Papain Purification Insights: Monitoring by Electrophoretic Approaches and MALDI-TOF Peptide Mass Fingerprint Analyses

Papain was purified from dried Carica papaya latex by fractioned salt precipitation in presence of sodium tetrathionate to preserve enzymatic activity. Purification was followed by different electrophoretic methods. Identification of the purified product was afforded by submitting the peptides obtai...

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Bibliographic Details
Published in:Analytical letters 2011-08, Vol.44 (12), p.2124-2137
Main Authors: Llerena-Suster, Carlos R., Obregón, W. David, Trejo, Sebastián A., Morcelle, Susana R.
Format: Article
Language:English
Subjects:
Analytical chemistry
Caricain
Chemistry
Chymopapain
Digestion
Exact sciences and technology
Glycyl endopeptidase
Latex
MALDI-TOF peptide mass fingerprint
Monitoring
Papain
Papayas
Peptides
Precipitation
Purification
Spectrometric and optical methods
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Summary:Papain was purified from dried Carica papaya latex by fractioned salt precipitation in presence of sodium tetrathionate to preserve enzymatic activity. Purification was followed by different electrophoretic methods. Identification of the purified product was afforded by submitting the peptides obtained by tryptic digestion of papain to matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/TOF MS) analysis. Comparison of the peptide masses analyzed by peptide mass fingerprinting (PMF) MALDI-TOF and those obtained by theoretical tryptic digestion, revealed the presence of some peptides belonging the other three endopeptidases contained in papaya latex (very similar to papain in molecular weight and pI) in the purified fraction of papain. The PMF by MALDI-TOF could be applied as a method to follow papain purification.
ISSN:0003-2719
1532-236X
DOI:10.1080/00032719.2010.546022