Papain purification insights: Monitoring by electrophoretic approaches and MALDI-TOF peptide mass fingerprint analyses

Carlos R. Llerena-Suster; W. David Obregón; Sebastián A. Trejo; Susana R. Morcelle

doi:10.1080/00032719.2010.546022

Papain purification insights: Monitoring by electrophoretic approaches and MALDI-TOF peptide mass fingerprint analyses

Carlos R. Llerena-Suster, W. David Obregón, Sebastián A. Trejo, Susana R. Morcelle

Institute of Biotechnology and Biomedicine "Vicent Villar Palasi" (IBB)

Research output: Contribution to journal › Article › Research › peer-review

1 Citation (Scopus)

Abstract

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. © Taylor & Francis Group, LLC.

Original language	English
Pages (from-to)	2124-2137
Journal	Analytical Letters
Volume	44
Issue number	12
DOIs	https://doi.org/10.1080/00032719.2010.546022
Publication status	Published - 1 Aug 2011

Keywords

Caricain
Chymopapain
Glycyl endopeptidase
MALDI-TOF peptide mass fingerprint
Papain

Access to Document

10.1080/00032719.2010.546022

Cite this

@article{d08f247c1b524f8ab8f815eceb2f5a34,

title = "Papain purification insights: Monitoring by electrophoretic approaches and MALDI-TOF peptide mass fingerprint analyses",

abstract = "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. {\textcopyright} Taylor & Francis Group, LLC.",

keywords = "Caricain, Chymopapain, Glycyl endopeptidase, MALDI-TOF peptide mass fingerprint, Papain",

author = "Llerena-Suster, {Carlos R.} and Obreg{\'o}n, {W. David} and Trejo, {Sebasti{\'a}n A.} and Morcelle, {Susana R.}",

year = "2011",

month = aug,

day = "1",

doi = "10.1080/00032719.2010.546022",

language = "English",

volume = "44",

pages = "2124--2137",

number = "12",

}

TY - JOUR

T1 - Papain purification insights: Monitoring by electrophoretic approaches and MALDI-TOF peptide mass fingerprint analyses

AU - Llerena-Suster, Carlos R.

AU - Obregón, W. David

AU - Trejo, Sebastián A.

AU - Morcelle, Susana R.

PY - 2011/8/1

Y1 - 2011/8/1

N2 - 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. © Taylor & Francis Group, LLC.

AB - 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. © Taylor & Francis Group, LLC.

KW - Caricain

KW - Chymopapain

KW - Glycyl endopeptidase

KW - MALDI-TOF peptide mass fingerprint

KW - Papain

U2 - 10.1080/00032719.2010.546022

DO - 10.1080/00032719.2010.546022

M3 - Article

SN - 0003-2719

VL - 44

SP - 2124

EP - 2137

JO - Analytical Letters

JF - Analytical Letters

IS - 12

ER -

Papain purification insights: Monitoring by electrophoretic approaches and MALDI-TOF peptide mass fingerprint analyses

Abstract

Keywords

Access to Document

Fingerprint

Cite this