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Selection of the Optimal Yeast Host for the Synthesis of Recombinant Enzymes

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Recombinant Protein Production in Yeast

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1923))

Abstract

Yeasts, like Arxula adeninivorans, Hansenula polymorpha, Pichia pastoris, Debaryomyces hansenii, Debaryomyces polymorphus, Schwanniomyces occidentalis, Yarrowia lipolytica, and Saccharomyces cerevisiae are frequently used producers of recombinant enzymes, particularly when posttranslational modifications are mandatory to obtain full functionality. The wide-range transformation/expression platform presented in this chapter can be used to select the optimal yeast host for high-level synthesis of the desired enzyme with favorable biochemical properties. This platform is composed of a selection marker and up to four expression modules in a linearized cassette. Here we describe the protocols for the assembly as well as the transformation of yeast strains with the respective cassettes, screening of transformants, the isolation and biochemical characterization of the enzymes, and finally a simple fermentation strategy to achieve maximal yields of the chosen recombinant enzyme.

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Acknowledgment

The work was supported by grants from the German Federal Ministry of Economics and Technology (Bundesministerium für Wirtschaft und Technologie, BMWi) within its program “Zentrales Innovationsprogramm Mittelstand, ZIM” (Grant Numbers KF2131620MD2, KF2131628SA4).

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Authors and Affiliations

  1. Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, Germany

    Felix Bischoff, Martin Giersberg, Falko Matthes, Tobias Schwalenberg, Sebastian Worch & Gotthard Kunze

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  1. Felix Bischoff
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  2. Martin Giersberg
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  3. Falko Matthes
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  4. Tobias Schwalenberg
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  5. Sebastian Worch
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  6. Gotthard Kunze
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Corresponding author

Correspondence to Gotthard Kunze .

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Editors and Affiliations

  1. Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) and Austrian Centre of Industrial Biotechnology (acib), Vienna, Austria

    Brigitte Gasser

  2. Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) and Austrian Centre of Industrial Biotechnology (acib), Vienna, Austria

    Diethard Mattanovich

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Bischoff, F., Giersberg, M., Matthes, F., Schwalenberg, T., Worch, S., Kunze, G. (2019). Selection of the Optimal Yeast Host for the Synthesis of Recombinant Enzymes. In: Gasser, B., Mattanovich, D. (eds) Recombinant Protein Production in Yeast. Methods in Molecular Biology, vol 1923. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9024-5_4

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  • DOI: https://doi.org/10.1007/978-1-4939-9024-5_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9023-8

  • Online ISBN: 978-1-4939-9024-5

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