Two-dimensional (2-D) IEF/SDS-PAGE is a powerful tool to get molecular “pictures” of food proteomes and monitor the processing effect(s) of a given food item on its protein profile. However, the relatively scarce diffusion of proteomic techniques at industrial and analytical level has prevented their application in this promising area so far. In this work, 2-D electrophoresis has been used to monitor the main steps of lupin-based gluten-free pasta production. To the best of our knowledge, this work is one of the rare examples of 2-D electrophoresis application to the analysis of legume seed protein components in the whole production chain of a food product. The optimised dietary exploitation of protein-rich plant sources, such as legume seeds, is the target of several research and development programs. Among the legume seeds, lupin is an interesting one for various reasons: a very high protein content (1), comparable to that of soybean, a low presence of antinutritional compounds (2), the functional properties of its components in food matrices (3) and the nutraceutical potentialities of some of its proteins (4). In this work we have studied a lupin-based pasta product, taking advantage of the presence of a single protein source in this product, being lupin used as the unique alternative to semolina, and of a previously published lupin storage protein 2-D maps (5). To this aim, three different production lots of lupin-based pasta were analysed. For each lot, samples at each critical production step, including seeds, raw materials, namely the flour and the protein concentrate, half-processed products and dry pasta, were used to generate the corresponding 2-D electrophoretic maps. The presence, integrity and constancy of proteins throughout the industrial processing have been assessed. Indeed, some differences in the protein profiles between the raw materials, i.e. lupin flour and lupin protein concentrate, were attributed to the different varieties which they arose from. On the other hand, the electrophoretic analyses showed only minor differences among the samples during the industrial processing. In particular no alteration of the covalent continuity of the main polypeptide backbones. The disulphide pattern did not change during the process, as well, and the constancy of the glycosylation pattern, as measured by the lectin Concanavalin A on the blotted maps, indicated that this molecular feature was not affected by the process too. In conclusion, this work shows that taking 2-D electrophoretic “pictures” of the polypeptides at each relevant step of a food production process can be very useful to both quality control strategies and traceability of specific protein components in force of the high resolution of the technique also in complex food matrices.

Applications of 2-D Electrophoresis and Western Blot to Analyse and Trace Proteins in Lupin based Pasta Products / J. Capraro, C. Magni, M. Duranti - In: ItPA : 2nd Annual National Conference Italian Proteomic Association[s.l] : ItPA, 2007. - pp. 101-101 (( Intervento presentato al 2. convegno Italian Proteomic Association Annual National Conference tenutosi a Aci Trezza nel 2007.

Applications of 2-D Electrophoresis and Western Blot to Analyse and Trace Proteins in Lupin based Pasta Products

J. Capraro
Primo
;
C. Magni
Secondo
;
M. Duranti
Ultimo
2007

Abstract

Two-dimensional (2-D) IEF/SDS-PAGE is a powerful tool to get molecular “pictures” of food proteomes and monitor the processing effect(s) of a given food item on its protein profile. However, the relatively scarce diffusion of proteomic techniques at industrial and analytical level has prevented their application in this promising area so far. In this work, 2-D electrophoresis has been used to monitor the main steps of lupin-based gluten-free pasta production. To the best of our knowledge, this work is one of the rare examples of 2-D electrophoresis application to the analysis of legume seed protein components in the whole production chain of a food product. The optimised dietary exploitation of protein-rich plant sources, such as legume seeds, is the target of several research and development programs. Among the legume seeds, lupin is an interesting one for various reasons: a very high protein content (1), comparable to that of soybean, a low presence of antinutritional compounds (2), the functional properties of its components in food matrices (3) and the nutraceutical potentialities of some of its proteins (4). In this work we have studied a lupin-based pasta product, taking advantage of the presence of a single protein source in this product, being lupin used as the unique alternative to semolina, and of a previously published lupin storage protein 2-D maps (5). To this aim, three different production lots of lupin-based pasta were analysed. For each lot, samples at each critical production step, including seeds, raw materials, namely the flour and the protein concentrate, half-processed products and dry pasta, were used to generate the corresponding 2-D electrophoretic maps. The presence, integrity and constancy of proteins throughout the industrial processing have been assessed. Indeed, some differences in the protein profiles between the raw materials, i.e. lupin flour and lupin protein concentrate, were attributed to the different varieties which they arose from. On the other hand, the electrophoretic analyses showed only minor differences among the samples during the industrial processing. In particular no alteration of the covalent continuity of the main polypeptide backbones. The disulphide pattern did not change during the process, as well, and the constancy of the glycosylation pattern, as measured by the lectin Concanavalin A on the blotted maps, indicated that this molecular feature was not affected by the process too. In conclusion, this work shows that taking 2-D electrophoretic “pictures” of the polypeptides at each relevant step of a food production process can be very useful to both quality control strategies and traceability of specific protein components in force of the high resolution of the technique also in complex food matrices.
Settore BIO/10 - Biochimica
2007
http://www.ictmp.ct.cnr.it/itpa2007/ATTI.pdf
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/159435
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