Optimization of solvent-free enzymatic esterification in eutectic substrate reaction mixture

The Candida rugosa lipase catalyzed esterification of (-)-menthol and lauric acid (LA) was studied in a eutectic mixture formed by both substrates((-)-menthol:LA 3:1, mol/mol). No additional reaction solvent was necessary, since the (-)-menthol:LA deep eutectic solvent (DES) acts as combined reaction medium and substrate pool. Therefore, the esterification is conducted under solvent-free conditions. The thermodynamic water activity (a(w)) was identified as a key parameter affecting the esterification performance in the (-)-menthol:LA DES. A response surface methodology was applied to optimize the esterification conditions in terms a(w), amount of C. rugosa lipase (m(CRL)) and reaction temperature. Under the optimized reaction conditions (a(w) = 0.55; m(CRL) =60 mg; T =45 °C), a conversion of 95 ± 1% LA was achieved (one day), the final (-)-menthyl lauric acid ester concentration reached 1.36 ± 0.04 M (2.25 days). The experimental product formation rate agreed very well with the model prediction.