Survey of Protein Engineering Strategies
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- Abstract
- Table of Contents
- Figures
- Literature Cited
Abstract
Protein engineering is altering the structure of a protein to improve or change its properties. This unit summarizes concepts for protein engineering using rational design, directed evolution, and combinations of them. Different strategies are presented for identifying the best mutagenesis method, how to identify desired variants by screening or selection, and examples for successful applications are given. This should enable researchers to choose the most promising tools to solve their protein engineering challenges. Curr. Protoc. Protein Sci. 66:26.7.1?26.7.14. © 2011 by John Wiley & Sons, Inc.
Keywords: protein engineering; rational protein design; directed evolution; biocatalysis
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- Introduction
- Current Strategies to Engineer Proteins
- Strategies to Engineer Specific Properties
- Mutagenesis Methods
- Conclusions
- Acknowledgements
- Literature Cited
- Figures
- Tables
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Figure 26.7.1 The native, folded conformation of protein (N) equilibrates with the denatured, unfolded conformation (D). The free energy of unfolding, ΔGD‐N , typically favors the folded conformation by 5‐15 kcal/mol, but this value changes under different conditions. The denatured conformation can irreversibly form insoluble aggregates. Abbreviations: aa, amino acid; E, enantioselectivity. View Image -
Figure 26.7.2 (A ) An engineered nitrilase catalyzes a key enantioselective step in synthesis of an atorvastatin (Lipitor) precursor under industrial scale conditions. (B ) Screening for nitrilase variants with increased enantioselectivity used mass spectrometry and 15 N‐labeled pseudo‐prostereogenic 3‐hydroxyglutaronitrile. View Image -
Figure 26.7.3 An engineered ( R )‐transaminase catalyzes the highly efficient enantioselective formation of a chiral amine for the synthesis of Sitagliptin. The process is superior to the chemical asymmetric reduction. View Image
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Literature Cited
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| Key References | |
| Lutz and Bornscheuer, 2009. See above. | |
| This two volume edited book comprehensively covers current protein engineering methods. |
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