-Pro-L-Phe-L-Leu-4-nitroanilide + H2O
L-Pro-L-Phe-L-Leu + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
L-Pro-Phe-Leu-4-nitroanilide + H2O
L-Pro-Phe-Leu + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
lambda repressor + H2O
?
show the reaction diagram
-
no cleavage of the operator-bound repressor dimer
-
?
lipid transfer protein + H2O
?
show the reaction diagram
-
-
-
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?
low molecular weight heparin + H2O
?
show the reaction diagram
-
-
-
-
?
methyl red-Abu-Ala-Pro-Val-Lys-Lys(N5-(5-carboxyfluorescein))-NH2 + H2O
?
show the reaction diagram
-
pH 6.2 or pH 7.4, 10 min, 37C
-
-
?
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
-
-
?
methyl red-Abu-Ser-Ala-Pro-Val-Lys-Ala-Lys(N5-(5-carboxyfluorescein))-NH2 + H2O
?
show the reaction diagram
-
pH 6.2 or pH 7.4, 10 min, 37C
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-
?
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(beta-phenylpropionyl)Gly methyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
N,N-diBoc-dityrosine-(isoniazid)2 + H2O
?
show the reaction diagram
-
-
-
-
?
N-acetyl-L-Trp p-nitrophenyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
N-acetyl-L-tyrosinamide + H2O
N-acetyl-L-Tyr + NH3
show the reaction diagram
-
-
-
-
?
N-alpha-benzoyl-DL-Arg-4-nitroanilide + H2O
N-alpha-benzoyl-DL-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
N-alpha-benzoyl-L-Arg-4-nitroanilide + H2O
N-alpha-benzoyl-L-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
N-alpha-benzyloxycarbonyl-L-lysine 4-nitrophenyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzoyl-Arg-p-nitroanilide + H2O
Nalpha-benzoyl-Arg + p-nitroanilide
show the reaction diagram
-
-
-
?
N-benzoyl-DL-arginine-2-naphthylamide + H2O
N-benzoyl-DL-arginine + 2-naphthylamine
show the reaction diagram
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-
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?
N-benzoyl-Gly ethyl ester + H2O
N-benzoyl-Gly + ethanol
show the reaction diagram
-
-
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?
N-benzoyl-Gly methyl ester + H2O
?
show the reaction diagram
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?
N-benzoyl-Gly methyl ester + H2O
N-benzoyl-Gly + methanol
show the reaction diagram
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-
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?
N-benzoylglycinamide + H2O
Nalpha-benzoyl-Gly + NH3
show the reaction diagram
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?
N-benzyloxycarbonyl-Ala methyl ester + H2O
?
show the reaction diagram
-
-
-
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?
N-benzyloxycarbonyl-Gly 2-nitrophenyl ester + H2O
N-benzyloxycarbonyl-Gly + 2-nitrophenol
show the reaction diagram
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-
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?
N-benzyloxycarbonyl-Gly 3-nitrophenyl ester + H2O
N-benzyloxycarbonyl-Gly + 3-nitrophenol
show the reaction diagram
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-
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?
N-Benzyloxycarbonyl-Gly 4-nitrophenyl ester + H2O
N-Benzyloxycarbonyl-Gly + 4-nitrophenol
show the reaction diagram
-
-
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?
N-benzyloxycarbonyl-Gly ethyl ester + H2O
?
show the reaction diagram
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-
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?
N-benzyloxycarbonyl-Gly phenyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-Gly-Gly + H2O
?
show the reaction diagram
-
-
-
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?
N-benzyloxycarbonyl-Gly-p-nitroanilide + H2O
?
show the reaction diagram
-
-
-
?
N-benzyloxycarbonyl-L-Glu diamide + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-L-glycine + H2O
?
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-L-histidinamide + H2O
Nalpha-benzoyl-L-His + NH3
show the reaction diagram
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-
-
-
?
N-benzyloxycarbonyl-L-leucinamide + H2O
Nalpha-benzoyl-L-Leu + NH3
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-L-Lys + H2O
?
show the reaction diagram
-
-
-
-
?
n-propyl 2-pyridyl disulfide + H2O
?
show the reaction diagram
-
-
-
-
?
Nalpha-benzoyl-Arg-p-nitroanilide + H2O
Nalpha-benzoyl-Arg + p-nitroaniline
show the reaction diagram
-
-
-
?
Nalpha-benzoyl-DL-Arg-4-nitroanilide + H2O
Nalpha-benzoyl-DL-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
?
Nalpha-benzoyl-DL-arginine-4-nitroanilide + H2O
Nalpha-benzoyl-DL-arginine + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
Nalpha-benzoyl-L-Arg ethyl ester
?
show the reaction diagram
-
-
-
-
-
Nalpha-Benzoyl-L-Arg ethyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
Nalpha-benzoyl-L-argininamide + H2O
Nalpha-benzoyl-L-Arg + NH3
show the reaction diagram
-
-
-
-
?
Nalpha-benzoyl-L-arginine ethyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
Nalpha-benzoyl-L-citrulline methyl ester + H2O
?
show the reaction diagram
-
-
-
-
?
Nalpha-benzoyl-L-lysinamide + H2O
Nalpha-benzoyl-L-Lys + NH3
show the reaction diagram
-
-
-
-
?
ovalbumin + H2O
?
show the reaction diagram
-
-
-
-
?
oxidized beta-chain of insulin + H2O
?
show the reaction diagram
-
-
-
-
?
Phe-Arg-4-methylcoumaryl-7-amide + H2O
?
show the reaction diagram
-
-
-
?
phthalyl-Phe-Leu-p-nitroanilide + H2O
phthalyl-Phe-Leu + 4-nitroaniline
show the reaction diagram
-
-
-
?
rabbit IgG + H2O
?
show the reaction diagram
-
-
-
-
?
sarcoendoplasmic reticulum Ca2+-ATPase 1 + H2O
?
show the reaction diagram
-
-
-
-
?
sheep IgG + H2O
?
show the reaction diagram
-
-
-
-
?
succinyl-Phe-Leu-4-methylcoumaryl-7-amide + H2O
?
show the reaction diagram
-
-
-
?
succinyl-Phe-Leu-4-nitroanilide + H2O
succinyl-Phe-Leu + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
succinyl-Phe-Leu-p-nitroanilide + H2O
?
show the reaction diagram
-
-
-
?
succinyl-Phe-Leu-p-nitrophenol + H2O
?
show the reaction diagram
-
-
-
?
tarocystatin + H2O
?
show the reaction diagram
-
the C-terminal cystatin-like extension of tarocystatin is easily digested by papain
-
-
?
ubiquitin-7-amido-4-trifluoromethylcoumarin + H2O
?
show the reaction diagram
-
-
-
-
?
Z-Phe-Arg-4-nitroanilide + H2O
Z-Phe-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
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
casein + H2O
?
show the reaction diagram
-
-
-
-
?
chitosan + H2O
low-molecular mass chitosan + chito-oligomeric-monomeric mixture
show the reaction diagram
-
depolymerization, the enzyme inhibits the growth of bacteria such as Bacillus cereus strain F4810, Bacillus licheniformis, and Escherichia coli strain D21, mechanism of bactericidal action of the chito-oligomeric-monomeric mixture, overview
-
-
?
tarocystatin + H2O
?
show the reaction diagram
-
the C-terminal cystatin-like extension of tarocystatin is easily digested by papain
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
1 mM, enzymatic activity increases to a maximum of 18%
Mg2+
-
1 mM, enzymatic activity increases to a maximum of 24%
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2-[(S,R)-2-oxo-4-phenylazetidin-1-yl]acetyl)-L-phenylalanine methyl ester
-
weak inhibitor, irreversible
(2-[(S,R)-2-oxo-4-phenylazetidin-1-yl]acetyl)-L-Val benzyl ester
-
weak inhibitor, irreversible
(eta5-C5H5)Fe(CO)3 eta1-N-succinimidato
-
metallocarbonyl complex, reversible inhibitor
-
(eta5-C5H5)Mo(CO)3 eta1-N-succinimidato
-
metallocarbonyl complex, reversible inhibitor
-
(eta5-C5H5)W(CO)3 eta1-N-succinimidato
-
metallocarbonyl complex, reversible inhibitor
-
(S)-1-[(S)-N-(tert-butyloxycarbonyl)alanyl]-4-oxoazetidine-2-carboxylic acid
-
weak, irreversible inactivation
(S,R)-1-[(S)-N-(tert-butyloxycarbonyl)alanyl]-4-phenylazetidin-2-one
-
weak inhibitor, irreversible
1,1-dicyano-2-(4,5-dimethoxy-2-nitrophenyl)-ethene
-
irreversible, photosensitive inhibitor
1,3-Dibromoacetone
-
-
1-(4,5-dimethoxy-2-nitrophenyl)-2-nitroethene
-
irreversible, photosensitive inhibitor
2-Pentanone
-
noncompetitive
acetyl-Phe-Gly-S-nitrosopenicillamine
-
inactivation of papain by S-nitrosothiols is due to a direct attack of the highly reactive thiolate Cys25 in the enzyme active site on the sulfur of S-nitrosothiols to form a mixed disulfide between the inactivator and papain
antipain
-
-
ascorbic acid
-
strong
Ba2+
-
residual activity in the presence of 20 mM: 13% free papain, 38% immobilized papain
barley cystatin protease inhibitor
-
inhibited by various cystatin vatiants, HvCPI-1, HvCPI-2, HvCPI-3, HvCPI-4, HvCPI-5, HvCPI-6
-
benzoylarginine
-
competitive
benzoylglycine
-
competitive
benzyl-(S)-1-[(S)-N-(tert-butyloxycarbonyl)alanyl]-4-oxoazetidine-2-carboxylate
-
weak, irreversible inactivation
benzyl-(S)-2-(benzyloxycarbonyl)azetidin-1-acetate
-
weak, irreversible inactivation
benzyloxycarbonyl-Arg-Leu-Val-Gly-CHN2
-
-
benzyloxycarbonyl-Phe-Ala-glyoxal
-
competitive, 13C-NMR study of the inhibition
Bromoacetic acid
-
-
Ca2+
-
residual activity in the presence of 20 mM: 10% free papain, 48% immobilized papain
carbobenzyloxy-leucinyl-leucinyl-leucinal
-
-
chagasin
Chloroacetate
-
-
CNWAAGYNCGGGS-NH2
-
synthetic cyclic peptide, cyclization through intramolecular disulfide bonding
CNWTLGGYKCGGGS-NH2
-
synthetic cyclic peptide, cyclization through intramolecular disulfide bonding
CpPRI
-
pathogenesis-related class 10 protein with noncompetitive papain inhibitory activity, purified from Crotalaria pallida roots. CpPRI is made up of a single polypeptide chain with a Mr of 15 kDa
-
Cu2+
-
residual activity in the presence of 20 mM: 0% free papain, 20% immobilized papain
CWEWGGWHCGGSS-OH
-
synthetic cyclic peptide, cyclization through intramolecular disulfide bonding
CWSMMGFQCGGGS-NH2
-
weak inhibition, synthetic cyclic peptide, cyclization through intramolecular disulfide bonding
Cystatin
-
-
-
cystatin B
-
-
-
cystatin SN
-
-
-
cystatin SN variant G12A/G13A
-
-
-
cystatin SN variant P106G/W107G
-
-
-
cystatin SN variant Y56G/T57G/V58G
-
-
-
diethyldisulfide-beta,beta'-dicarboxylic acid
-
-
-
dimethyl sulfoxide
-
the number of active sites of papain decreases with increasing concentration of dimethyl sulfoxide whereas the incubation time, in a buffer containing 3% dimethyl sulfoxide does not affect the number of active sites. A rapid decrease of the initial reaction rate, by up to 30%, is observed between 1 and 2% dimethyl sulfoxide
dimethylformamide
-
number of papain active sites decreases with increase of inhibitor concentration
E-64
-
-
endopin 2
-
highly effective inhibition, cross-class inhibition of papain and elastase. Localization of endopin 2 to regulated secretory vesicles of neuroendocrine chromaffin cells
-
ethanol
-
activity decreases with increasing ethanol content, up to 15% ethanol papain from papaya latex is less sensitive to ethanol
ethyl-(RS)-2-(2-oxo-4-phenylazetidin-1-yl)acetate
-
weak inhibitor, irreversible
glucose-2S-nitroso-N-acetyl-penicillamine
-
inactivation of papain by S-nitrosothiols is due to a direct attack of the highly reactive thiolate Cys25 in the enzyme active site on the sulfur of S-nitrosothiols to form a mixed disulfide between the inactivator and papain
GNWTLGGYKGG
-
weak inhibition, synthetic cyclic peptide, cyclization head-to-tail
HgCl2
-
-
inhibitor of cysteine protease
-
-
-
iodoacetamide
-
-
iodoacetic acid
-
-
kininogen2
-
from bovine, rat or human
-
Kunitz type trypsin inhibitor
-
i.e. PTPKI, SwissProt: P32722 (alpha chain), P32773 (beta-chain), a small Kunitz trypsin inhibitor from Prosopis juliflora, 0.025 mg/ml, 98.3% inhibition, overlapping binding sites for trypsin and papain
-
L-Ala-L-Phe-aminoacetonitrile
-
-
Leupeptin
-
-
methanol
-
number of papain active sites decreases with increase of inhibitor concentration
methyl methanethiosulfonate
-
-
Mg2+
-
residual activity in the presence of 20 mM: 20% free papain, 57% immobilized papain
Mn2+
-
residual activity in the presence of 20 mM: 0% free papain, 18% immobilized papain
N-benzyloxycarbonyl-L-Phe-L-Ala chloromethyl ketone
-
-
N-methylcyrhetrene-pyrrole-2,5-dione
-
-
N-methylferrocene-pyrrole-2,5-dione
-
-
N-methylruthenocene-pyrrole-2,5-dione
-
-
N-[N-(L-3-trans-carboxyoxirane-2-carbonyl)-L-leucyl]-isoamylamide
-
-
-
Ni2+
-
residual activity in the presence of 20 mM: 0% free papain, 16% immobilized papain
oryzacastatin
-
and fragments. The NH2-terminal 21 rsidues including Gly5 and the COOH-terminal 11 residues of the inhibitor are not essential for inhibition
-
p-CH3-C6H4(SO2)-NH-CH2-CO-CH2Cl
-
-
papain inhibitors
-
A1, A2, A3, B2 and C from seeds of Vigna unguiculata subsp. cylindrica
-
PdKl-3.1
-
peptide inhibitor purified from seed of Pithecellobium dumosum tree, stable over a wide range of pH and temperature. Inhibitory to trypsin, moderately inhibitory to papain
-
PdKl-3.2
-
peptide inhibitor purified from seed of Pithecellobium dumosum tree, stable over a wide range of pH and temperature. Inhibitory to trypsin, moderately inhibitory to papain
-
S-nitroso-N-acetyl-DL-penicillamine
-
inactivation of papain by S-nitrosothiols is due to a direct attack of the highly reactive thiolate Cys25 in the enzyme active site on the sulfur of S-nitrosothiols to form a mixed disulfide between the inactivator and papain
S-nitrosocaptopril
-
inactivation of papain by S-nitrosothiols is due to a direct attack of the highly reactive thiolate Cys25 in the enzyme active site on the sulfur of S-nitrosothiols to form a mixed disulfide between the inactivator and papain
S-nitrosoglutathione
-
inactivation of papain by S-nitrosothiols is due to a direct attack of the highly reactive thiolate Cys25 in the enzyme active site on the sulfur of S-nitrosothiols to form a mixed disulfide between the inactivator and papain
staccopin P1
-
-
-
staccopin P2
-
-
-
strepin P-1
-
-
-
succinyl-Ala-Val-Val-Ala-Ala-p-nitroanilide
-
-
succinyl-Gln-Val-Val-Ala-Ala methyl ester
-
-
succinyl-Gln-Val-Val-Ala-Ala-p-nitroanilide
-
-
succinyl-Gln-Val-Val-Ala-p-nitroanilide
-
-
succinyl-Gln-Val-Val-p-nitroanilide
-
weak inhibition
tarocystatin
-
the N-terminal cystatin domain (residues 1-98) of tarocystatin has inhibitory ability against papain
-
Urea
-
the reversible unfolding/refolding process of the PLpro in the presence of urea is investigated. The zinc-binding domain of the enzyme starts to unfold at urea concentration below 0.35 M and reaches a first plateau at 1-3.5 M. At this stage, the palm and thumb core domains remain well folded but totally inactive
Zn2+
-
residual activity in the presence of 20 mM: 0% free papain, 16% immobilized papain
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptopropionic acid
-
required, complete activation at 0.05 N when the enzyme concentration is 0.49 mM
Cys
-
activates
cysteine
-
the requirement for reducing conditions during the activation process is investigated with 20 mM DTT or 20 mM cysteine, propapain does not convert to papain even after prolonged incubation with DTT, whereas in presence of 20 mM cysteine, the activation occurs between 30 and 40 min at 50C
EDTA
-
required for maximal activity
glutathione
-
-
Reducing agents
-
e.g. 2,3-dimercaptopropanol, activates
-
Sodium borohydride
-
activates
thiol compounds
-
activate
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00385 - 1.765
4(4'-dimethylaminophenylazo)benzoyl-Lys-Phe-Gly-Gly-Ala-Ala-(5-[(2-aminoethyl)amino]naphthalene)
0.048
alpha-benzoyl-L-argininamide
-
with Cys as activator
0.77
benzoyl-thiocarbamic acid
-
-
16
benzoyl-thiocarbamic acid ethyl ester
-
-
0.3
benzoyl-thiocarbamic acid methyl ester
-
-
0.221 - 0.616
benzyl-Phe-Val-Arg-4-nitroanilide
0.94 - 5.6
benzyloxycarbonyl-Arg-Arg 4-methylcoumarin-7-amide
1.296 - 2.03
benzyloxycarbonyl-citrullyl-Arg 4-methylcoumarin-7-amide
1.8
benzyloxycarbonyl-L-citrullyl-L-Arg 4-methylcoumarin-7-amide
-
mutant enzyme D158N
0.4 - 0.42
benzyloxycarbonyl-L-Phe-L-Arg 4-methylcoumarin-7-amide
0.089 - 1.48
benzyloxycarbonyl-Phe-Arg 4-methylcoumarin-7-amide
0.06
carboxybenzoyl-Phe-Arg-7-(4-methyl)coumarylamide
-
100 mM sodium phosphate buffer, pH 6.0, containing 1 mM dithiothreitol and 1 mM EDTA
0.0000082 - 812.1
casein
0.119 - 0.2
methyl red-Abu-Ala-Pro-Val-Lys-Lys(N5-(5-carboxyfluorescein))-NH2
0.027 - 0.1015
methyl red-Abu-Ser-Ala-Pro-Val-Lys-Ala-Lys(N6-(5-carboxyfluorescein))-NH2
0.00691
N,N-diBoc-dityrosine-(isoniazid)2
-
pH 6.2, 25C
-
6.63
N-(beta-phenylpropionyl)Gly methyl ester
-
-
0.11
N-(beta-phenylpropionyl)Gly methyl thiono ester
-
-
-
0.00212
N-alpha-benzyloxycarbonyl-L-lysine 4-nitrophenyl ester
-
pH 6.5, 27C, Vmax: 2.14 miroM/sec
-
21.8
N-benzoyl-Gly methyl ester
-
-
0.301
N-benzoyl-Gly methyl thiono ester
-
-
-
21
N-benzoylglycinamide
-
38C
0.008
N-benzyloxycarbonyl-Gly p-nitrophenyl ester
-
-
320
N-benzyloxycarbonyl-Gly-Gly
-
-
18.54
Nalpha-benzoyl-Gly methyl ester
-
-
0.0519 - 23
Nalpha-benzoyl-L-Arg ethyl ester
32 - 40
Nalpha-benzoyl-L-argininamide
10
Nalpha-benzoyl-L-citrulline methyl ester
-
-
20
Nalpha-benzyloxycarbonyl-L-histidinamide
-
-
0.73 - 1.84
ubiquitin-7-amido-4-trifluoromethylcoumarin
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.03 - 6.86
4(4'-dimethylaminophenylazo)benzoyl-Lys-Phe-Gly-Gly-Ala-Ala-(5-[(2-aminoethyl)amino]naphthalene)
3340
benzoyl-thiocarbamic acid
Carica papaya
-
-
3.4
benzoyl-thiocarbamic acid ethyl ester
Carica papaya
-
-
0.064
benzoyl-thiocarbamic acid methyl ester
Carica papaya
-
-
180 - 309
benzyl-Phe-Val-Arg-4-nitroanilide
0.79 - 9.3
benzyloxycarbonyl-Arg-Arg 4-methylcoumarin-7-amide
0.42 - 8.14
benzyloxycarbonyl-citrullyl-Arg 4-methylcoumarin-7-amide
51 - 52
benzyloxycarbonyl-L-Phe-L-Arg 4-methylcoumarin-7-amide
2.78 - 41.6
benzyloxycarbonyl-Phe-Arg 4-methylcoumarin-7-amide
0.00000028 - 1.79
casein
475 - 1773
methyl red-Abu-Ala-Pro-Val-Lys-Lys(N5-(5-carboxyfluorescein))-NH2
243 - 1503
methyl red-Abu-Ser-Ala-Pro-Val-Lys-Ala-Lys(N6-(5-carboxyfluorescein))-NH2
35.6
N-alpha-benzyloxycarbonyl-L-lysine 4-nitrophenyl ester
Carica papaya
-
pH 6.5, 27C
-
3.48
N-benzoyl-Gly methyl ester
Carica papaya
-
-
16.4 - 28.5
Nalpha-benzoyl-L-Arg ethyl ester
0.79 - 0.96
ubiquitin-7-amido-4-trifluoromethylcoumarin
additional information
additional information
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00035 - 0.0076
casein
16700
N-alpha-benzyloxycarbonyl-L-lysine 4-nitrophenyl ester
Carica papaya
-
pH 6.5, 27C
206877
0.52 - 1.08
ubiquitin-7-amido-4-trifluoromethylcoumarin
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.096
acetyl-Phe-Gly-S-nitrosopenicillamine
-
pH 6.2, 25C
0.0000033
benzyloxycarbonyl-Phe-Ala-glyoxal
-
pH 7.03, 25C
0.000000036
chagasin
-
inactivation of papain is very fast, rate constant for inactivation is 1500 mM/s
-
0.0063
CNWAAGYNCGGGS-NH2
-
pH 6.8, 37C
0.0286
CNWTLGGYKCGGGS-NH2
-
pH 6.8, 37C
0.0000018
CpPRI
-
pH 7.5, 37C
-
0.0071
CWEWGGWHCGGSS-OH
-
pH 6.8, 37C
0.00000001
cystatin SN, cystatin SN variant G12A/G13A, cystatin SN variant P106G/W107G
-
below
-
0.000057
cystatin variant Y56G/T57G/V58G
-
-
-
0.618
D-glucose-2S-nitroso-N-acetyl-penicillamine
-
pH 6.2, 25C
0.0000019
papain prosegment
recombinant prosegment, pH 7.0, 20C, inhibition kinetics
-
0.356
S-nitroso-N-acetyl-DL-penicillamine
-
pH 6.2, 25C
0.449
S-nitrosocaptopril
-
pH 6.2, 25C
0.305
S-nitrosoglutathione
-
pH 6.2, 25C
additional information
additional information
-
inhibition constants for various barley cystatin protease inhibitor variants
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.7
(eta5-C5H5)Fe(CO)3 eta1-N-succinimidato
Carica papaya
-
pH 6.5, temperature not specified in the publication
-
0.28
(eta5-C5H5)Mo(CO)3 eta1-N-succinimidato
Carica papaya
-
pH 6.5, temperature not specified in the publication
-
0.48
(eta5-C5H5)W(CO)3 eta1-N-succinimidato
Carica papaya
-
pH 6.5, temperature not specified in the publication
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.059
-
activity of immobilized papain dehydrated by n-propanol in low-water tert-butanol media
1.65
-
purified enzyme at pH 10.5 and 60C
1.78
-
purified enzyme, substrate chitosan
121
-
mutant K174R/V32S, pH 6.5, 37C
125.9
-
recombinant papain, in 100 mM sodium acetate and 2 mM EDTA at pH 6.5
128
-
wild-type, pH 6.5, 37C
130
-
mutant K174R/V32S/G36S, pH 6.5, 37C
221.7
-
commercial papain, in 100 mM sodium acetate and 2 mM EDTA at pH 6.5
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.2
-
assay at
3.5
-
depolymerization of chitosan
6 - 6.5
-
-
6.2
-
assay at
6.3
-
20C
6.4
-
assay at
6.6
assay at
6.8
-
assay at
6.9
-
optimal pH free papain
7.1
-
The effect of initial pH and temperature on adsorption capacity for papain on the chitosan-coated nylon composite membranes in Tris-HCl buffer is tested, an optimum point of maximum adsorption capacity obtained as 26.61 mg/g at 39C, pH 7.05 by fixing 8 mg/ml initial papain concentration. Adsorption capacity of papain increases from 26.61 to 27.85 mg/g when the papain concentration increases from 8 to 11 mg/ml.
7.2
-
hydrolysis of alpha-N-benzoyl-L-Arg ethyl ester, soluble enzyme
8.2
-
hydrolysis of alpha-N-benzoyl-L-Arg ethyl ester, immobilized enzyme
9.5
-
hydrolysis of benzyloxycarbonyl-Ala methyl ester
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.8 - 4.5
-
conditions for in vitro activation of the refolded propapain to mature papain are determined, the highest rates of activation are observed between pH 3.8 and 4.5, while the rates are greatly reduce at pH higher than 6
4 - 8
-
-
4.1 - 8.5
-
-
5 - 10
-
pH profile of soluble and immobilized enzyme
5 - 8
-
pH 5.0: about 70% of maximal activity, pH 8.0: about 85% of maximal activity
6.2 - 7.4
-
10 min, 37C
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
27
-
assay at
50
-
wild-type
70
-
optimal pH immobilized papain
85
-
immobilized enzyme
additional information
-
temperature-dependences of the second-order rate constants of the reaction involving the catalytic site thiol, overview, stopped-flow method
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 30
-
-
20 - 90
-
temperature profile of soluble and immobilized enzyme
25 - 70
-
25C: about 65% of maximal activity, 70C: about 60% of maximal acticity
50 - 100
-
50C: about 35% of maximal activity, 100C: about 95% of maximal activity, immobilized enzyme
60 - 80
-
the activity of immobilized unmodified papain and immobilized benzenetricarboxylic papain are increased stably over a period of 60 to 80C without significant change
80
-
no hydrolysis at all at temperatures higher than 80C
additional information
-
-
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.4
recombinnat prosegment, isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
maximal concentration is reached early in the development of the fruit
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
20700
-
enzyme from commercial chymopapain, equilibrium measurement
23000
-
SDS-PAGE
23700
-
equilibrium sedimentation
23800
-
enzyme from spray-dried latex, equilibrium measurement
42000
-
propapain containing respective fusion-tag sequence, determined by SDS-PAGE
additional information
-
during processing of propapain to papain, 2 intermediates are observed in the range of 30-38 kDa at 20 min incubation at 50C indicating that processing of propapain to papain occurs in a stepwise manner
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
no modification
-
nonglycosylated enzyme
proteolytic modification
the proenzyme is activated by cleavage of the prosegment
side-chain modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a complex of chagasin, a protein inhibitor from Trypanosoma cruzi, and papain is crystallized by using the hanging-drop vapor diffusion method at 18C, the high-resolution crystal structure shows an inhibitory wedge comprising three loops, which forms a number of contacts responsible for the high-affinity binding, the present chagasin-papain complex provides a reliable model of chagasin-cruzipain interactions
-
complex with benzyloxycarbonyl-Arg-Leu-Val-Gly-CHN2
-
complex with E-64; enzyme structure of the complex of papain with N-[N-(L-3-trans-carboxyoxirane-2-carbonyl)-L-leucyl]-isoamylamide
-
crystal structure of the complex of papain with recombinant human stefin B
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crystal structure of the papain-succinyl-Gln-Val-Val-Ala-Ala-p-nitroanilide complex at 1.7 -A resolution
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enzyme structure of the complex of papain with N-[N-(L-3-trans-carboxyoxirane-2-carbonyl)-L-leucyl]-isoamylamide
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enzyme structure of the complex of papain with N-[N-(L-3-trans-carboxyoxirane-2-carbonyl)-L-leucyl]-isoamylamide; structure of papain complexed with benzyloxycarbonyl-L-Phe-L-Ala-L-Ala chloromethylketone
-
hanging drop vapour diffusion method with 50% ethanol, 0.01 M sodium acetate, crystallization of a papain-inhibitor of protease complex is not possible
-
papain complexed with cathepsin B-specific covalent-type inhibitor
-
papain-inhibitor complex with benzyloxycarbonyl-Leu-Leu-leucinal or benzyloxycarbonyl-L-Leu-L-Leu methoxymethyl ketone
-
papain-leupeptin complex
-
tarocystatin-papain complex and C-terminal cystatin-like extension-papain complex, hanging drop vapor diffusion method, the tarocystatin-papain complex is crystallized from a drop containing 15% PEG monomethyl ether 2000, 0.05 M sodium acetate trihydrate, pH 4.6, 0.1 M ammonium sulfate against a reservoir of 30% PEG monomethyl ether 2000, 0.1 M sodium acetate trihydrate, pH 4.6, and 0.2 M ammonium sulfate. The complex of CtE-papain is crystallized from a drop containing 0.05 M HEPES, pH 7.5, and 35% (v/v) 2-methyl-2,4-pentanediol against a reservoir of 0.1 M HEPES, pH 7.5, and 70% (v/v) 2-methyl-2,4-pentanediol
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1 - 2
-
half life after 60 storage: below 5% free papain, 40% immobilized papain
733073
2 - 7
-
25C, stable at pH 7.0, acid unfolding at pH 2.0
667306
3
-
half life after 60 storage: below 5% free papain, 40% immobilized papain
733073
4 - 10
-
stable
95685
4
-
rapid and irreversible inactivation, at elevated temperatures
95663
6 - 11
-
the native enzyme rapidly decreases to the relative activity of 70%
695700
7 - 10
-
the native papain displaying over 80% of its activity; the papain modified by by 1,2,4-benzenetricarboxylic anhydride and pyromellitic anhydride displaying over 80% of its activity
695700
7
-
native papain loses its activity gradually above pH 7.0
681479
10
-
residual activity: 20% free papain, 50% immobilized papain
733073
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
residual activity after 30 storage: 0% free papain, 80% immobilized papain
25
-
pH 7.0, 1 h, 49.8% loss of activity
30
-
residual activity: 70% free papain, 40% immobilized papain
35.3
-
pH 2.0, first transition midpoint in absence of urea
44.2
-
pH 2.0, first transition midpoint in presence of 0.8 M urea
50
-
oxidized, inactive enzyme, in absence of activator, 67% of the initial activity remains after 28 days
53.3
-
pH 2.0, second transition midpoint in absence of urea
56
-
at pH 2.0, partially reduced papain, single cooperative transition with a midpoint at 55.61C
66.9
-
pH 2.0, second transition midpoint in presence of 0.8 M urea
85
-
10 min, in 2 mM EDTA solution with 0.08 M Cys, increase in activity
95
-
10 min, in 2 mM EDTA solution with 0.08 M Cys, about 80% loss of activity
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
chemical modification using citraconic acid, phthalic acid, maleic acid, and succinic acid leads to an increased thermostability of the enzyme
-
free papain loses 60% of its activity after 2 h at 75C, papain immobilized on the poly(glycidyl methacrylate-co-ethylene dimethylacrylate) monolith loses 10% of its activity
-
Immobilization of papain after chemical modification increases its stability and reusability in alkaline conditions. After adding of detergent powder in distilled water (final concentration of 5-25 mg/ml) to the proteolytic activity test at 45C, papains modified by 1,2,4-benzenetricarboxylic anhydride and pyromellitic anhydride are more stable. The immobilized papain modified by pyromellitic anhydride still retains about 40% of its activity in the thickest detergent. For the immobilized unmodified papain, the strong surfactant at 15 and 20 mg/ml causes a significant inhibition of 71% and 86%. At lower concentrations (0-10 mg/ml) of the immobilized papain modified by pyromellitic anhydride applied, there is a very slight decrease in the activity.
-
immobilization of the enzyme on Hiflow supercel, kaolinite clay, leaf mucilage, or starch gel leads to better thermal stability at 60-70C
-
immobilized enzyme does not lose any activity after tereatment with 6 M urea for 270 min, soluble papain loses 81% of its activity after urea treatment
-
immobilized enzyme is stable at 4C and pH 7.5 for up to 8 months, the soluble enzyme loses activity within 96 h
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in the presence of 20% (w/v) sorbitol the enzyme retains nearly 65% of its activity after heating at 75C for 40 min and it retains almost 45% of its activity after heating for 80 min, the maximum effect is seen in the case of 40% (w/v) sorbitol, where the enzyme retains nearly 80 and 70% of its activity after heating at 75C for 40 and 80 min, respectively, the enzyme also shows increased thermal stability in the presence of sucrose or xylose
-
papain exists in molten globule state at pH 2.0 and in this state protein tends to aggregate in the presence of lower concentrations of guanidine hydrochloride. Such aggregation is prevented if a low concentration of urea is also present in the buffer, in addition, stabilization of the protein is also induced
-
remains fully active after prolonged exposure to 9 M urea
-
retains activity in 8 M urea
-
solid state cysteine increases the stability of n-propanol dehydrated, immobilized enzyme in in low-water tert-butanol medium
-
temperature and guanidine hydrochloride induced unfolding transitions of papain at pH 2.0 are biphasic, implying independent and sequential unfolding of its two domains. The N-domain unfolds initially
-
the operational stability, expressed as the catalytic half-life reaches about 1 week for the enzyme immobilized on the poly(glycidyl methacrylate-co-ethylene dimethylacrylate) monolith under optimal pH and temperature conditions
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ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
acetonitrile
-
papain retains almost all its catalytic activity after 24 h of incubation in the presence of 99% (v/v) acetonitrile with a more compact conformation
dimethyl formamide
-
papain shows a complete inactivation after 24 h when exposed to those media containing 90% (v/v) dimethyl formamide because of an irreversible conformational change
Glycerol
-
in the presence of 20% (v/v) glycerol the enzyme retains nearly 80% of its activity even after heating at 75C for 40 min and it retains almost 70% of its activity even after heating for 80 min, at 30% (v/v) glycerol the enzyme retains nearly 80 and 65% of its activity after heating for 40 and 80 min, respectively at 75C
hexadecyltrimethyl ammonium bromide
-
the presence of hexadecyltrimethyl ammonium bromide allows 41% recovery of enzymatic activity of acid-unfolded papain in the presence of hexadecyltrimethyl ammonium bromide the enzyme exists as a compact intermediate with regain of native-like secondary and partial tertiary structure as well as high 8-anilino-1-naphthalene-sulfonic acid binding with the partially recovered enzymatic activity
Methanol
-
papain shows 80% loss of activity after 24 h incubation in 90% (v/v) methanol although no global conformational change and minor secondary structure rearrangements are detected
SDS
-
the presence of SDS allows 43% recovery of enzymatic activity of acid-unfolded papain, addition of 8 mM SDS results in the loss of 8-anilino-1-naphthalene-sulfonic acid binding sites exhibited by a decrease in 8-anilino-1-naphthalene-sulfonic acid fluorescence intensity, suggesting the burial of hydrophobic patches, papain at low pH and in the presence of SDS exists in a partially folded state characterized by native-like secondary structure and tertiary folds
tetrahydrofuran
-
30%, inactivation within 30 min. Sugars protect papain from tetrahydrofuran-induced inactivation in the decreasing order D-ribose, D-fructose, D-glucose, D-saccharose, D-raffinose. D-ribose at 1.6 mol per l is the most effective stabiliser. In 60% tetrahydrofuran in the presence of ribose, papain preserves about 55% of its initial activity after 2 h
Tween
-
the presence of Tween-20 allows 39% recovery of enzymatic activity of acid-unfolded papain in the presence of Tween-20, acid-unfolded papain exists as a compact intermediate with molten-globule-like characteristics
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80C, purified enzyme can be activated to at least 65% of its original activity after storage at -80C for up to 3 months
-
25C, model wine (pH 3.2, 12% ethanol), 7 days, papain from papaya latex retains 50% of activity and papain from ripe fruit retains 18% activity
-
4C, 60 days, the immobilized enzyme shows 20% loss of activity, the soluble enzyme shows 50% loss of activity
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4C, protein concentration 0.04 mM, 2 mM EDTA, pH 4.3, 3% loss of activity after 48 h, 10% loss of activity after 10 days
-
4C, several months, most stable in a reversibly inactivated form, e.g. mercuripapain
-
after 15 days, free papain retains 31% of its initial activity,whereas the immobilized papain retains 40% of initial activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
active papain is prepared from the commercial papaya latex enzyme through affinity purification on Sepharose 4B
-
ammonium sulfate precipitation
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aqueous two-phase system (40% (w/w) 15 mg/ml enzyme solution, 14.33-17.65% (w/w) PEG 6000, 14.27-14.42% (w/w) NaH2PO4/K2HPO4 and pH 5.77-6.3 at 20C). Propanol generates solid enzyme aggregates with almost 120% activity upon resolubilization through dilution of the precipitant
-
further purification of the commercial enzyme preparation from latex by gel filtration, to homogeneity
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insoluble His-tagged fusion protein is solubilized and purified by nickel chelate affinity chromatography under denaturing conditions and it is further purified by gel filtration chromatography
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purification of papain from papaya powder extracts by membranes immobilized with Reactive Red 120 or Reactive Brown 10 as dye ligands. Papain adsorption capacities for the Red 120 and Brown 10 membranes are 143.6 mg/g and 107.3 mg/g, respectively. Yields of over 80% are found for the Red 120-chitosan-nylon membrane whereas only a 50% recovery is possible using the Brown 10-CS-nylon membranes
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purification of papain using the dye affinity membrane chromatography at pH 7.05 with Tris-HCl, papain is purified 34.6fold in a single step determined by fast protein liquid chromatography
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recombinant enzyme
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recombinant papain prosegment solubilized and refolded from Escherichia coli strain BL21(DE3) inclusion bodies, to homogeneity
three kinds of affinity column for the purification of papain
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using dye ligand affinity chromatography with a cryogel column. Papain is purified 42fold in single step
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA sequence determination and analysis of the proenzyme, expression of the isolated papain prosegment in Escherichia coli strain BL21(DE3) in the insoluble fraction
expressed in Escherichia coli
-
the DNA coding for propapain is cloned and expressed as inclusion bodies at a high level in Escherichia coli BL21(DE3) transformed with two T7 promoter based pET expression vectors, pET30 Ek/LIC and pET28a+, each containing the propapain gene, recombinant propapain is expressed as an insoluble His-tagged fusion protein
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wild-type papain and D158N variant produced in a baculovirus insect cell expression system
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D158N
-
clear kinetic and pH depndence differences from the wild-type enzyme
K174R
-
mutation introduced according to thermostable homologue ervatamin C, unstable
K174R/V32S
-
mutation introduced according to thermostable homologue ervatamin C, improvement of thermal stability
K174R/V32S/G36S
-
mutation introduced according to thermostable homologue ervatamin C, improvement of thermal stability
V133A/S205E
-
change in specificity compared to wild-type enzyme
V133A/V157G/S205E
-
important decrease in activity, change in specificity compared to the wild-type enzyme
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
acid-unfolded papain in the presence of 8 mM SDS and 3.5 mM hexadecyltrimethyl ammonium bromide retains its partial tertiary structure
-
fractions containing propapain are pooled and refolded by rapid dilution into a refolding buffer at pH 8.6, the denatured protein folds best at pH 8.5 and at 4C, while pH values above 8.8 and below 7.0 are not suitable for folding, folding process requires addition of 0.5 M arginine, 1 mM glutathione, 0.1 mM GSSG and 15% glycerol
-
recombinant papain prosegment from Escherichia coli strain BL21(DE3) inclusion bodies by 2% Triton X-100, 8 M guanidinium hydrochloride, and dialysis against 50 mM phosphoric acid/NaOH buffer, pH 7.0, and centrifugation at 45000 rpm for 1 h
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
biotechnology
industry
medicine
nutrition
synthesis