Description
Book SynopsisThis 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.
Table of Contents1. 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
