An approach combining alfalfa seeds and biotechnological methods to enhance wheat bread quality: Nutritional, antioxidant, spectroscopic and sensory aspects.

Non-germinated (ASC) and germinated alfalfa seed flours (GASC) were used as wheat flour substitutes (5 and 10 g/100 g) in standard and sourdough bread production. The proximate composition, mineral, amino acid, fatty acid and phenolic profiles, and antioxidant activity of alfalfa-enriched bread were examined alongside changes in protein secondary structure and sensory acceptability. A significant increase in protein (5.83-7.35 g/100 g), total dietary fibre (1.79-4.07 g/100 g), Ca (20.31-66.83 g/100 g), Mg (22.89-44.16 g/100 g), and Fe (0.80-3.88 g/100 g) content was achieved in alfalfa-enriched bread. Alfalfa-enriched bread was also characterized by substantial quantities of lysine, aspartic acid, arginine, linolenic acid, total polyunsaturated fatty acids, total phenolics and antioxidant activity. The sensory acceptability of bread was not significantly compromised by alfalfa flour inclusion since alterations in protein secondary structure were minor according to FTIR analysis contributing to the maintenance of desirable bread quality. The alfalfa flour quantity had the strongest influence on the content of the investigated nutrients. ASC inclusion was more favourable regarding bread's antioxidant properties compared to GASC, while the usage of sourdough had a positive effect on bread's palatability compared to the standard bread-making process. Alfalfa seed flour could be an innovative ingredient to enhance the nutritional quality of wheat-based bread, while the successive germination and sourdough fermentation processes can further boost the content of particular nutrients and sensory attributes of the enriched bread.