Information on EC 3.4.22.31 - Ananain

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Ananas comosus

EC NUMBER
COMMENTARY
3.4.22.31
-
RECOMMENDED NAME
GeneOntology No.
Ananain
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
hydrolysis of proteins with broad specificity for peptide bonds. Best reported small molecule substrate Bz-Phe-Val-Arg-/-NHMec, but broader specificity than fruit bromelain
show the reaction diagram
From stem of pineapple plant, Ananas comosus. Differs from stem and fruit bromelains in being inhibited by chicken cystatin. In peptidase family C1 (papain family)
-
-
-
hydrolysis of proteins with broad specificity for peptide bonds. Best reported small molecule substrate Bz-Phe-Val-Arg-/-NHMec, but broader specificity than fruit bromelain
show the reaction diagram
from stem of pineapple plant, Ananas comosus,. Differs from stem and fruit bromelains in being inhibited by chicken cystatin, amino acid sequence, contains an insert between residues 170 and 174 not present in stem bromelain or papain and a hydropobic series of amino acids adjacent to His157, which possibly contributes to the different substrate and inhibitor specificities
-
hydrolysis of proteins with broad specificity for peptide bonds. Best reported small molecule substrate Bz-Phe-Val-Arg-/-NHMec, but broader specificity than fruit bromelain
show the reaction diagram
from stem of pineapple plant, Ananas comosus,. Differs from stem and fruit bromelains in being inhibited by chicken cystatin, contains an active site His residue, active site structure determination
P80884
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
hydrolysis of peptide bond
-
-
CAS REGISTRY NUMBER
COMMENTARY
119129-70-3
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
commercial preparation
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
azocasein + H2O
fragments of azocasein
show the reaction diagram
-
-
-
-
azocasein + H2O
fragments of azocasein
show the reaction diagram
-
-
-
-
azocasein + H2O
fragments of azocasein
show the reaction diagram
-
-
-
-
-
Azocoll + H2O
Hydrolyzed azocoll
show the reaction diagram
-
-
-
-
Azocoll + H2O
Hydrolyzed azocoll
show the reaction diagram
-
-
-
-
Benzoyl-Phe-Val-Arg 4-methylcoumarin 7-amide + H2O
Benzoyl-Phe-Val-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
Benzoyl-Phe-Val-Arg 4-methylcoumarin 7-amide + H2O
Benzoyl-Phe-Val-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
-
Benzoyl-Phe-Val-Arg 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Arg-Arg 4-methylcoumarin 7-amide + H2O
Benzyloxycarbonyl-Arg-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
Benzyloxycarbonyl-Arg-Arg 4-methylcoumarin 7-amide + H2O
Benzyloxycarbonyl-Arg-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
Benzyloxycarbonyl-Arg-Arg 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Gly-Phe-citrulline 4-methylcoumarin 7-amide + H2O
Benzyloxycarbonyl-Gly-Phe-citrulline + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
Benzyloxycarbonyl-Phe-Arg 4-methylcoumarin 7-amide + H2O
Benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
Benzyloxycarbonyl-Phe-Arg 4-methylcoumarin 7-amide + H2O
Benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Phe-citrulline 4-methylcoumarin 7-amide + H2O
Benzyloxycarbonyl-Phe-citrulline + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
Fibrin + H2O
Hydrolyzed fibrin
show the reaction diagram
-
-
-
-
Fibrin + H2O
Hydrolyzed fibrin
show the reaction diagram
-
-
-
-
Hemoglobin + H2O
Hydrolyzed hemoglobin
show the reaction diagram
-
-
-
-
hide powder azure + H2O
hydrolyzed hide powder azure
show the reaction diagram
-
-
-
-
hide powder azure + H2O
hydrolyzed hide powder azure
show the reaction diagram
-
-
-
-
hide powder azure + H2O
hydrolyzed hide powder azure
show the reaction diagram
-
-
-
-
-
Phosphorylase a + H2O
Hydrolyzed phosphorylase a
show the reaction diagram
-
-
-
-
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
P80884
-
-
?
pyr-Glu-Phe-Leu-4-nitroanilide + H2O
pyr-Glu-Phe-Leu + 4-nitroaniline
show the reaction diagram
P80884
-
-
?
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
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
P80884
-
-
?
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Chicken cystatin
-
strong
-
Cystatin
P80884
from chicken
-
L-3-carboxy-2,3-trans-epoxypropionylleucylamido(4-guanidino)butane
-
i.e. E-64
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
P80884
activation of the recombinant proenzyme by incubation for 30-60 min in 50 mM sodium acetate, pH 4.0, 20 mM cysteine and 4 mM EDTA at 55°C
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0484
-
benzoyl-Phe-Arg 4-methylcoumarin 7-amide
-
-
0.0131
-
benzoyl-Phe-Val-Arg 4-methylcoumarin 7-amide
-
-
1.6
-
benzoyl-Phe-Val-Arg 4-nitroanilide
-
-
0.0445
-
benzyloxycarbonyl-Arg-Arg 4-methylcoumarin 7-amide
-
-
11
-
benzyloxycarbonyl-Arg-Arg 4-nitroanilide
-
-
0.0044
-
Benzyloxycarbonyl-Gly-Phe-citrulline 4-methylcoumarin 7-amide
-
-
0.0111
-
Benzyloxycarbonyl-Phe-citrulline 4-methylcoumarin 7-amide
-
-
additional information
-
additional information
P80884
kinetics
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
63.8
-
benzoyl-Phe-Val-Arg 4-methylcoumarin 7-amide
-
-
10.9
-
benzoyl-Phe-Val-Arg 4-nitroanilide
-
-
0.321
-
benzyloxycarbonyl-Arg-Arg 4-methylcoumarin 7-amide
-
-
0.13
-
benzyloxycarbonyl-Arg-Arg 4-nitroanilide
-
-
3.54
-
Benzyloxycarbonyl-Gly-Phe-citrulline 4-methylcoumarin 7-amide
-
-
21
-
Benzyloxycarbonyl-Phe-citrulline 4-methylcoumarin 7-amide
-
-
7
-
benzyloxycarbonyl-Phe-Val-Arg 4-methylcoumarin 7-amide
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
-
additional information
-
P80884
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
5.5
P80884
recombinant mutant E35A
5.5
7
P80884
recombinant mutant E50A and recombinant wild-type enzyme
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3
9.5
P80884
-
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
P80884
assay at
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
23420
-
-
Ananas comosus, electrospray mass spectrometry
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 25000, Ananas comosus, SDS-PAGE
?
-
x * 23464, amino acid sequence determination
additional information
P80884
modeling of three-dimensional structure, enzyme lacks the extensive network of acidic residues in and around the active site
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
no glycoprotein
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20°C, stored dry, stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
by reversible pegylation
-
recombinant enzyme from Escherichia coli
P80884
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
DNA sequence determination and analysis of pre-pro-ananain, overexpression of pro-enzyme and mutants E50A and E35A in Escherichia coli BL21(DE3)
P80884
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
E35A
P80884
site-directed mutagenesis, exchange of the conserved residue functional as part of the so-called electrostatic switch, mutation leads to an increased instability of the refolded recombinant pro-enzyme, different activation conditions are required, altered pH profile
E50A
P80884
site-directed mutagenesis, exchange of the conserved residue functional as part of the so-called electrostatic switch, slightly altered pH profile and slightly enhanced activity, activation similar to the wild-type
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
refolding and activation of the overexpressed recombinant protein from inclusion bodies
P80884