Strategies for Increasing Protein Stability

  • Peter G. Chandler
  • Sebastian S. Broendum
  • Blake T. Riley
  • Matthew A. Spence
  • Colin J. Jackson
  • Sheena McGowan
  • Ashley M. BuckleEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2073)


The stability of wild-type proteins is often a hurdle to their practical use in research, industry, and medicine. The route to engineering stability of a protein of interest lies largely with the available data. Where high-resolution structural data is available, rational design, based on fundamental principles of protein chemistry, can improve protein stability. Recent advances in computational biology and the use of nonnatural amino acids have also provided novel rational methods for improving protein stability. Likewise, the explosion of sequence and structural data available in public databases, in combination with improvements in freely available computational tools, has produced accessible phylogenetic approaches. Trawling modern sequence databases can identify the thermostable homologs of a target protein, and evolutionary data can be quickly generated using available phylogenetic tools. Grafting features from those thermostable homologs or ancestors provides stability improvement through a semi-rational approach. Further, molecular techniques such as directed evolution have shown great promise in delivering designer proteins. These strategies are well documented and newly accessible to the molecular biologist, allowing for rapid enhancements of protein stability.

Key words

Protein stability Rational design Consensus design Ancestral reconstruction Semi-rational design Directed evolution 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Peter G. Chandler
    • 1
  • Sebastian S. Broendum
    • 1
  • Blake T. Riley
    • 1
  • Matthew A. Spence
    • 1
  • Colin J. Jackson
    • 2
  • Sheena McGowan
    • 3
  • Ashley M. Buckle
    • 1
    Email author
  1. 1.Department of Biochemistry and Molecular Biology, Biomedicine Discovery InstituteMonash UniversityClaytonAustralia
  2. 2.Research School of ChemistryAustralian National UniversityCanberraAustralia
  3. 3.Department of Microbiology, Biomedicine Discovery InstituteMonash UniversityClaytonAustralia

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