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 | 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 (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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Carica papaya | P00784 | - |
- |
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 (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 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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
azocasein + H2O | - |
Carica papaya | azopeptide + peptides | - |
? |
Synonyms | Comment | Organism |
---|---|---|
papain | - |
Carica papaya |
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 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 |