Does the Protein Aggregation State Affect the Digestibility and Safety of Foods?

This thesis explores the complex relationship between food protein structure and digestibility. Food proteins are important nutrients that play a central role in controlling the textural properties of many foods. Processing of food proteins may alter the protein aggregate structure and digestibility. The degree of protein aggregation during food processing depends on the denaturing conditions and the presence of other food components. Sugars and lipids may contribute to protein glycation and protein cross-linking via the Maillard reaction. Furthermore, amino acid residues of food proteins may be chemically modified during processing, thereby influencing both the structure and the nutritional value of proteins. An in vitro digestibility assay was used to investigate the relationship between protein aggregate structure and protein digestibility. Raw and boiled egg whites were exposed to a wide range of conditions: pH 2 - 12, in the presence and absence of 200 mM NaCl. It was found that pH and NaCl treatment prior to in vitro digestion resulted in significantly different protein ultrastructures, but did not markedly influence protein digestibility under the tested conditions. Raw egg white was less digestible than boiled egg white under all test conditions. The inclusion of Maillard reaction partners caused protein cross-linking concurrent with a decrease in digestibility. The digestibility decreased with the reactivity of the Maillard reaction partner and with increasing heating time. Proteomic analysis, using tandem mass spectrometry, of raw and heated egg white showed an increase in hydrothermally induced amino acid modifications. In the presence of glucose and methylglyoxal, a Maillard reaction specific increase in arginine modification to hydroimidazolone was observed with increasing heating times. The observed modifications are likely to contribute to a change in the nutritional quality of egg white. Aggregation kinetics of the major egg white protein, ovalbumin, were studied by dynamic light scattering, small angle X-ray scattering, and transmission electron microscopy. Shape determination was only possible for ordered aggregates, but not for disordered aggregates. Prior to heating, ovalbumin molecules in the presence of water and glucose repelled each other in concentrated solution. The presence of NaCl shielded electrostatic repulsion, leading to early onset dimerisation and disordered aggregation upon heating. Methylglyoxal treated ovalbumin formed more ordered aggregates. The scattering of these structures was able to be fitted to cylindrical shape models showing an increase of cylinder length with time while the cylinder diameter remained near constant over 24 hours of heating. In addition, food protein derived amyloid fibril aggregates were characterised. Amyloid fibrils are a common ordered protein fold that has been linked to neurodegenerative diseases. In the recent literature, amyloid fibrils have been proposed as new functional macromolecules in proteinaceous foods because of their desirable textural properties. Food fibrils formed from whey, egg white, soy bean and kidney bean protein were tested to establish whether they are protease resistant or display toxicity to human Caco-2 cells (a model intestinal cell line). The food fibrils were compared to insulin amyloid fibrils, a well characterised amyloid system. It was shown that the food fibrils displayed some resistance towards in vitro hydrolysis and were not found to be toxic. This work contributes to the understanding of food protein aggregation and digestibility under relevant conditions. It highlights the relationship of aggregate structure and digestibility and the particular role of the Maillard reaction. Moreover, evidence is provided that food protein derived amyloid fibrils may be safe ingredients in consumables. These findings may contribute to optimising industrial food processes and creating safe new food products