Optimization of enzyme assisted reflux extraction process for extracting essential oil from the root of Litsea Cubeba by response surface methodology

Abstract

This study evaluated the effects of pectinase, cellulase, and hemicellulase on dried roots of Litsea cubeba powder using enzyme-assisted hydrodistillation. Through single-factor experiment, four factors, namely, enzyme dosage, liquid-to-material ratio, enzymatic hydrolysis time and distillation time, have significant influence on the final yield of essential oil. The response surface experiment was designed by Box-Behnken design principle. The obtained model F regression value = 79.48, P < 0.0001, indicating that the model is extremely significant. Combined with actual use scenarios, the optimal extraction conditions are enzyme dosage 1000 U/g, liquid-to-material ratio 15:1, enzymatic hydrolysis time 3 h, distillation time 4 h, and the yield of essential oil is 0.403%. In addition, essential oil has certain antibacterial activity against Fusarium graminearum, and the EC50 value of essential oil is 78.266 μg/mL. Analysis of dry weight, relative conductivity, MDA content, SOD, CAT and POD activity of hyphae. The antibacterial mechanism of essential oil is related to the destruction of hyphae cell membrane, causing membrane lipid peroxidation reaction, affecting protective enzyme activity, etc.