The
Malaysian Journal of Analytical Sciences Vol 12 No 3 (2008): 575 - 585
BUTYL ACETATE SYNTHESIS USING IMMOBILIZED LIPASE IN
CALCIUM ALGINATE BEADS
Mohd Zulkhairi Abdul Rahim1, Pat M. Lee2* and Kong.H. Lee2
1Universiti Kuala Lumpur
— Malaysian Institute of Chemical and Bioengineering Technology (UniKL — MICET),
78000,
Alor Gajah, Melaka.
2Faculty of Applied Sciences, Universiti Teknologi
MARA,
40450
Shah Alam, Selangor.
*Corresponding author: drpatlee@salam.uitm.edu.my
Abstract
The esterification
reaction of acetic acid and n-butanol using immobilizd lipase encapsulated in calcium
alginate beads (Lipase — CAB) and in chitosan coated calcium alginate beads (Lipase-CCAB)
in n-hexane under mild reaction conditions were studied. Effects of temperature
and substrate concentration (acetic acid and n-butanol) using Lipase — CAB, Lipase
— CCAB and free lipase on the esterification reaction and their thermal stability towards esterification reaction were investigated. Results
of temperature studies showed that the butyl acetate conversion increased with increase
of temperature and reached the highest yield of about 70% around 50oC for both immobilized systems but the yield of product catalysed
by free enzyme decreased as temperature was
increased. Thermal stabilities studies showed
that the Lipase-CCAB and Lipase-CAB were stable throughout the temperature range
of 30-60oC, however, free lipase became less stable at temperatures higher
than 50oC. The substrates, n-
butanol and acetic acid exerted different effects on the esterification reaction and the reaction was favoured by higher
acetic acidconcentration than butanol. Kinetics parameters, Km and Vmax values for both substrates and the specific activities
of the three enzyme system were also determined. The beads morphology was examined
using SEM. Batch-wise operational stability studies for both
immobilized systems demonstrated that the immobilized lipase performed better in
the batchwise reactor system than the continuous
bioreactor system and that the immobilized lipase remained
active for at least
5 cycles
of batchwise esterification reactions.
Keywords: Enzyme, lipase, esterification, calcium alginate,
chitosan
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