Hi all,
I would like to know how can I estimate the activity of the enzyme, papain, which is used in reactions as crude latex extract of papaya.
Please share some procedures if anybody having any.
Thanks in advance.
Faslu
Papain activity may be determined either by measuring the rate of digestion of proteins, or by following the rate of hydrolysis of small molecular weight synthetic substrates such as esters or amide derivatives of amino acids.
Papain is a protease, and the classical protease assay is to incubate the sample with a protein modified with coloured (azocasein, doi:10.1016/B978-0-12-091302-2.50097-9) or fluorescent markers. The protein is then precipitated by TCA and the coloured amino acids in the supernatant are detected. Modern versions of this assay use two resonant fluorescent markers on the protein, as the protein is digested fluorescence resonance energy transfer (doi: 10.1002/andp.19484370105) is reduced and the fluorescence of the donor increases. Thus, the assay works without separation of substrate and product. Molecular Probes (now part of Thermo Fisher) has developed such assays, IIRC.
Take a look at the MilliporeSigma Website for the specific assay of Papain
Papain activity may be determined either by measuring the rate of digestion of proteins, or by following the rate of hydrolysis of small molecular weight synthetic substrates such as esters or amide derivatives of amino acids.
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts