This research study aimed to optimize innovative conditions for enhancing yield and quality of essential oil isolated from Lavandula angustifolia (La) based on rapid and innovative techniques. Ultrasonic-microwave assisted preceded by enzymolysis treatment was used to isolate essential oil (EO) of La. Response Surface Methodology was employed for the same purpose. The yield % of La essential oil (La.EO) obtained from combined enzymes treatment (cellulase and hemicellulase) was higher than that obtained by a single enzyme treatment. Chemical composition analysis of La.EO isolated was carried out using GC-MS. Morphological structure of La tissues was observed using SEM. The anti-free radical activity was assayed using DPPH and ABTS+ methods The isolation conditions consisted of the concentration of enzymes (12.5mg) with a range of (5–20mg), sonication time (52.5min) with a range of (15–90min) and microwave power (275W) with a range of (500–50W). The uses of coupling ultrasonic-microwave could be lead to forming of the acoustic cavitation and rupture of plant tissue. This facilitates the flow of liquid phase into the glandular trichomes in plant cells and enhances the absorption from the matrix of plant. Thus, enhances the efficiency of essential oil isolation from aromatic plant based on cavitation phenomenon. The results of the current work found that La.EO which isolated by using ultrasonic-microwave techniques under optimal conditions could be used as food additives in food industries to prevent the oxidation and bacterial spoilage.