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Desarrollo de biocatalizadores hidrofóbicos y termotolerantes mediante técnicas de evolución dirigida
Development of hydrofobic and thermotolerant biocatalysts by directed evolution techniques
Keywords:
Cloroperoxidasa, biocatalizadores, enzima, error-prone PCR, evolución dirigida (es)Cloroperoxidase, Biocatalysts, Enzyme, Error-prone PCR, Directed Evolution (en)
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La evolución dirigida se presenta hoy como una de las alternativas más efectivas en el desarrollo y adaptación de biocatalizadores a los requerimientos industriales. Este trabajo muestra el desarrollo de biocatalizadores más hidrofóbicos y termotolerantes útiles en procesos de biorefinación. Para la Industria del Petróleo. Se presenta la obtención de una segunda librería del gen de la enzima Cloroperoxidasa por Error-Prone PCR (EP-PCR) y la selección de los mejores mutantes de acuerdo con el grado de hidrofobicidad, termorresistencia y estabilidad en medios orgánicos. Estas proteínas fueron más activas que las obtenidas en la primera librería y que la proteína nativa. Se demuestra que esta técnica de evolución dirigida es eficiente en la generación de variedades enzimáticas de interés industrial.
Directed evolution has emerged as one of the most effective approaches in developing and adapting biocatalysts to industrial requirements. This work shows the development of hidrophobic and thermotolerant biocatalyst useful in biorefining processes for the Petroleum Industry. We generated a second library of Cloroperoxidase enzyme gene by Error-Prone PCR (EP-PCR) with the selection of the best mutants based on their hidrophobicity, thermoresistance and stability in organic media. These proteins were more active than the first library mutants and the wildtype enzyme. We demonstrated that this directed evolution technique is efficient in generating chimeric libraries of enzymes of industrial interest.
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Copyright (c) 2002 Revista Colombiana de Biotecnología
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