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Insoluble Proteins pp 45–63Cite as

Optimization of Culture Parameters and Novel Strategies to Improve Protein Solubility

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1258))

Abstract

The production of recombinant proteins, in soluble form in a prokaryotic expression system, still remains a challenge for the biotechnologist. Innovative strategies have been developed to improve protein solubility in various protein overexpressing hosts. In this chapter, we would focus on methods currently available and amenable to “desired modifications,” such as (a) the use of molecular chaperones; (b) the optimization of culture conditions; (c) the reengineering of a variety of host strains and vectors with affinity tags; and (d) optimal promoter strengths. All these parameters are evaluated with the primary objective of increasing the solubilization of recombinant protein(s) during overexpression in Escherichia coli.

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Acknowledgments

We thank Ms. Sonam Grover, Ms. Reema Singh, and Mr. Abdullah Sheikh for the literature assistance and helpful comments. We thank Mr. Ravi Bharadwaj and Ms. Pratibha Chanana for optimizing conditions for GNE expression in E. coli at low temperature and collecting the data for Fig. 1. We are grateful to the Deanship of Scientific Research, King Abdulaziz University, Jeddah, Saudi Arabia, for the financial assistance.

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Authors and Affiliations

  1. School of Biotechnology, Jawaharlal Nehru University, New Delhi, 110067, India

    Ranjana Arya

  2. Department of Biology, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah, 21589, Saudi Arabia

    Jamal S. M. Sabir, Roop S. Bora & Kulvinder S. Saini

Authors
  1. Ranjana Arya
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  2. Jamal S. M. Sabir
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  3. Roop S. Bora
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  4. Kulvinder S. Saini
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Corresponding author

Correspondence to Kulvinder S. Saini .

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Editors and Affiliations

  1. Departament de Genètica i de Microbiologia, CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN) and Universitat Autònoma de Barcelona, Bellaterra (Barcelona), Spain

    Elena García-Fruitós

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Arya, R., Sabir, J.S.M., Bora, R.S., Saini, K.S. (2015). Optimization of Culture Parameters and Novel Strategies to Improve Protein Solubility. In: García-Fruitós, E. (eds) Insoluble Proteins. Methods in Molecular Biology, vol 1258. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2205-5_3

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  • DOI: https://doi.org/10.1007/978-1-4939-2205-5_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2204-8

  • Online ISBN: 978-1-4939-2205-5

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