title: Enzyme Evolution by Yeast Cell Surface Engineering.
abstract: Artificial evolution of proteins with the aim of acquiring novel or improved functionality is important for practical applications of the proteins. We have developed yeast cell surface engineering methods (or arming technology) for evolving enzymes. Here, we have described yeast cell surface engineering coupled with in vivo homologous recombination and library screening as a method for the artificial evolution of enzymes such as firefly luciferases. Using this method, novel luciferases with improved substrate specificity and substrate reactivity were engineered.
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan.
authors.forename: Natsuko
authors.initials: N
authors.lastname: Miura
authors.forename: Kouichi
authors.initials: K
authors.lastname: Kuroda
authors.forename: Mitsuyoshi
authors.initials: M
authors.lastname: Ueda
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D061505:Cell Surface Display Techniques / Q000379:methods; D020650:Combinatorial Chemistry Techniques; D048888:Fireflies / Q000201:enzymology; D059765:Homologous Recombination; D049409:Luciferases, Firefly / Q000096:biosynthesis* / Q000235:genetics*; D016297:Mutagenesis, Site-Directed; D019151:Peptide Library; D015202:Protein Engineering / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D019151:Peptide Library; D000255:Adenosine Triphosphate; D049409:Luciferases, Firefly