Biochemical composition and functional application of enzyme-assisted extracts from leaves of Cannabis sativa L.

Enzyme-assisted extraction (EAE) has recently gained attention for its effectiveness and eco-friendly manner of extracting valuable compounds, enhancing their nutritional value at a lower cost. Cannabis sativa L. is a widely distributed and one of the oldest plant species in the Cannabaceae family. However, the plant leaves have not been widely studied for their biochemical and functional applications. This study aimed to investigate the biochemical composition, antioxidant activity, and prebiotic potential of EAE from C. sativa leaves. Extracts were obtained under optimal conditions (3:15 h of extraction, temperature 45°C, pH 4.9, and 1% enzyme (v/w of dry matter)). The extractions were performed using Viscozyme L, Cellulase enzymes, and control without enzyme. The antioxidant activity of C. sativa leaf extracts was assessed using ABTS•+, FRAP, and ORAC assays, while the total phenolic content was obtained using the Folin-Ciocalteu method. The prebiotic potential of C. sativa leaf extracts was evaluated in an in vitro screening model using probiotic strains Lactobacillus casei 01 and Bifidobacterium animalis BLC. Extracts were tested at 5% (v/v) and 50% (v/v) concentrations. The results indicated that the yield after EAE with Viscozyme L enzyme mixture increased by up to 44% compared with EAE with Cellulase enzyme and control. Moreover, the highest antioxidant activity was established in EAE with Viscozyme L enzyme extract within all tested assays. Furthermore, all tested extracts were established to possess prebiotic potential with tested probiotic strains, but the greatest results were obtained in EAE with Viscozyme L enzyme mixture extract. In conclusion, EAE may be an effective and sustainable method for biorefining bioactive compounds, potentially applicable in high-value functional foods and nutraceuticals.