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RevistaBiotechnology Progress

1,3-Regiospecific ethanolysis of soybean oil catalyzed by crosslinked porcine pancreas lipase aggregates

Autores:Roberto Fernandez-Lafuente
Grupos de investigación:Optimización de biocatalizadores y bioprocesos enzimáticos
Margarita D. Ramos1, Letícia P. Miranda1, Raquel L. C. Giordano1, Roberto Fernandez-
Lafuente2, William Kopp3*, Paulo W. Tardioli1*
1Departamento de Engenharia Química, PPG-EQ, Universidade Federal de São Carlos
(UFSCar), Rod. Washington Luiz, km 235, 13565-905 São Carlos, SP, Brazil.
2Departamento de Biocatálisis. ICP-CSIC, Campus UAM-CSIC Madrid, Spain. 
3Kopp Technologies (KTech), Rua Alfredo Lopes, 1717, Jardim Macarengo, 13560-460 São
Carlos, SP, Brazil. 
The preparation of crosslinked aggregates of pancreatic porcine lipase (PPL-CLEA) was
systematically studied, evaluating the influence of three precipitants and two crosslinking agents,
as well as the use of soy protein as an alternative feeder protein on the catalytic properties and
stability of the immobilized PPL. Standard CLEAs showed a global yield (CLEA’ observed
activity / offered total activity) of less than 4%, whereas with the addition of soy protein
(PPL:soy protein mass ratio of 1:3) the global yield was approximately 5-fold higher. The CLEA
of PPL prepared with soy protein as feeder (PPL:soy protein mass ratio of 1:3) and
glutaraldehyde as crosslinking reagent (10 μmoles of aldehyde groups/mg of total protein) was
more active mainly because of the reduced enzyme leaching in the washing step. This CLEA,
named PPL-SOY-CLEA, had an immobilization yield around 60% and an expressed activity
around 40%. In the ethanolysis of soybean oil, the PPL-SOY-CLEA yielded maximum fatty acid
ethyl ester (FAEE) concentration around 12-fold higher than that achieved using soluble PPL (34
h reaction at 30 oC, 300 rpm stirring, soybean oil/ethanol molar ratio of 1:5) with an enzyme load
around 2-fold lower (very likely due to free enzyme inactivation). The operational stability of the
PPL-SOY-CLEA in the ethanolysis of soybean oil in a vortex flow type reactor showed that
FAEE yield was higher than 50% during ten reaction cycles of 24 h. This reactor configuration
may be an attractive alternative to the conventional stirred reactors for biotransformations in
industrial plants using carrier-free biocatalysts.

Palabras clave:porcine pancreas lipase; soy protein; CLEAs; biodiesel; vortex flow reactor
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