Protein Engineering: Methods and Protocols

ISBN-13: 9781493973644 / Angielski / Twarda / 2017 / 350 str.

Uwe T. Bornscheuer; Matthias Hohne

Opis

This volume details basic and advanced protocols for both stages of protein engineering: the library design phase and the identification of improved variants by screening and selection.

Spis treści

1. Protein Engineering - Past, Present, and Future

Stefan Lutz and Samantha M. Iamurri

 

Part I

2. (Semi)rational Protein Design

Ivan V. Korendovych

 

3. Computational Analysis of Protein Tunnels and Channels

Jan Brezovsky, Barbora Kozlikova, and Jiri Damborsky

 

4. YASARA - A Tool to Obtain Structural Guidance in Biocatalytic Investigations

Henrik Land and Maria Svedendahl Humble

 

5. A Computational Library Design Protocol for Rapid Improvement of Protein Stability - FRESCO

Hein J. Wijma, Maximilian J.L.J. Fürst, and Dick B. Janssen

 

6. Directed Evolution of Proteins Based on Mutational Scanning

Carlos G. Acevedo-Rocha, Matteo Ferla, and Manfred T. Reetz

 

Part II

7. A Brief Guide to the High-throughput Expression of Directed Evolution Libraries

Ana Luísa Ribeiro, Mario Mencía, and Aurelio Hidalgo

 

8. Library Growth and Protein Expression: Optimal and Reproducible Microtiter Plate Expression of Recombinant Enzymes in E. coli using MTP Shakers

Sandy Schmidt, Mark Dörr, and Uwe T. Bornscheuer

 

9. Normalized Screening of Protein Engineering Libraries by Split-GFP Crude Cell Extract Quantification

Javier Santos-Aberturas, Mark Dörr, and Uwe T. Bornscheuer

 

10. Functional Analysis of Membrane Proteins Produced by Cell-Free Translation

Srujan Kumar Dondapati, Doreen A Wüstenhagen, and Stefan Kubick

 

 

PART III

11. Practical Considerations Regarding the Choice of the Best High-throughput Assay

Carolin Mügge and Robert Kouristb

 

12. High-throughput Screening Assays for Lipolytic Enzymes

Alexander Fulton, Marc R. Hayes, Ulrich Schwaneberg, Jörg Pietruszka, and Karl-Erich Jaeger

 

13. Continuous High-throughput Colorimetric Assays for α-transaminases

Egon Heuson, Jean-Louis Petit, Franck Charmantray, Véronique de Bérardinis, and Thierry Gefflaut

 

14. Colorimetric High-troughput Screening Assays for the Directed Evolution of Fungal Laccases

Isabel Pardo and Susana Camarero

 

15. Directed Co-evolution of Two Cellulosic Enzymes in Escherichia coli Based on their Synergistic Reactions

Min Liu, Lidan Ye, and Hongwei Yu

 

16. Program-guided Design of High-throughput Enzyme Screening Experiments and Automated Data Analysis/Evaluation

Mark Dörr and Uwe T. Bornscheuer

 

17. Solid Phase Agar Plate Assay for Screening Amine Transaminases

Martin S. Weiß, Uwe T. Bornscheuer, and Matthias Höhne

 

18. Ultrahigh-throughput Screening of Single-cell Lysates for Directed Evolution and Functional Metagenomics

Fabrice Gielen, Pierre-Yves Colin, Philip Mair, and Florian Hollfelder

 

19. Isolation of pH-sensitive Antibody Fragments by Fluorescence-activated Cell Sorting and Yeast Surface Display

Christian Schröter, Simon Krah, Jan Beck, Doreen Könning, Julius Grzeschik, Bernhard Valldorf, Stefan Zielonka, and Harald Kolmar

 

20. Library Generation and Auxotrophic Selection Assays in Escherichia coli and Thermus thermophilus

Jörg Claren, Thomas Schwab, and Reinhard Sterner

Blurb

This volume details basic and advanced protocols for both stages of protein engineering: the library design phase and the identification of improved variants by screening and selection. Chapters focus on enzyme engineering using rational and semi-rational approaches. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Protein Engineering: Methods and Protocols aims to aid scientists in the planning and performance of their experiments.