The effects of important reaction parameters for enhancing isoamyl acetate formation through lipase-catalyzed
esterification of isoamyl alcohol were investigated in this study. Increase in substrate (acid) concentration led to
decrease in conversions. A critical enzyme concentration of 3 g l
−1
was detected for a substrate concentration of 0.06
M (each of alcohol and acid). Solvents with partition coefficient higher than 1000 (log P3.0) supported enzyme
activity to give high conversions. Acetic acid at higher concentrations could not be esterified easily probably owing
to its role in lowering the microaqueous pH of the enzyme. Extraneous water/buffer addition decreased the isoamyl
acetate yields slightly (10%) at 0.005–0.01% v/v of the reaction mixture and drastically (40%) at above 0.01%
v/v. Buffer saturation of the organic solvent employed improved esterification (upto two-fold), particularly at
moderately higher substrate concentrations (0.18 M). Employing acetic anhydride instead of acetic acid resulted in
a two-fold increase in the yields (at 0.25 M substrate). Use of excess nucleophile (alcohol) concentration by increasing
the alcohol/acid molar ratio resulted in higher conversions in shorter duration (upto eight-fold even at 1.5 M acetic
acid). Yields above 80% were achieved with substrate concentrations as high as 1.5 M and more than 150 g l
−1
isoamyl acetate concentrations were obtained employing a relatively low enzyme concentration of 10 g l
−1
. The
operational stability of lipase was also observed to be reasonably high enabling ten reuses of the biocatalyst. © 2001
Elsevier Science B.V. All rights reserved.