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Literature summary for 3.4.22.2 extracted from

  • Choudhury, D.; Roy, S.; Chakrabarti, C.; Biswas, S.; Dattagupta, J.K.
    Production and recovery of recombinant propapain with high yield (2009), Phytochemistry, 70, 465-472.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
cysteine the requirement for reducing conditions during the activation process is investigated with 20 mM DTT or 20 mM cysteine, propapain does not convert to papain even after prolonged incubation with DTT, whereas in presence of 20 mM cysteine, the activation occurs between 30 and 40 min at 50°C Carica papaya

Application

Application Comment Organism
industry papain has many uses and functions in a variety of industries: clarifying beer, meat tenderization, preservation of spices, contact lens cleaners, detergents, pet food palatability, digestive aids, blood stain remover, blood coagulant and cosmetics Carica papaya
medicine papain is widely used for many medical and para-medical purposes such as to assist protein digestion in chronic dyspepsia, gastric fermentation, gastritis, removal of necrotic tissue, preparation of tyrosine derivatives for the treatment of Parkinsonism, preparation of tetanus vaccines, skin cleansing agents and acne treatment Carica papaya

Cloned(Commentary)

Cloned (Comment) Organism
the DNA coding for propapain is cloned and expressed as inclusion bodies at a high level in Escherichia coli BL21(DE3) transformed with two T7 promoter based pET expression vectors, pET30 Ek/LIC and pET28a+, each containing the propapain gene, recombinant propapain is expressed as an insoluble His-tagged fusion protein Carica papaya

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
additional information
-
during processing of propapain to papain, 2 intermediates are observed in the range of 30-38 kDa at 20 min incubation at 50°C indicating that processing of propapain to papain occurs in a stepwise manner Carica papaya
42000
-
propapain containing respective fusion-tag sequence, determined by SDS-PAGE Carica papaya

Organism

Organism UniProt Comment Textmining
Carica papaya P00784
-
-

Purification (Commentary)

Purification (Comment) Organism
insoluble His-tagged fusion protein is solubilized and purified by nickel chelate affinity chromatography under denaturing conditions and it is further purified by gel filtration chromatography Carica papaya

Renatured (Commentary)

Renatured (Comment) Organism
fractions containing propapain are pooled and refolded by rapid dilution into a refolding buffer at pH 8.6, the denatured protein folds best at pH 8.5 and at 4°C, while pH values above 8.8 and below 7.0 are not suitable for folding, folding process requires addition of 0.5 M arginine, 1 mM glutathione, 0.1 mM GSSG and 15% glycerol Carica papaya

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
125.9
-
recombinant papain, in 100 mM sodium acetate and 2 mM EDTA at pH 6.5 Carica papaya
221.7
-
commercial papain, in 100 mM sodium acetate and 2 mM EDTA at pH 6.5 Carica papaya

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
azocasein + H2O
-
Carica papaya azopeptide + peptides
-
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Synonyms

Synonyms Comment Organism
papain
-
Carica papaya

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
additional information
-
processing of propapain to mature papain is triggered by low pH and is stimulated by high temperatures Carica papaya

pH Range

pH Minimum pH Maximum Comment Organism
3.8 4.5 conditions for in vitro activation of the refolded propapain to mature papain are determined, the highest rates of activation are observed between pH 3.8 and 4.5, while the rates are greatly reduce at pH higher than 6 Carica papaya