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Evaluation of protein extraction methods to obtain protein concentrate from cassava leaf REA
Coldebella,Priscila F.; Gomes,Simone D.; Evarini,Janete A.; Cereda,Marney P.; Coelho,Sílvia R. M.; Coldebella,Anderson.
The cassava leaf, waste generated in the harvest of the roots, is characterized by high content of protein, vitamins and minerals; however, its use is limited due to the high fiber content and antinutritional substances, which can be removed by obtaining protein concentrates. In this context, the objective of this study was to evaluate protein extraction processes, aiming the use of cassava leaves (Manihot esculenta Crantz) as an alternative protein. Four methods were tested: 1) Coagulation of Proteins by Lowering the Temperature, 2) Extraction by Isoelectric Precipitation, 3) Solubilization of Proteins and 4) Fermentation of Filter Leaf Juice. To obtain the concentrates, the use of fresh or dried leaves and extraction in one or two steps were also...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Exploitation; Agricultural waste; Protein extraction.
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-69162013000600015
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Method optimization for proteomic analysis of soybean leaf: improvements in identification of new and low-abundance proteins Genet. Mol. Biol.
Mesquita,Rosilene Oliveira; Soares,Eduardo de Almeida; Barros,Everaldo Gonçalves de; Loureiro,Marcelo Ehlers.
The most critical step in any proteomic study is protein extraction and sample preparation. Better solubilization increases the separation and resolution of gels, allowing identification of a higher number of proteins and more accurate quantitation of differences in gene expression. Despite the existence of published results for the optimization of proteomic analyses of soybean seeds, no comparable data are available for proteomic studies of soybean leaf tissue. In this work we have tested the effects of modification of a TCA-acetone method on the resolution of 2-DE gels of leaves and roots of soybean. Better focusing was obtained when both mercaptoethanol and dithiothreitol were used in the extraction buffer simultaneously. Increasing the number of washes...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Proteomics; Glycine max; 2-DE; Protein extraction; Leaf proteome.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000200017
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A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots Genet. Mol. Biol.
Rodrigues,Elisete Pains; Torres,Adalgisa Ribeiro; Batista,Jesiane Stefânia da Silva; Huergo,Luciano; Hungria,Mariangela.
Sample preparation is a critical step in two-dimensional gel electrophoresis (2-DE) of plant tissues. Here we describe a phenol/SDS procedure that, although greatly simplified, produced well-resolved and reproducible 2-DE profiles of protein extracts from soybean [Glycine max (L.) Merril] roots. Extractions were made in three replicates using both the original and simplified procedure. To evaluate the quality of the extracted proteins, ten spots were randomly selected and identified by mass spectrometry (MS). The 2-DE gels were equally well resolved, with no streaks or smears, and no significant differences were observed in protein yield, reproducibility, resolution or number of spots. Mass spectra of the ten selected spots were compared with database...
Tipo: Info:eu-repo/semantics/article Palavras-chave: 2-DE; Protein extraction; Proteomics; Soybean roots; Symbiosis.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000200016
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