An enzyme-assisted modified steam distillation process was adopted to extract turmeric oil from Curcuma longa L. rhizomes. The diastase, xylose, cellulase, pectinase, and lipase enzymes were used for the pre-treatment of fresh turmeric rhizome to obtain a higher yield by rapturing biological cells. The quantitative and qualitative analysis of turmeric oil was performed by GC–MS. The various influencing parameters for the extraction of turmeric oil such as an enzyme, incubation/pre-treatment time, distillation time have been studied in the presentwork. The obtained turmeric oil by enzymatic pretreatment process is richer in bioactive/medicinal components than in the other traditional methods. The maximum yield was obtained with cellulase enzyme, which is 25–27% higher than the yield obtained by the traditional hydro distillation process. The detailed qualitative and quantitative analyses are also presented. The present method can be considered energy-efficient, effective, economical, and eco-friendly.