Heterologous Protein Expression and Secretion in Yarrowia lipolytica
Heterologous protein production constitutes an important research field, having both academic and commercial applications. The use of yeasts as host systems took advantage of the combination of their easy manipulation and high growth capacity, with their
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Abstract Heterologous protein production constitutes an important research field, having both academic and commercial applications. The use of yeasts as host systems took advantage of the combination of their easy manipulation and high growth capacity, with their eukaryotic subcellular organisation allowing posttranslational processing. Among available yeast host systems, Yarrowia lipolytica (Yarrowia) appears as one of the most attractive. This non-conventional dimorphic yeast has been distinguished due to its remarkable regularity of performance in the efficient secretion of various heterologous proteins. We will present in this chapter Yarrowia’s characteristics regarding heterologous protein production and describe the genetic and molecular tools available. Recent developments of the Yarrowia expression system, such as surface display vectors, engineered strains and highthroughput screening processes, will be reviewed. A comprehensive survey of the literature allowed us to list more than 100 heterologous proteins, from more than 60 species, successfully produced until now in this yeast. This amount of data sometimes offers a comparison with other yeast host systems and globally demonstrates the reliability and versatility of Yarrowia as host for heterologous production. Contents 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 1.1 Heterologous Expression in Yeast Host Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 1.2 Yarrowia as a Host for Heterologous Production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 1.3 Chronology of Heterologous Production in Yarrowia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 2 Genetic and Molecular Tools for Heterologous Production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 2.1 Expression/Secretion Vectors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 2.2 Recipient Strains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 2.3 Selection Marker Genes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 2.4 Expression Cassettes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 C. Madzak (*) • J.-M. Beckerich INRA, UMR1319 Micalis, Domaine de Vilvert, 78352 Jouy-en-Josas, France e-mail: [email protected]; [email protected] G. Barth (ed.), Yarrowia lipolytica, Microbiology Monographs 25, DOI 10.1007/978-3-642-38583-4_1, © Springer-Verlag Berlin Heidelberg 2013
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C. Madzak and J.-M. Beckerich
2.5 Development of a Commercial Yarrowia Expression Kit . . . . . . . . . . . . . . .
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