## 4 Discussion  
Conventional methods for extracting bioactive compounds from plants often involve the use of organic solvents, high temperatures, and long extraction times. While effective, these methods can have significant environmental impacts and high energy costs. Among conventional methods there are solvent extraction, ultrasound-assisted extraction, microwave-assisted extraction, and supercritical CO\({}_{2}\) extraction [40; 41; 42; 43]. On the other hand, enzymatic extraction represents a \"green\" method for extracting active compounds from plant materials. Figure 5: Ion abundance of luteolin, verbascoside and ferulic acid-hexoside-hammoside (**A**) and coumaoyl-acetylhammosyl-aucubin, forsythoside B, caffeoyl-acetylhammosyl-aucubin, caffeic and _p_-coumaric acid (**B**); caffeic acid derivative II, III, IV and V (**C**); caffeic acid derivative I, caffeic acid-hexoside-hammoside and ferulic acid derivative (**D**) in the _V. nigrum_ extracts. Ion abundance levels of components analyzed through LC-MS/MS were reported as mean values \(\pm\) SD. Analysis of variance (ANOVA) was carried out through [PERSON]’s HSD test. Different letters indicate significantly different values (_p_ < 0.05). The enzymatic approach utilizes specific enzymes to degrade plant cell walls allowing the release of the active compounds contained within the cells. The enzymes used in the present work include cellulases, xylanases, pectinases, and amylases, which may work synergistically. Indeed, enzymatic combinations can improve extraction yields of phenolics due to their wide spectrum of action, allowing the hydrolysis of different components of the cell wall and increasing the soluble phenolic fraction, and favoring the release of free phenolic acids [44]. Enzymatic extraction is considered environmentally friendly as it avoids the use of toxic solvents and reduces energy consumption, making the process safer for the environment and end-users. For these reasons, several researchers are currently working to optimize enzymatic-assisted extraction protocols from different plant sources [45; 46]. In the specific case of _Verbuscum_, conventional extraction methods often involve the use of hydroalcoholic solvents for prolonged periods of time to maximize the recovery of phenolic compounds and flavonoids [39; 47; 48]. However, these methods can be improved by integrating a preliminary enzymatic treatment. In this work we demonstrated the effective and increased extraction of (poly)phenols, flavonoids, and other phytocomplexes from _V. nigrum_ using a novel enzymatic treatment followed by traditional solvent extraction. The application of an enzymatic treatment followed by hydroalcoholic extraction accelerated the release of phenolic compounds, achieving maximum yields in only 2 h compared to the >24 h required for hydroalcoholic extraction without enzymatic treatment. These results confirmed the fact that enzymatic extraction improves the degradation of lignocellulosic, making active compounds more accessible and therefore easier to extract [26]. The two-step method takes advantage of enzymatic extraction to produce high-quality, bioactive extracts in a sustainable manner, breaking down the plant cell walls, enhancing the release of active compounds; in the second step, hydroalcoholic extraction isolates the target compounds. By combining these steps, the process maximizes the efficiency and yield of valuable compounds. Another benefit associated with the application of enzymatic pretreatment is related to the plethora of bioactives that can be obtained. Enzymes allow for the breakdown of complex plant matrices and release entrapped compounds more effectively. In our work, enzymatic extraction was shown to yield higher concentrations of some bioactive compounds such as \(p\)-coumaric acid, caffeic acid, and some other hydroxycinnamate derivatives. Although it affected the recovery of some compounds, it clearly increased the amount of other species, enriching the extracts into compounds that are usually not present in many industrial products.