Cysteine proteinases present in pineapple plants are phytotherapeutical agents with anti-edematous, anti-inflammatory, anti-thrombotic and fibrinolytic activities. Active components involved have been only partially identified as bromelain, the major proteinase in pineapple fruits.In this work, pineapple fruit extracts were analysed by 2-D zymography. Clear spots, corresponding to enzymatic activities, were excised, digested with trypsin and submitted to MALDI-ToF mass spectrometry for enzyme identification. The most representative enzymes were identified as bromelains, including their isoforms and their post-translational modifications. The novelty of the present study is the identification of proteolytic activities by means of direct MALDI-ToF MS analysis of the zymographic spots. Enzymes were identified without the need for 2-D gel electrophoresis or purification.2-D zymography can offer not only the complete map of the enzymes present in a biological sample or in a food matrix, but it can allow also their direct identification including their isotypes. © 2010 Elsevier Ltd.

Analysis of pineapple [Ananas comosus (L.) Merr.] fruit proteinases by 2-D zymography and direct identification of the major zymographic spots by mass spectrometry

Santamaria M.;Riccio P.
2010-01-01

Abstract

Cysteine proteinases present in pineapple plants are phytotherapeutical agents with anti-edematous, anti-inflammatory, anti-thrombotic and fibrinolytic activities. Active components involved have been only partially identified as bromelain, the major proteinase in pineapple fruits.In this work, pineapple fruit extracts were analysed by 2-D zymography. Clear spots, corresponding to enzymatic activities, were excised, digested with trypsin and submitted to MALDI-ToF mass spectrometry for enzyme identification. The most representative enzymes were identified as bromelains, including their isoforms and their post-translational modifications. The novelty of the present study is the identification of proteolytic activities by means of direct MALDI-ToF MS analysis of the zymographic spots. Enzymes were identified without the need for 2-D gel electrophoresis or purification.2-D zymography can offer not only the complete map of the enzymes present in a biological sample or in a food matrix, but it can allow also their direct identification including their isotypes. © 2010 Elsevier Ltd.
File in questo prodotto:
Non ci sono file associati a questo prodotto.

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11586/401502
 Attenzione

Attenzione! I dati visualizzati non sono stati sottoposti a validazione da parte dell'ateneo

Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus 27
  • ???jsp.display-item.citation.isi??? 20
social impact