The temperature profile of the reaction was investigated to
determine the optimum temperature by conducting reactions
with 40 mm3 of butyric acid, 142 mm3 of isoamyl alcohol
(1:3 molar ratio of acid and alcohol), and 20 mg of enzyme,
at various temperatures. The reaction mixture was incubated
at various temperatures for 12 h. The optimum temperature
for the synthesis in SCCO2 was 55 ◦C for Novozym 435,
50 ◦C for HPL and Lipolase 100T. In solvent free synthesis,
the optimum temperature was 50 ◦C for all the enzymes.