Directed enzyme evolution: Rational thoughts for non-rational experiments

Trabajo presentado en el XXXVII Congreso de la Sociedad Española de Bioquímica y Biología Molecular (SEBBM), celebrado en Granada (España) del 09 al 12 de septiembre de 2014.Over the past 20 years, directed evolution has been seen to be the most reliable approach to protein engineering. This tool that has revolutionized the manner in which proteins are manipulated in the laboratory in order to improve their application in distinct industrial settings and to engineer complex metabolic pathways opening a research ground in synthetic biology. By mimicking the mutation, recombination and selection processes that occur naturally in evolution, in vitro evolution provides a means of directing the evolution of genes toward specifi c goals in a manner that may not occur in a natural environment. Here, we summarize some successful examples from our laboratory to tailor ligninolytic genes (laccases, peroxidases, peroxygenases) in the budding yeast Saccharomyces cerevisiae. This lecture will pay special attention to the use of the DNA recombination machinery of S. cerevisiae to accelerate artifi cial evolution, complementing the traditional in vitro methods to generate tailor-made enzymes.Peer Reviewe