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Evaluation of strategies to produce highly porous cross-linked aggregates of porcine pancreas lipase with magnetic properties

Autores:Roberto Fernandez-Lafuente
Grupos de investigación:Optimización de biocatalizadores y bioprocesos enzimáticos
José Renato Guimarães1, Raquel de Lima Camargo Giordano1, Roberto Fernandez-Lafuente2*, Paulo Waldir Tardioli1*
1   Graduate Program in Chemical Engineering, Department of Chemical Engineering, Federal University of São Carlos, Rod. Washington Luiz, São Carlos 13565-905, Brazil; renatoge74@gmail.com (J. R. G.); raquel@ufscar.br (R. L. C. G.); pwtardioli@ufscar.br (P.W.T).
2   Departmento de Biocatálisis, ICP-CSIC, Campus UAM-CSIC Madrid, Madrid 28049, Spain; rfl@icp.csic.es (R. F.-L.);

Abstract: The preparation of highly porous magnetic crosslinked aggregates (pm-CLEA) of porcine pancreas lipase (PPL) is here reported. Some strategies were evaluated to improve the volumetric activity of the immobilized biocatalyst, such as treatment of PPL with enzyme surface modifying agents (polyethyleneimine or dodecyl aldehyde), co-aggregation with protein co-feeders (bovine serum albumin and/or soy protein), use of silica magnetic nanoparticles functionalized with amino groups (SMNPs) as separation aid, and starch as pore-making agent. The combination of enzyme surface modification with dodecyl aldehyde, co-aggregation with SMNPs and soy protein, in the presence of 0.8% starch (followed by hydrolysis of the starch with α-amylase), yielded CLEAs expressing high activity (immobilization yield around 100% and recovered activity around 80%), high effectiveness factor (approximately 65% of the equivalent free enzyme activity) and high stability at 40 °C and pH 8.0, i.e., PPL CLEAs co-aggregated with SMNPs/bovine serum albumin or SMNPs/soy protein retained 80% and 50% activity after 10 h incubation, respectively, while free PPL was fully inactivated after 2 h. Besides, highly porous magnetic CLEAs co-aggregated with soy protein and magnetic nanoparticles (pm-SP-CLEAs) showed good performance and reusability in the hydrolysis of tributyrin for five 4h-batches.
Palabras clave:silica magnetic nanoparticles, bovine serum albumin, soy protein, starch, protein surface modifiers
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