Protein Engineering: Methods and Protocols

Front Matter ....Pages i-xi
 Protein Engineering: Past, Present, and Future  (Stefan Lutz, Samantha M. Iamurri)....Pages 1-12
Front Matter ....Pages 13-13
 Rational and Semirational Protein Design  (Ivan V. Korendovych)....Pages 15-23
 Computational Analysis of Protein Tunnels and Channels  (Jan Brezovsky, Barbora Kozlikova, Jiri Damborsky)....Pages 25-42
 YASARA: A Tool to Obtain Structural Guidance in Biocatalytic Investigations  (Henrik Land, Maria Svedendahl Humble)....Pages 43-67
 A Computational Library Design Protocol for Rapid Improvement of Protein Stability: FRESCO  (Hein J. Wijma, Maximilian J. L. J. Fürst, Dick B. Janssen)....Pages 69-85
 Directed Evolution of Proteins Based on Mutational Scanning  (Carlos G. Acevedo-Rocha, Matteo Ferla, Manfred T. Reetz)....Pages 87-128
Front Matter ....Pages 129-129
 A Brief Guide to the High-Throughput Expression of Directed Evolution Libraries  (Ana Luísa Ribeiro, Mario Mencía, Aurelio Hidalgo)....Pages 131-143
 Library Growth and Protein Expression: Optimal and Reproducible Microtiter Plate Expression of Recombinant Enzymes in E. coli Using MTP Shakers  (Sandy Schmidt, Mark Dörr, Uwe T. Bornscheuer)....Pages 145-156
 Normalized Screening of Protein Engineering Libraries by Split-GFP Crude Cell Extract Quantification  (Javier Santos-Aberturas, Mark Dörr, Uwe T. Bornscheuer)....Pages 157-170
 Functional Analysis of Membrane Proteins Produced by Cell-Free Translation  (Srujan Kumar Dondapati, Doreen A. Wüstenhagen, Stefan Kubick)....Pages 171-185
Front Matter ....Pages 187-187
 Practical Considerations Regarding the Choice of the Best High-Throughput Assay  (Carolin Mügge, Robert Kourist)....Pages 189-208
 High-Throughput Screening Assays for Lipolytic Enzymes  (Alexander Fulton, Marc R. Hayes, Ulrich Schwaneberg, Jörg Pietruszka, Karl-Erich Jaeger)....Pages 209-231
 Continuous High-Throughput Colorimetric Assays for α-Transaminases  (Egon Heuson, Jean-Louis Petit, Franck Charmantray, Véronique de Bérardinis, Thierry Gefflaut)....Pages 233-245
 Colorimetric High-Throughput Screening Assays for the Directed Evolution of Fungal Laccases  (Isabel Pardo, Susana Camarero)....Pages 247-254
 Directed Coevolution of Two Cellulosic Enzymes in Escherichia coli Based on Their Synergistic Reactions  (Min Liu, Lidan Ye, Hongwei Yu)....Pages 255-267
 Program-Guided Design of High-Throughput Enzyme Screening Experiments and Automated Data Analysis/Evaluation  (Mark Dörr, Uwe T. Bornscheuer)....Pages 269-282
 Solid-Phase Agar Plate Assay for Screening Amine Transaminases  (Martin S. Weiß, Uwe T. Bornscheuer, Matthias Höhne)....Pages 283-296
 Ultrahigh-Throughput Screening of Single-Cell Lysates for Directed Evolution and Functional Metagenomics  (Fabrice Gielen, Pierre-Yves Colin, Philip Mair, Florian Hollfelder)....Pages 297-309
 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 et al.)....Pages 311-331
 Library Generation and Auxotrophic Selection Assays in Escherichia coli and Thermus thermophilus  (Jörg Claren, Thomas Schwab, Reinhard Sterner)....Pages 333-345
Back Matter ....Pages 347-350