Literature summary extracted from
Carter, C.E.; Marriage, H.; Goodenough, P.W.
Mutagenesis and kinetic studies of a plant cysteine proteinase with an unusual arrangement of acidic amino acids in and around the active site (2000), Biochemistry, 39, 11005-11013.
Activating Compound
EC Number |
Activating Compound |
Comment |
Organism |
Structure |
---|
3.4.22.31 |
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 |
Ananas comosus |
|
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
3.4.22.31 |
DNA sequence determination and analysis of pre-pro-ananain, overexpression of pro-enzyme and mutants E50A and E35A in Escherichia coli BL21(DE3) |
Ananas comosus |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
3.4.22.31 |
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 |
Ananas comosus |
3.4.22.31 |
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 |
Ananas comosus |
Inhibitors
EC Number |
Inhibitors |
Comment |
Organism |
Structure |
---|
3.4.22.31 |
cystatin |
from chicken |
Ananas comosus |
|
KM Value [mM]
EC Number |
KM Value [mM] |
KM Value Maximum [mM] |
Substrate |
Comment |
Organism |
Structure |
---|
3.4.22.31 |
additional information |
- |
additional information |
kinetics |
Ananas comosus |
|
Natural Substrates/ Products (Substrates)
EC Number |
Natural Substrates |
Organism |
Comment (Nat. Sub.) |
Natural Products |
Comment (Nat. Pro.) |
Rev. |
Reac. |
---|
3.4.22.31 |
protein + H2O |
Ananas comosus |
- |
peptides |
- |
? |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
3.4.22.31 |
Ananas comosus |
P80884 |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
3.4.22.31 |
recombinant enzyme from Escherichia coli |
Ananas comosus |
Reaction
EC Number |
Reaction |
Comment |
Organism |
Reaction ID |
---|
3.4.22.31 |
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 |
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 |
Ananas comosus |
|
Renatured (Commentary)
EC Number |
Renatured (Comment) |
Organism |
---|
3.4.22.31 |
refolding and activation of the overexpressed recombinant protein from inclusion bodies |
Ananas comosus |
Source Tissue
EC Number |
Source Tissue |
Comment |
Organism |
Textmining |
---|
3.4.22.31 |
stem |
- |
Ananas comosus |
- |
Specific Activity [micromol/min/mg]
EC Number |
Specific Activity Minimum [µmol/min/mg] |
Specific Activity Maximum [µmol/min/mg] |
Comment |
Organism |
---|
3.4.22.31 |
additional information |
- |
- |
Ananas comosus |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
3.4.22.31 |
protein + H2O |
- |
Ananas comosus |
peptides |
- |
? |
|
3.4.22.31 |
pyr-Glu-Phe-Leu-4-nitroanilide + H2O |
- |
Ananas comosus |
pyr-Glu-Phe-Leu + 4-nitroaniline |
- |
? |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
3.4.22.31 |
More |
modeling of three-dimensional structure, enzyme lacks the extensive network of acidic residues in and around the active site |
Ananas comosus |
Temperature Optimum [°C]
EC Number |
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
Organism |
---|
3.4.22.31 |
25 |
- |
assay at |
Ananas comosus |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
3.4.22.31 |
5 |
5.5 |
recombinant mutant E35A |
Ananas comosus |
3.4.22.31 |
5.5 |
7 |
recombinant mutant E50A and recombinant wild-type enzyme |
Ananas comosus |
pH Range
EC Number |
pH Minimum |
pH Maximum |
Comment |
Organism |
---|
3.4.22.31 |
3 |
9.5 |
- |
Ananas comosus |