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PublicationJournal of Molecular Catalysis B: Enzymatic
Year2015
Volume118
Pages43-51
International

Production and immobilization of Geotrichum candidum lipase via physical adsorption on eco-friendly support: Characterization of the catalytic properties in hydrolysis and esterification reactions

Authors:Roberto Fernandez-Lafuente
Groups of research:Optimización de biocatalizadores y bioprocesos enzimáticos
Elisa Z. Ramosa, Rodney H. Miotti Júniora, Patrícia F. de Castroa, Paulo. W. Tardiolib, Adriano A. Mendesa, Roberto Fernandéz-Lafuentec, Daniela B. Hirataa*
 
 
 
a Instituto de Química, Universidade Federal de Alfenas, 37130-000 Alfenas, MG, Brazil.
b Departamento de Engenharia Química, Universidade Federal de São Carlos, 13565-905, São Carlos, SP, Brazil.
c Departamento de Biocatálisis, Instituto de Catálisis-CSIC, Campus UAM-CSIC, Cantoblanco, 28049 Madrid, Spain.



The present study reports the production of an extracellular lipase from Geotrichum candidum by submerged fermentation using cottonseed oil as inductor agent and its immobilization on poly-hydroxybutyrate (PHB) particles via physical adsorption. The catalytic properties of the biocatalyst prepared were determined in aqueous (hydrolysis of olive oil emulsion) and organic (ethyl linoleate synthesis) media. In the enzyme production, maximum hydrolytic activity of 22.91 IU/mL at 30° C and pH 5.2 was reached after 48 h of cultivation. A single protein band with an apparent molecular mass of 65 kDa was detected by SDS-PAGE analysis. The biocatalyst prepared by offering 75 mL of crude enzymatic extract (without cells) per gram of support exhibited maximum hydrolytic activity of 404.4 ± 2.3 IU/g at 37°C and pH 7.0, with a recovered activity percentage of around 40% and an immobilization yield of 59%. The optimal pH and temperature for both soluble and immobilized enzyme in the hydrolysis reaction was 8.0 and around 37-40°C. The biocatalyst was more thermally stable than the crude enzymatic extract at 35°C in 8h (46.2% and 23.7%, respectively) and slightly more stable at 45°C in 40 min (47.5% and 35.2%, respectively). In the esterification reaction, around 70% ester conversion was reached after 2 h of reaction under experimental conditions previously optimized by Central Composite Rotatable Design (CCRD). The biocatalyst retained 93% of its initial esterification activity after 6 successive cycles of esterification reaction. This biocatalyst is a promising one to catalyze reactions in aqueous and organic media.


Keywords:Lipase production; Geotrichum candidum; Physical adsorption; Poly-hydroxybutyrate particles; Catalytic properties
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