The objective of the present study was to immobilize lipase and use it for transesterification reaction in a fluorous solvent. Polymeric substance tylose (Tyl) was chosen as immobilization support. Tylose immobilized lipase from Candida rugosa (CRL) was subjected to fluorous solvents treatment and was found most stable in perfluorooctane (PFO). The immobilized Tyl-CRL was characterized for protein loading, solvent stability, surface morphology, restoration of secondary structure, and thermal stability. Immobilized CRL was further applied for the synthesis of phenethyl propionate in a fluorous solvent. All reaction parameters were optimized using the response surface method (RSM). Optimized reaction parameters were substrate ratio 1:3, immobilized lipase quantity 30 mg, and temperature 45 °C. Prepared biocatalyst was evaluated for recyclability, and it was found that catalytic activity decreased by 77 % up to the fifth recycle run. The optimized synthesis protocol showed a wide substrate scope. The docking study showed very good interaction of substrates with lipase supporting the experimental findings. The developed methodology offers a safer, more efficient approach to synthesizing various esters used as nature-identical flavors.