Simplified electrophoretic assay for human salivary alpha-amylase inhibitor detection in cereal seed flours.
In: Journal of agricultural and food chemistry, Jg. 55 (2007-05-30), Heft 11, S. 4334-9
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Zugriff:
Alpha-amylase inhibitors are antinutritional proteins largely found in cereal seeds. An in-gel assay was developed that allowed the rapid screening of these compounds in complex seed extracts. The assay was based on the electrophoretic separation of the extract proteins on starch-containing gels, followed by the detection of alpha-amylase-inhibiting proteins after incubation of the gel in an alpha-amylase solution; inhibitors were revealed by a staining method based on iodine binding to nondigested starch. The one-dimensional method can be useful to test inhibitory activity of purified proteins or to assay fractions recovered during a purification procedure. A two-dimensional (IEF x PAGE) non-denaturing system with second-dimension separation on starch-PAGE was also developed; the technique allowed the screening of complex protein mixtures for multiple inhibitory proteins. The newly developed assay method was used to test the presence of inhibitory activity in a crude extract from wheat flour, and it was validated by comparing in-gel and in-solution assays of commercially available alpha-amylase inhibitors.
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Simplified electrophoretic assay for human salivary alpha-amylase inhibitor detection in cereal seed flours.
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Autor/in / Beteiligte Person: | Fontanini, D ; Capocchi, A ; Saviozzi, F ; Galleschi, L |
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Zeitschrift: | Journal of agricultural and food chemistry, Jg. 55 (2007-05-30), Heft 11, S. 4334-9 |
Veröffentlichung: | Washington, American Chemical Society., 2007 |
Medientyp: | academicJournal |
ISSN: | 0021-8561 (print) |
DOI: | 10.1021/jf070122b |
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