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PublicationJournal of Applied Polymer Science

Pilot-Scale Development of Core-Shell Polymer Supports for the Immobilization of Recombinant Lipase B from Candida antarctica and their Application in the Production of Ethyl Esters from Residual Fatty Acids

Authors:Roberto Fernandez-Lafuente
Groups of research:Optimization of biocatalysts and bioprocesses
Eliane Pereira Cipolattia,d‡, Martina Costa Cerqueira Pintoa,b‡, Julia de Macedo Roberta,
Tabita Pizoni da Silvaa,d, Thamires da Costa Beraltoa,d, Jorge G. F. Santos Jr.b, Rui de
Paula Vieira de Castroa, Roberto Fernandez-Lafuentec, Evelin Andrade Manoela,d, José
Carlos Costa da Silva Pintob*, Denise Maria Guimarães Freirea*
aDepartamento de Bioquímica, Instituto de Química, Universidade Federal do Rio de
Janeiro, Rio de Janeiro, Brazil.
 bPrograma de Engenharia Química, COPPE, Universidade Federal do Rio de Janeiro,
Rio de Janeiro, Brazil.
cDepartment of Biocatalysis, ICP-CSIC, Campus UAM-CSIC, Cantoblanco, 28049,
Madrid, Spain.
dDepartamento de Biotecnologia Farmacêutica, Faculdade de Farmácia, Universidade
Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
A recombinant lipase B from 26 Candida antarctica (LipB) in Pichia pastoris
was synthesized through submerged fermentation using crude glycerin as substrate. The
immobilization of this enzyme on the core-shell polymeric supports is an effective
alternative for its application. The supports with distinct levels of hydrophobicity were
produced through combined suspension and emulsion polymerization in pilot scale.
Particles with distinct compositions were synthesized (PMMA/PMMA; PMMA-co-
DVB/PMMA-co-DVB and PS-co-DVB/PS-co-DVB) and employed on the
immobilization of the produced lipase (LipB) and the commercial enzyme (CalB). The
morphological properties (specific area, average pore diameter, specific volume of pores
and hydrophobicity level) and the influence of the polymerization conditions on the
 morphology of the supports were studied. The thermal stability of such biocatalysts was
also investigated in the presence of calcium cation (Ca+2), maintained 100% of the activity
after 3h at 50 ºC when the PMMA-co-DVB/PMMA-co-DVB was employed. The
synthesized enzyme and supports manufactured in pilot scale were employed successfully
for production of esters using residual fatty acids as substrates, adding value to these raw
materials and increasing the ranges of possible applications.
Keywords:lipase B from Candida antarctica; polymeric supports; core-shell supports; scale-up; combined suspension and emulsion polymerization
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