title: Preparation of a biocatalyst via physical adsorption of lipase from Thermomyces lanuginosus on hydrophobic support to catalyze biolubricant synthesis by esterification reaction in a solvent-free system.
abstract: Lipase from Thermomyces lanuginosus (TLL) was immobilized on mesoporous hydrophobic poly-methacrylate (PMA) particles via physical adsorption (interfacial activation of the enzyme on the support). The influence of initial protein loading (5-200mg/g of support) on the catalytic properties of the biocatalysts was determined in the hydrolysis of olive oil emulsion and synthesis of isoamyl oleate (biolubricant) by esterification reaction. Maximum adsorbed protein loading and hydrolytic activity were respectively ≈100mg/g and ≈650 IU/g using protein loading of 150mg/g of support. The adsorption process followed the Langmuir isotherm model (R(2)=0.9743). Maximum ester conversion around 85% was reached after 30min of reaction under continuous agitation (200rpm) using 2500mM of each reactant in a solvent-free system, 45°C, 20%m/v of the biocatalyst prepared using 100mg of protein/g of support. Apparent thermodynamic parameters of the esterification reaction were also determined. Under optimal experimental conditions, reusability tests of the biocatalyst (TLL-PMA) after thirty successive cycles of reaction were performed. TLL-PMA fully retained its initial activity up to twenty two cycles of reaction, followed by a slight decrease around 8.6%. The nature of the product (isoamyl oleate) was confirmed by attenuated total reflection Fourier transform infrared (ATR-FTIR), proton ((1)H NMR) and carbon ((13)C NMR) nuclear magnetic resonance spectroscopy analyses.
authors.affiliation: Institute of Chemistry, Federal University of Alfenas, 37130-000, Alfenas, MG, Brazil.
authors.forename: Flávia A P
authors.initials: FA
authors.lastname: Lage
authors.affiliation: Institute of Chemistry, Federal University of Alfenas, 37130-000, Alfenas, MG, Brazil.
authors.forename: Jaquelinne J
authors.initials: JJ
authors.lastname: Bassi
authors.affiliation: Institute of Chemistry, Federal University of Alfenas, 37130-000, Alfenas, MG, Brazil.
authors.forename: Maria C C
authors.initials: MC
authors.lastname: Corradini
authors.affiliation: Institute of Chemistry, Federal University of Alfenas, 37130-000, Alfenas, MG, Brazil.
authors.forename: Larissa M
authors.initials: LM
authors.lastname: Todero
authors.affiliation: Institute of Chemistry, Federal University of Alfenas, 37130-000, Alfenas, MG, Brazil.
authors.forename: Jaine H H
authors.initials: JH
authors.lastname: Luiz
authors.affiliation: Institute of Chemistry, Federal University of Alfenas, 37130-000, Alfenas, MG, Brazil. Electronic address: adriano_amendes@yahoo.com.br.
authors.forename: Adriano A
authors.initials: AA
authors.lastname: Mendes
mesh_terms: D000327:Adsorption; D001203:Ascomycota / Q000201:enzymology*; D055162:Biocatalysis; D004800:Enzymes, Immobilized / Q000378:metabolism; D004951:Esterification; D005656:Fungal Proteins / Q000378:metabolism*; D057927:Hydrophobic and Hydrophilic Interactions; D007700:Kinetics; D008049:Lipase / Q000378:metabolism*; D054327:Lubricants / Q000378:metabolism; D009829:Oleic Acids / Q000378:metabolism; D011109:Polymethacrylic Acids; D012997:Solvents
chemical_list: D004800:Enzymes, Immobilized; D005656:Fungal Proteins; D054327:Lubricants; D009829:Oleic Acids; D011109:Polymethacrylic Acids; D012997:Solvents; C030613:polymethacrylic acid; D008049:Lipase
keywords: Biolubricant synthesis; Interfacial activation; Lipase immobilization; Optimization; Poly-methacrylate particles; Spectroscopy analyses