Purification of plant complex protein extracts in non-denaturing conditions by in-solution isoelectric focusing |
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Authors: | R.A. Ferreira S. Martins-Dias |
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Affiliation: | Centre for Natural Resources and the Environment (CERENA), Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal |
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Abstract: | An alternative approach for plant complex protein extracts pre-purification by in-solution isoelectric focusing in non-denaturing conditions is presented. The separation of biologically active proteins, in narrow ranges of isoelectric point (pI) was obtained by a modified OFFGEL electrophoresis. Two different water-soluble protein extracts from Phragmites leaves were fractionated into 24 fractions within a 3–10 pI range at 10 °C in the absence of denaturing/reducing agents. One-dimensional electrophoretic analysis revealed different protein distribution patterns and the effective fractionation of both protein extracts. Peroxidase activity of each fraction confirmed that proteins remained active and pre-purification occurred. Biological triplicates assured the needed reproducibility. |
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Keywords: | In-solution IEF Non-denaturing conditions OFFGEL electrophoresis pI Phragmites sp. Plant protein purification |
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