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Methods in Enzymology. ‘Rational Design of Enzyme-Nanomaterials’

Autores:José Cleiton Sousa dos Santos, Roberto Fernandez-Lafuente
Grupos de Investigación:Optimization of biocatalysts and bioprocesses
Lipases are among the most widely used enzymes. They are peculiar biocatalysts that suffer the so-called interfacial activation versus hydrophobic surfaces, a mechanism of action that has been used to immobilize lipases on hydrophobic supports such as octyl-agarose. This immobilization method is very useful and permits the 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). A heterofunctional support has been proposed to overcome this problem, the heterofunctional glyoxyl-octyl agarose beads. It couples the previous advantages of the octyl-agarose support to the possibility of using the biocatalyst under any experimental conditions without risk of enzyme desorption, and the support may be easily prepared from the commercial octyl-agarose. Here, we show how to prepare this useful support and how the enzymes should be immobilized on it to increase the possibility of achieving at least an enzyme-support covalent bond.
 Nazzoly Ruedaa,b, Jose C. S. dos Santosa,c, Rodrigo Torresb,d,
Claudia Ortize, Oveimar Barbosaf, Roberto Fernandez-Lafuentea*
a Departamento de Biocatálisis. Instituto de Catálisis-CSIC, Campus UAM-CSIC Madrid. Spain.
b Escuela de Química, Grupo de investigación en Bioquímica y Microbiología (GIBIM), Edificio Camilo Torres 210, Universidad Industrial de Santander, Bucaramanga, Colombia.
c Departamento de Engenharia Química, Universidade Federal Do Ceará, Campus Do Pici, CEP 60455-760, Fortaleza, CE, Brazil
d Current address: Laboratorio de Biotecnología, Instituto Colombiano del Petróleo-Ecopetrol, Piedecuesta, Bucaramanga, Colombia.
eEscuela de Bacteriología y Laboratorio Clínico, Universidad Industrial de Santander, Bucaramanga, Colombia.
f Departamento de Química, Facultad de Ciencias. Universidad del Tolima, Ibagué, Colombia
Key words: interfacial-activation, immobilization, stabilization, purification, octyl, glyoxyl.
Palabras clave:Immobilization, stabilization, interfacial-activation, purification, octyl, glyoxyl
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