Techno-nutritional capabilities of sprouted clover seeds sourdough as a potent bio-preservative against sorbate-resistant fungus in fortified clean-label wheat bread

Fermented sprouted seeds are promising substrates for designing preservative-free, fortified baked goods. In the present study, predominant lactic acid bacteria (LAB) isolated from fermented sprouted clover seeds (FSC) were screened based on their potential ability to control resistant fungi towards potassium sorbate (PS). Then, controlled FSC (CFSC) containing the selected LAB isolate was used as an alternative for PS to produce clean-label enriched wheat bread. Phylogenetic evolutionary analysis led to the identification of Lacticaseibacillus rhamnosus as the selected LAB isolate with potent inhibitory effect on Aspergillus brasiliensis as the targeted PS resistant fungus. Furthermore, there was no significant difference (P > 0.05) between in situ inhibitory effects of the CFSC and PS on surface growth of the target fungus. The positive effects of sprouting/fermentation on the amino acid contents were also verified according to the HPLC-based analysis. Enriched bread with CFSC and spontaneous FSC had the highest amounts of arginine and lysine, respectively. Furthermore, sourdough fermentation led to the improvement of the crumb microstructure, increasing of antioxidant activity and reduction of phytate content compared to the control. The lowest crumb hardness (3.25 N) and the highest porosity (19.86%) were also observed in bread containing CFSC in comparison with the other samples.