Amino Acids, Peptides and Proteins - Dévényi T., Gergely J. 1976

Hydrolysis of proteins and peptides
Hydrolysis of proteins and peptides by papain

Principle of the method. Papain, which lacks pronounced Specificity, hydrolyzes Proteins AND Peptides into small fragments.

Applications. Structural Analysis of proteins and peptides.

PROCEDURE

1. Activation of papain. 10 mg of commercially available papain is dissolved in 1 mL of 1% NaCl and incubated at 37°C for 1 h. Immediately after this Treatment, the enzyme is ready for use.

2. Papain Hydrolysis. Papain is added to a 1% solution of the substrate (protein) to be hydrolyzed to a final concentration of 0.02%, and the pH is adjusted to 7.0 with 1.0 N NaOH. The mixture is incubated in an autotitrator at room Temperature under a nitrogen atmosphere, with slow stirring on a magnetic stirrer for 20–24 h. The reaction is stopped by boiling, and the reaction mixture is filtered and lyophilized.

NOTE

Papain is a non-specific enzyme; it cleaves various peptide bonds, such as Glu-Gln, Cys SO3H-Ser, Gln-Leu, Glu-Asn, etc.



Last update: 06/08/2026

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