Enhancing the extraction yield and bioactivity of essential oil from Heracleum persicum seeds via enzyme pre-treatment

Heracleum persicum is an herbal plant traditionally used for treating various diseases. This study investigated the impact of enzyme pre-treatment before hydro-distillation (EP-HD) on the yield of essential oil (EO) extracted from Heracleum persicum seeds. Three independent variables, i.e., incubation temperature, incubation time, and enzyme concentration, were selected for optimization using the Box-Behnken design (BBD) of Response Surface Methodology (RSM). According to the RSM findings, the highest EO yield was achieved under the following conditions: an incubation temperature of 39.95 °C, an incubation time of 179 min, and an enzyme concentration of 0.445 % (w/v). The results confirmed that the EP-HD method significantly enhanced the EO yield, achieving 4.34 %, compared to conventional hydro-distillation (HD), which yielded 2.82 %. GC-FID analysis indicated that enzyme pre-treatment notably increased the percentages of hexyl butyrate and octyl acetate in the EO. Additionally, biological activity studies demonstrated that the EO obtained via EP-HD exhibited greater antioxidant and cytotoxic activities than EO produced by the HD method. Furthermore, the EO extracted using EP-HD displayed substantially stronger insecticidal properties (LD₅₀ = 24.13 μg/larva) compared to EO from the HD method (LD₅₀ = 33.19 μg/larva). These findings underscore the potential of enzyme pre-treatment as an effective technique for improving EO yield, reducing extraction time, and enhancing biological activities.