Activating Compound | Comment | Organism | Structure |
---|---|---|---|
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 (Comment) | Organism |
---|---|
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 | Comment | Organism |
---|---|---|
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 |
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 | Comment | Organism | Structure |
---|---|---|---|
cystatin | from chicken | Ananas comosus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics | Ananas comosus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
protein + H2O | Ananas comosus | - |
peptides | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Ananas comosus | P80884 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme from Escherichia coli | Ananas comosus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
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 (Comment) | Organism |
---|---|
refolding and activation of the overexpressed recombinant protein from inclusion bodies | Ananas comosus |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
stem | - |
Ananas comosus | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Ananas comosus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
protein + H2O | - |
Ananas comosus | peptides | - |
? | |
pyr-Glu-Phe-Leu-4-nitroanilide + H2O | - |
Ananas comosus | pyr-Glu-Phe-Leu + 4-nitroaniline | - |
? |
Subunits | Comment | Organism |
---|---|---|
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] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Ananas comosus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | 5.5 | recombinant mutant E35A | Ananas comosus |
5.5 | 7 | recombinant mutant E50A and recombinant wild-type enzyme | Ananas comosus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
3 | 9.5 | - |
Ananas comosus |