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Literature summary for 3.4.22.31 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.
    View publication on PubMed
Search for more literature for 3.4.22.31

Activating Compound

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(Commentary)

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

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

Inhibitors Comment Organism Structure
cystatin from chicken Ananas comosus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information kinetics Ananas comosus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
protein + H2O Ananas comosus
-
 peptides
-
 ?

Organism

Organism UniProt Comment Textmining
Ananas comosus P80884
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme from Escherichia coli Ananas comosus

Reaction

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
a

Renatured (Commentary)

Renatured (Comment) Organism
refolding and activation of the overexpressed recombinant protein from inclusion bodies Ananas comosus

Source Tissue

Source Tissue Comment Organism Textmining
stem
-
 Ananas comosus
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
 Ananas comosus

Substrates and Products (Substrate)

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

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Ananas comosus

pH Optimum

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 Range

pH Minimum pH Maximum Comment Organism
3 9.5
-
 Ananas comosus