Abstract The pseudo-fruit of Rosa canina L. represents a valuable resource of bioactive compounds known for their antioxidant, anti-inflammatory, antimicrobial, anti-diabetic, anti-aging, and anticancer properties. In this work, the enzyme-assisted extraction (EAE) of bioactive compounds from pseudo-fruit of Rosa canina L. was developed and optimized using four hydrolytic enzyme preparations: cellulolytic (Cellic® CTec3 HS), pectinolytic (Pectinex® Ultra Color), hemicellulolytic (Viscoferm®), and proteolytic (Neutrase®). A six-factor Box–Behnken Design (BBD) was employed to systematically evaluate the effects of enzyme loadings-Cellic® CTec3 (0.1–0.3% v/v), Pectinex® Ultra Color (0.1–0.3% v/v), Viscoferm® (0.2–0.6% v/v), and Neutrase® (0.1–0.2% v/v)-as well as agitation speed (300–1300 rpm) and extraction time (60–120 min) on the overall process. The extracts were characterized in terms of their performance in total phenolic compounds (TPC) by the Folin Ciocalteu method, in total flavonoid content (TFC) by the aluminum chloride colorimetric method, and in antioxidant activity by the DPPH free radical scavenging capacity. The optimum extract was evaluated for its biological activities, anti-aging (inhibition of tyrosinase activity), antidiabetic (inhibition of α-amylase activity), and antibacterial activity against Escherichia coli . The optimal conditions were determined by the Derringer desirability function for 0.1%v/v Cellic® CTec3 HS, 0.1%v/v Pectinex® Ultra Color, 0.21%v/v Viscoferm®, and 0.12%v/v Neutrase® at 782 rpm stirring speed and 60 min extraction time. The optimum extract yielded TPC of 122.1 ± 3.58 mg GAE/g DW, TFC of 47.3 ± 2.0 mg CAE/g DW, and antioxidant activity with an IC₅₀ of 0.72 ± 0.07 µL extract /mL. A control experiment was carried out under identical conditions without enzyme addition and achieved TPC of 41.6 ± 1.8 mg GAE/g DW and TFC of 20.9 ± 1.2 mg CAE/g DW. The biological activities of the extracts revealed strong inhibition of E. coli growth, moderate tyrosinase inhibition, and weak α-amylase inhibition. Sustainability of the extraction method was confirmed through AGREEprep, a green analysis tool, in accordance with green analytical chemistry principles. This study highlights a fast and efficient multi-enzyme approach for extracting bioactive compounds from Rosa canina L. pseudofruit, yielding extracts with potential applicability across diverse industrial sectors, such as in cosmetology and in nutraceuticals.
Lemoni et al. (Sun,) studied this question.