W.S.D. Silva et al. / Journal of Molecular Catalysis B: Enzymatic 68 (2011) 98–103
103
Finally, for all tested cases, enzymes and ILs could be recov-
ered and reused without any loss of activity. Enzyme was filtered,
washed with acetone and reused without any loss of activity. The
ILs were easily recovered after enzyme filtration and reused several
times after removal of solvent.
was employed: 30% A + 70% B in 0 min, 100% B in 15 min, 50%
B + 50% C in 30 min, followed by isocratic elution with 50% B + 50% C
for the last 20 min. Epoxidation reaction. In a thermo-controlled
bath (30 0.1 ◦C) enzyme lipase (100 mg), IL (1 mL), methyl oleate
(2 mL) and hydrogen peroxide (30%, v/v, 1 mL) were added to a
Schelenk flask. The mixture was stirred for five hours and the prod-
uct removed and directed analyzed by HPLC. Enzyme and IL reuse.
All enzymes could be efficiently recovered and reused. Enzyme can
be filtered, washed with acetone and reused without loss of activ-
ity. The ILs can be easily recovered after filtered the enzyme and
could be reused several times after solvent remove (vacuum).
3. Conclusions
Overall, the use of hydrophobic ILs (BMI.NTf2 and BMI.PF6) allow
the EMO formation in high yields and within a few hours and, in
many cases, in just one hour of reaction. It is preferred to carry out
the EMO formation under the tested conditions, with low yields of
DIOL. The use of a hydrophilic IL (BMI.BF4) has shown that DIOL
formation is viable and competes directly with the EMO formation.
However, the highest yield of EMO (89%) was obtained using the
enzyme from Aspergillus niger in this medium. The reaction takes
place in just one hour. Moreover, the DIOL yield can be increased
in hydrophilic ILs. This result is one of the best ever published
considering both selectivity and reaction time. Moreover, the use
of a hydrophobic IL favors the competition reaction of epoxida-
tion and hydrolysis, but epoxidation is much faster. And free acid
content is less than 10% after 6 h of reaction. It is also notewor-
thy that no other product could be detected during the reaction,
indicating the high selectivity of the catalytic system described
herein.
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General: Enzymes and all other chemicals were purchased from
further purification. We used standard and commonly used pro-
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of EMO, DIOL and methyl oleate were used to determine the HPLC
method [28]. HPLC analyses were performed on a Shimadzu LC-20A
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use and degassed by continuous stripping with nitrogen. Injection
volumes of 1 mL and a flow rate of 1 mL min−1 were used in all
analyses. All samples were dissolved in propan-2-ol-n-hexane (5:4,
v/v). All solvents were HPLC-grade and were used as purchased,
without further purification. A Shim-pack VP-ODS (particle size
4.60 mm, 250P 4.6 mm I.D.) column was obtained from Shimadzu.
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