Evalution of 1D and 2D gel-based proteome from fresh leaf of Moringa oleifera

Abstract

Moringa species is one of the most useful trees in tropics and subtropics of Asia and Africa. Almost all parts of Moringa are edible and have been consumed as vegetable. Furthermore, due to low amounts of proteins in plant tissues the optimization of protein extraction and to establish a robust protocol for two-dimensional gel electrophoresis (2-DE) along with downstream processing are important step. Moreover, the problem of unidentified protein and protein spots from the leaves of M. oleifera can be overcome by applying two dimension gel electrophoresis. In this study, two conventional methods and two kits were evaluated on fresh leaf of Moringa oleifera to distinguish the most suitable protein extraction method. Besides, the protein pattern and spots distribution were determined from 1D and 2D proteomics respectively. From SDS-PAGE it was shown that protein extraction using QB method was the best method, resulting in more protein bands which allowed detection of 24 bands approximately. In addition, from two dimensional gel electrophoresis it was revealed that method 2 (2000 V) was the best method compared to method 1 (1000 V) as it gives better spot distribution and reproducible where the protein spots were distributed mainly within the pH range of 4-7.