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General Introduction: Recombinant Protein Production and Purification of Insoluble Proteins

  • Neus Ferrer-Miralles
  • Paolo Saccardo
  • José Luis Corchero
  • Zhikun Xu
  • Elena García-Fruitós
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1258)

Abstract

Proteins are synthesized in heterologous systems because of the impossibility to obtain satisfactory yields from natural sources. The production of soluble and functional recombinant proteins is among the main goals in the biotechnological field. In this context, it is important to point out that under stress conditions, protein folding machinery is saturated and this promotes protein misfolding and, consequently, protein aggregation. Thus, the selection of the optimal expression organism and the most appropriate growth conditions to minimize the formation of insoluble proteins should be done according to the protein characteristics and downstream requirements.

Escherichia coli is the most popular recombinant protein expression system despite the great development achieved so far by eukaryotic expression systems. Besides, other prokaryotic expression systems, such as lactic acid bacteria and psychrophilic bacteria, are gaining interest in this field. However, it is worth mentioning that prokaryotic expression system poses, in many cases, severe restrictions for a successful heterologous protein production. Thus, eukaryotic systems such as mammalian cells, insect cells, yeast, filamentous fungus, and microalgae are an interesting alternative for the production of these difficult-to-express proteins.

Key words

Recombinant proteins Protein expression Protein purification Aggregation Solubility Heterologous system Insoluble proteins 

Notes

Acknowledgments

The authors acknowledge the financial support granted to E.G.F. from Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria—MINECO (RTA2012-00028-C02-02) and Centro de Investigación Biomédica en Red (CIBER) de Bioingeniería, Biomateriales y Nanomedicina. Z.X. acknowledges financial support from China Scholarship Council. PS has received a predoctoral fellowship from Instituto de Salud Carlos III. The authors are also indebted to the Protein Production Platform (CIBER-BBN—UAB) for helpful technical assistance (http://www.bbn.ciber-bbn.es/programas/plataformas/equipamiento).

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Neus Ferrer-Miralles
    • 1
    • 2
  • Paolo Saccardo
    • 1
    • 2
  • José Luis Corchero
    • 1
    • 2
  • Zhikun Xu
    • 1
    • 2
  • Elena García-Fruitós
    • 3
    • 4
  1. 1.Departament de Genètica i de Microbiologia, Institut de Biotecnologia i de BiomedicinaUniversitat Autònoma de BarcelonaBarcelonaSpain
  2. 2.CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN)Universitat Autònoma de BarcelonaBarcelonaSpain
  3. 3.Departament de Genètica i de Microbiologia, Institut de Biotecnologia i de BiomedicinaUniversitat Autònoma de BarcelonaBarcelonaSpain
  4. 4.CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN)Universitat Autònoma de BarcelonaBarcelonaSpain

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