Chia (Salvia hispanica L.) shoot cultures as a scalable source of bioactive metabolites: from agar cultures to bioreactor systems

This study presents a comprehensive biotechnological characterization of Salvia hispanica (chia) shoot cultures as a scalable source of antioxidant and antimicrobial compounds. Cultures were established in agar, agitated, and temporary-immersion systems (TIS, Plantform bioreactors), with phenylalanine (Phe) supplementation to enhance secondary metabolism. Agitated cultures with 3 mmol/L Phe showed the highest rosmarinic acid production (8621.77 mg/100 g DW), over 40-fold higher than control cultures. LC–Q-Orbitrap HRMS analysis identified 127 metabolites, including phenolic acids, flavonoids, and terpenoids, revealing significant shifts in phenolic biosynthesis upon Phe treatment. Bioreactor cultures achieved the highest biomass yield (8.43-fold increase) and elevated levels of vitamins C and E. Extracts obtained from agitated and bioreactor micorshoot cultures exhibited antibacterial and antifungal activity. The highest antibacterial activity was observed for extracts from agitated cultures supplemented with 3 mM Phe, particularly against Staphylococcus aureus and Micrococcus luteus , whereas extracts from bioreactor cultures generally showed lower antimicrobial activity. These findings demonstrate that combining precursor feeding, culture optimization, and metabolomics enables efficient production of rosmarinic acid-rich biomass with potential applications in food, pharmaceutical, and cosmetic industries.