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Enzyme Evolution by Yeast Cell Surface Engineering

  • Natsuko Miura
  • Kouichi Kuroda
  • Mitsuyoshi Ueda
Part of the Methods in Molecular Biology book series (MIMB, volume 1319)

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.

Key words

Yeast cell surface engineering Protein evolution In vivo homologous recombination Firefly luciferase Substrate specificity and reactivity 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Natsuko Miura
    • 1
    • 2
  • Kouichi Kuroda
    • 1
  • Mitsuyoshi Ueda
    • 1
  1. 1.Division of Applied Life Sciences, Graduate School of AgricultureKyoto UniversitySakyo-ku, KyotoJapan
  2. 2.Radiation Oncology BranchNational Cancer Institute, National Institutes of HealthBethesdaUSA

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