AGROALNEXT Project: Development of new functional and probiotic products obtained through the valorization of Brassica by-products. Industrial optimization of shelf life using emerging techniques and application in the food and cosmetics industries.

The beverage showed cytotoxic effects on colon cancer cells, inducing Ca2+ overload, free radical production, and mitochondrial stress. Ferroptosis and endoplasmic reticulum stress occurred through lipid oxidation and UPR activation. Antioxidant, autophagy, and proteasome pathways acted in response to the stress. The beverage, made with broccoli byproducts (leaves), showed great potential as an anticancer functional food. The resulting beverage showed high stability and bioaccessibility of sulforaphane, polyphenols, and indoles after in vitro digestion. Fermentation modulated the bioactive compounds, favoring more bioavailable forms for humans, enhancing the functional value of broccoli, and improving the release and absorption of its phytochemicals, such as sulforaphane. The beverage retains bioactive compounds that could contribute to antioxidant and anticancer effects in the gut. Fermentation improved the extraction of bioactive compounds from broccoli byproducts. The process was optimized at 27.77 °C and 0.1 g/L (w/v) for 4 days. The fermented beverage made with broccoli leaves exhibited high levels of sulforaphane (4.38–8.82 mg/L). During preservation, the use of high hydrostatic pressure (200 MPa, 10 min) stabilized the phytochemicals and lactic acid bacteria (LAB) and maintained a 40% antidiabetic potential.