Information on EC 3.4.22.31 - Ananain

New: Word Map on EC 3.4.22.31
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:
Search Reference ID:
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 hide
3.4.22.31
-
RECOMMENDED NAME
GeneOntology No.
Ananain
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
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
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
CAS REGISTRY NUMBER
COMMENTARY hide
119129-70-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
azocasein + H2O
fragments of azocasein
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-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-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-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
Hemoglobin + H2O
Hydrolyzed hemoglobin
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
pyr-Glu-Phe-Leu-4-nitroanilide + H2O
pyr-Glu-Phe-Leu + 4-nitroaniline
show the reaction diagram
-
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
protein + H2O
peptides
show the reaction diagram
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
chicken cystatin
-
strong
-
Cystatin
from chicken
-
L-3-carboxy-2,3-trans-epoxypropionylleucylamido(4-guanidino)butane
-
i.e. E-64
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
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]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
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
kinetics
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
63.8
benzoyl-Phe-Val-Arg 4-methylcoumarin 7-amide
Ananas comosus
-
-
10.9
benzoyl-Phe-Val-Arg 4-nitroanilide
Ananas comosus
-
-
0.321
benzyloxycarbonyl-Arg-Arg 4-methylcoumarin 7-amide
Ananas comosus
-
-
0.13
benzyloxycarbonyl-Arg-Arg 4-nitroanilide
Ananas comosus
-
-
3.54
Benzyloxycarbonyl-Gly-Phe-citrulline 4-methylcoumarin 7-amide
Ananas comosus
-
-
21
Benzyloxycarbonyl-Phe-citrulline 4-methylcoumarin 7-amide
Ananas comosus
-
-
7
benzyloxycarbonyl-Phe-Val-Arg 4-methylcoumarin 7-amide
Ananas comosus
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 5.5
recombinant mutant E35A
5.5 - 7
recombinant mutant E50A and recombinant wild-type enzyme
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
23420
-
Ananas comosus, electrospray mass spectrometry
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
modeling of three-dimensional structure, enzyme lacks the extensive network of acidic residues in and around the active site
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
no glycoprotein
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, stored dry, stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by reversible pegylation
-
recombinant enzyme from Escherichia coli
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA sequence determination and analysis of pre-pro-ananain, overexpression of pro-enzyme and mutants E50A and E35A in Escherichia coli BL21(DE3)
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E35A
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
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
LITERATURE
refolding and activation of the overexpressed recombinant protein from inclusion bodies