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RevistaRSC Advances
Año2016
Volumen6
Páginas4043-4052
Internacional

Evaluation of the performance of differently immobilized recombinant lipase B from Candida antarctica preparations for the synthesis of pharmacological derivatives in organic media

Autores:Roberto Fernandez-Lafuente
Grupos de investigación:Optimización de biocatalizadores y bioprocesos enzimáticos
Evelin A. Manoela,b*, Julia  M. Robertb, Martina C. C. Pintof, Antonio C. O. Machadob, Marina D. Bestetif, Maria Alice Z. Coelhod, Alessandro B. C. Simasc, Roberto Fernandez-Lafuentee*, Jose Carlos Pintof and Denise M. G. Freireb
a Laboratório Integrado de Pesquisas em Biotecnologia, Departamento de Biotecnologia Farmacêutica, Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
b Laboratório de Biotecnologia Microbiana, Departamento de Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
c Laboratório Roderick Barnes, Instituto de Pesquisas e Produtos Naturais, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.

d Biological System Engineering Group Laboratory, Departamento de Engenharia Bioquímica, Escola de Química, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.

 e Department of Biocatalysis, ICP-CSIC, Campus UAM-CSIC, Cantoblanco, 28049, Madrid, Spain
f Laboratório de Engenharia de Polímeros/EngePol, Programa de Engenharia Química, COPPE, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil. 


Abstract:
This paper shows the production of the lipase B from Candida antarctica (LIPB) after cloning the gene that encoded it in Pichia pastoris using PGK as a constitutive promoter.  The production of the lipase is lower using this strategy but it avoids the use of inducers like methanol. The performance of this enzyme was compared with that of the commercial enzyme (CALB) after immobilization on different supports in different reactions. As supports, we used Accurel 1000, and three core-shell supports (poly(methyl methacrylate) on the core and on the shell - PMMA/PMMA; poly(methyl methacrylate-co-divinylbenzene) on the core and on the shell - PMMA-co-DVB/PMMA-co-DVB; and poly(styrene-co-divinylbenzene) on the core and on the shell - PS-co-DVB/PS-co-DVB). The popular Novozym 435 was also utilized to assess the features of the new biocatalysts. All these supports adsorbed lipases via interfacial activation of the open form of the lipase on the hydrophobic surface of the supports. The studied reactions were esterification of oleic acid and ethanol in a solvent-free medium, resolution of (±)-1,3,5-O-benzyl-myo-inositol via acylation using vinyl acetate in hexane and resolution of  (±)-1,2-O-isopropylidene-3,6-di-O-benzyl-myo-inositol via acylation using vinyl acetate (solvent free system). Results varied depending on the employed supports and on the studied reactions, but some general trends may be observed, pointing to a better behavior of LIPB compared to CALB. The use of 4 different supports gave more strength to these differences, as it did not depend on a specific difference between a single support/enzyme pair, but it is more general.        Thus, LIPB seems to have some advantages compared to the commercial enzyme on all the reaction assayed in this paper. PS-co-DVB/PS-co-DVB-LIPB is in general the most active preparation (even 50% higher activity was observed). Further investigations are in development to determine the structural reasons for these differences.
Palabras clave:recombinant lipase B from Candida antarctica; core-shell supports; lipase immobilization; lipase interfacial activation; lipase improved properties.
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