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

Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasi" (IBB)

Producció científica: Contribució a revista › Article › Recerca › Avaluat per experts

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Resum

La papaïna es va purificar del làtex sec de Carica papaya mitjançant precipitació fraccionada en presència de tetrationat sòdic per preservar l'activitat enzimàtica. La purificació va ser seguida per diferents mètodes electroforètics. La identificació del producte purificat es va aconseguir enviant els pèptids obtinguts per digestió tríptica de la papaïna a anàlisi per desorció/ionització-temps de vol (MALDI-TOF/TOF MS) assistida per matriu. La comparació de les masses dels pèptids analitzats mitjançant l'anàlisi d'empremta de massa del pèptid (PMF) MALDI-TOF i els obtinguts per digestió teòrica, va revelar la presència d'alguns pèptids que pertanyien a altres tres endopeptidases contingudes en el làtex de papaia (molt similars a la papaïna en pes molecular i pI) en la fracció purificada de la papaïna. L'anàlisi mitjançant PMF per MALDI-TOF es va poder aplicar com un mètode per seguir la purificació de la papaïna. © Taylor & Francis Group, LLC.

Idioma original	Anglès
Pàgines (de-a)	2124-2137
Revista	Analytical Letters
Volum	44
Número	12
DOIs	https://doi.org/10.1080/00032719.2010.546022
Estat de la publicació	Publicada - 1 d’ag. 2011

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10.1080/00032719.2010.546022

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@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",

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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

UR - https://www.scopus.com/pages/publications/80051712369

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

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