title: Relevance of substrates and products on the desorption of lipases physically adsorbed on hydrophobic supports.
abstract: Lipase B from Candida antarctica (CALB) has been physically immobilized on octyl-agarose via interfacial activation. The incubation of the enzyme in 80% ethanol at pH 5 and 25°C has not significant effect on enzyme activity. Moreover, the hydrolysis of 100mM tributyrin catalyzed by this biocatalyst exhibited a quite linear reaction course. However, a new cycle of tributyrin hydrolysis showed a drastic drop in the activity. SDS-PAGE gels of the supernatant and the biocatalyst showed a significant enzyme desorption after the reaction. Similar results could be appreciated using triacetin or sunflower oil, while using 300mM methyl phenyl acetate, butyl butyrate or ethyl butyrate most enzyme molecules remained immobilized. The results show that the detergent properties of some reaction products increase the enzyme release from the hydrophobic support, and this problem increased if the concentration of the reactants increased. Using 500mM tributyrin, even in fully aqueous medium, some enzyme desorption from the support may be observed. Thus, the results show a limitation of this kind of biocatalysts that should be considered in the selection of an industrial lipase biocatalyst.
authors.affiliation: Departamento de Biocatálisis, Instituto de Catálisis-CSIC, Campus UAM-CSIC, Madrid, Spain; Catedrático CONACYT - Centro de Investigación en Alimentación y Desarrollo, A.C., Centro de Innovación y Desarrollo Agroalimentario de Michoacán, A.C., Antigua Carretera a Pátzcuaro s/n, 58341, Morelia, Michoacán, Mexico.
authors.forename: Jose J
authors.initials: JJ
authors.lastname: Virgen-Ortíz
authors.affiliation: Departamento de Biocatálisis, Instituto de Catálisis-CSIC, Campus UAM-CSIC, Madrid, Spain; Unidad de Investigación y Desarrollo en Alimentos, Instituto Tecnológico de Veracruz, Calzada Miguel A. de Quevedo 2779, 91897, Veracruz, Mexico.
authors.forename: Veymar G
authors.initials: VG
authors.lastname: Tacias-Pascacio
authors.affiliation: Instituto de Química, Universidade Federal de Alfenas, 37130-000, Alfenas, MG, Brazil.
authors.forename: Daniela B
authors.initials: DB
authors.lastname: Hirata
authors.affiliation: Unidad de Investigación y Desarrollo en Alimentos, Instituto Tecnológico de Veracruz, Calzada Miguel A. de Quevedo 2779, 91897, Veracruz, Mexico.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Torrestiana-Sanchez
authors.affiliation: Instituto Tecnológico de Tuxtla Gutiérrez, Carretera Panamericana Km. 1080, Tuxtla Gutiérrez, Chiapas, Mexico.
authors.forename: Arnulfo
authors.initials: A
authors.lastname: Rosales-Quintero
authors.affiliation: Departamento de Biocatálisis, Instituto de Catálisis-CSIC, Campus UAM-CSIC, Madrid, Spain. Electronic address: rfl@icp.csic.es.
authors.forename: Roberto
authors.initials: R
authors.lastname: Fernandez-Lafuente
mesh_terms: D000327:Adsorption; D055162:Biocatalysis; D001709:Biotechnology; D002175:Candida / Q000201:enzymology; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D000431:Ethanol; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism*; D057927:Hydrophobic and Hydrophilic Interactions; D008049:Lipase / Q000737:chemistry / Q000378:metabolism*; D012685:Sepharose / Q000031:analogs & derivatives; D013379:Substrate Specificity; D014280:Triglycerides / Q000378:metabolism
chemical_list: D004800:Enzymes, Immobilized; D005656:Fungal Proteins; D014280:Triglycerides; D000431:Ethanol; C027176:octyl-sepharose CL-4B; D012685:Sepharose; D008049:Lipase; C431349:lipase B, Candida antarctica; C005830:tributyrin
keywords: Detergents; Enzyme leakage; Enzyme reuse; Lipase interfacial activation; Octyl agarose