Immobilization of Lipases on Heterofunctional Octyl-Glyoxyl Agarose Supports: Improved Stability and Prevention of the Enzyme Desorption.
Lipases are among the most widely used enzymes in industry. Here, a novel method is described to rationally design the support matrix to retain the enzyme on the support matrix without leaching and also activate the enzyme for full activity retention. Lipases are interesting biocatalysts because they show the so-called interfacial activation, a mechanism of action that has been used to immobilize lipases on hydrophobic supports such as octyl-agarose. Thus, adsorption of lipases on hydrophobic surfaces is very useful for one step purification, immobilization, hyperactivation, and stabilization of most lipases. However, lipase molecules may be released from the support under certain conditions (high temperature, organic solvents), as there are no covalent links between the enzyme and the support matrix. A heterofunctional support has been proposed in this study to overcome this problem, such as the heterofunctional glyoxyl-octyl agarose beads. It couples the numerous advantages of the octyl-agarose support to covalent immobilization and creates the possibility of using the biocatalyst under any experimental conditions without risk of enzyme desorption and leaching. This modified support may be easily prepared from the commercially available octyl-agarose. Preparation of this useful support and enzyme immobilization on it via covalent linking is described here. The conditions are described to increase the possibility of achieving at least one covalent attachment between each enzyme molecule and the support matrix.
Departamento de Biocatálisis, Instituto de Catálisis-CSIC, Campus UAM-CSIC, Madrid, Spain; Escuela de Química, Grupo de investigación en Bioquímica y Microbiología (GIBIM), Edificio Camilo Torres 210, Universidad Industrial de Santander, Bucaramanga, Colombia.
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Rueda
Departamento de Biocatálisis, Instituto de Catálisis-CSIC, Campus UAM-CSIC, Madrid, Spain; Departamento de Engenharia Química, Universidade Federal Do Ceará, Campus Do Pici, Fortaleza, Ceará, Brazil.
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Dos Santos
Escuela de Química, Grupo de investigación en Bioquímica y Microbiología (GIBIM), Edificio Camilo Torres 210, Universidad Industrial de Santander, Bucaramanga, Colombia.
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Torres
Escuela de Bacteriología y Laboratorio Clínico, Universidad Industrial de Santander, Bucaramanga, Colombia.
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Ortiz
Departamento de Química, Facultad de Ciencias, Universidad del Tolima, Ibagué, Colombia.
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Barbosa
Departamento de Biocatálisis, Instituto de Catálisis-CSIC, Campus UAM-CSIC, Madrid, Spain. Electronic address: rfl@icp.csic.es.
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Fernandez-Lafuente
D000327:Adsorption; D003432:Cross-Linking Reagents / Q000737:chemistry; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry*; D006038:Glyoxylates / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D008049:Lipase / Q000737:chemistry*; D010084:Oxidation-Reduction; D012685:Sepharose / Q000737:chemistry*; D013499:Surface Properties
D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D006038:Glyoxylates; C070399:glyoxyl agarose; D012685:Sepharose; D008049:Lipase
Glyoxyl; Immobilization; Interfacial-activation; Octyl; Purification; Stabilization