African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12306

Full Length Research Paper

Critical evaluation of proteomic protocols for passion fruit (Passiflora edulis Sims) leaves, a crop with juice market benefits

Viviane A. Perdizio1, Olga L. T. Machado2, Jucelia S. Araujo2, Antonia E. A. Oliveira2, Leandro R. Monteiro3, Monique N. Costa4, Andre T. Ferreira4, Jonas Perales4 and Tania Jacinto1*
1Laboratorio de Biotecnologia, Centro de Biociencias e Biotecnologia, Universidade Estadual do Norte Fluminense, 28013-600, Campos dos Goytacazes, RJ, Brazil. 2Laboratorio de Quimica de Funcao de Proteinas e Peptideos, Centro de Biociencias e Biotecnologia, Universidade Estadual do Norte Fluminense, 28013-600, Campos dos Goytacazes, RJ, Brazil. 3Laboratorio de Ciencias Ambientais, Centro de Biociencias e Biotecnologia, Universidade Estadual do Norte Fluminense, 28013-600, Campos dos Goytacazes, RJ, Brazil. 4Laboratorio de Toxinologia, Instituto Oswaldo Cruz, Fundacao Oswaldo Cruz, 21040-360, Rio de Janeiro, RJ, Brazil.
Email: [email protected]

  •  Received: 30 October 2014
  •  Accepted: 11 May 2015
  •  Published: 13 May 2015

Abstract

Passion fruit grows practically all over Brazilian territory; its production is largely destined to juice industry and expanding to overseas markets. The suitability of four protein extraction protocols for plant proteome was investigated to determine the best choice for studies concerning passion fruit leaf proteins. Trichloroacetic acid (TCA)/acetone extraction; isoelectric focusing (IEF) buffer extraction; phenol (Phe) extraction and Phe-SDS extraction were tested. The Phe method produced the best results, showing higher reproducibility of resolved protein spots and clearer 2D gel background staining. In comparison, the Phe-SDS method presented fewer spots and lower reproducibility. The TCA/acetone method produced the fewest identifiable spots and the IEF buffer produced the poorest results, displaying fewer reproducibly detected spots, more vertical streaks and darker 2D staining. Selected spots, obtained with Phe method, were identified by spectrometric analysis (MALDI-TOF-TOF) to exemplify the viability to perform more comprehensive proteomic studies with passion fruit leaves and, therefore increase information about stress-related and developmental responses in this fruit crop.
 
Key words: Passion fruit, proteomic, protein extraction, juice industry.