Isotopic Labeling of Heterologous Proteins in the Yeast Pichia pastoris and Kluyveromyces lactis

  • Toshihiko Sugiki
  • Osamu Ichikawa
  • Mayumi Miyazawa-Onami
  • Ichio Shimada
  • Hideo Takahashi
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 831)

Abstract

Several protein expression systems are available for the preparation of stable isotope-labeled recombinant proteins for NMR studies. Yeast expression systems have several advantages over prokaryotic systems, such as the widely used Escherichia coli expression system. Protein expression using the methylotrophic yeast Pichia pastoris is commonly employed for the preparation of isotope-labeled proteins. Recently, the hemiascomycete yeast Kluyveromyces lactis expression system was reported as being useful for preparing proteins for NMR studies. Since each yeast expression system has different features, their applications have increased in number. In this chapter, we describe procedures for the efficient production of uniformly isotope-labeled proteins using the P. pastoris and the K. lactis yeast expression systems.

Key words

Yeast expression systems Pichia pastoris Kluyveromyces lactis Stable isotope labeling Fed-batch fermentation NMR 

Notes

Acknowledgments

This work was financially supported in part by the Ministry of Economy, Trade and Industry (METI) and the New Energy and Industrial Technology Development Organization (NEDO).

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Toshihiko Sugiki
    • 1
    • 2
  • Osamu Ichikawa
    • 3
  • Mayumi Miyazawa-Onami
    • 1
    • 2
  • Ichio Shimada
    • 2
    • 3
  • Hideo Takahashi
    • 2
    • 4
  1. 1.Japan Biological Informatics Consortium (JBiC)TokyoJapan
  2. 2.Biomedicinal Information Research Center (BIRC)National Institute of Advanced Industrial Science and Technology (AIST)TokyoJapan
  3. 3.Graduate School of Pharmaceutical SciencesThe University of TokyoTokyoJapan
  4. 4.Department of Supramolecular Biology, Graduate School of NanobioscienceYokohama City UniversityYokohamaJapan

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