Influence of reaction time The experiments of optimization
of the enzymatic reaction conditions were carried out with FFAs
from UVO (used vegetable oil) and the best conditions were applied
to FFAs extracted from the microalgal biomass. Novozym 435 was
chosen to catalyse this reaction because it is one of the most widely
used lipases in industry and for biodiesel production (7,14e17).
These experiments were firstly carried out at small scale (4 g of
FFAs from UVO), using a 1.5:1 methanol/FFA molar ratio and a 0.1
w/w lipase/FFA ratio (40C and 200 rpm). In these conditions it
was observed that for a reaction time of 0.5 h a 92.8% conversion
was attained, and this conversion kept practically constant until
24 h (around 94.6% at 2, 6 and 24 h). This reaction was scaled-up
maintaining the same operational conditions. Fig. 2 shows that at
0.5 h the conversion is again over 90% and the equilibrium yields
attained at both scales are similar (about 95%).
In the following experiment, the conditions previously applied
to FFAs from UVO were applied to FFAs extracted from the microalga
N. gaditana, although in this case 14.3 g of microalgal FFAs were
used (Fig. 2). It was observed that the esterification degree (ED)
exceeded 90% at 0.25 h of reaction, attaining values of over 95% at
only 0.5 h, i.e., the reaction velocity and conversions were equal to
or better than those obtained with FFAs from UVO.
These data (Fig. 2) show the high velocity of transformation of
FFAs to methyl esters, which is much faster than the transesterification
reaction velocity (acylglycerols þ alcohol). This reaction
velocity is higher than that obtained by Da Rós et al. (15) in
the production of biodiesel from cyanobacterial lipids, also using
Novozym 435; these authors attained an ethyl ester yield of 98.1%
after reaction times of 48 h using iso-octane as reaction media. This
result is logical because, in fact, the formation of methyl esters from
SLs by transesterification occurs in a two-step process, that is, the
oil is first hydrolysed to FFAs and partial glycerides and the FFAs
produced are then esterified with methanol (18). By the procedure
assayed in this work the first step of hydrolysis is avoided.
Influence of reaction time The experiments of optimizationof the enzymatic reaction conditions were carried out with FFAsfrom UVO (used vegetable oil) and the best conditions were appliedto FFAs extracted from the microalgal biomass. Novozym 435 waschosen to catalyse this reaction because it is one of the most widelyused lipases in industry and for biodiesel production (7,14e17).These experiments were firstly carried out at small scale (4 g ofFFAs from UVO), using a 1.5:1 methanol/FFA molar ratio and a 0.1w/w lipase/FFA ratio (40C and 200 rpm). In these conditions itwas observed that for a reaction time of 0.5 h a 92.8% conversionwas attained, and this conversion kept practically constant until24 h (around 94.6% at 2, 6 and 24 h). This reaction was scaled-upmaintaining the same operational conditions. Fig. 2 shows that at0.5 h the conversion is again over 90% and the equilibrium yieldsattained at both scales are similar (about 95%).In the following experiment, the conditions previously appliedto FFAs from UVO were applied to FFAs extracted from the microalgaN. gaditana, although in this case 14.3 g of microalgal FFAs wereused (Fig. 2). It was observed that the esterification degree (ED)exceeded 90% at 0.25 h of reaction, attaining values of over 95% atonly 0.5 h, i.e., the reaction velocity and conversions were equal toor better than those obtained with FFAs from UVO.These data (Fig. 2) show the high velocity of transformation ofFFAs to methyl esters, which is much faster than the transesterificationreaction velocity (acylglycerols þ alcohol). This reactionvelocity is higher than that obtained by Da Rós et al. (15) inthe production of biodiesel from cyanobacterial lipids, also usingNovozym 435; these authors attained an ethyl ester yield of 98.1%after reaction times of 48 h using iso-octane as reaction media. Thisresult is logical because, in fact, the formation of methyl esters fromSLs by transesterification occurs in a two-step process, that is, theoil is first hydrolysed to FFAs and partial glycerides and the FFAsproduced are then esterified with methanol (18). By the procedureassayed in this work the first step of hydrolysis is avoided.
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