Enzyme Engineering and Evolution: Specific Enzyme Applications
- 1st Edition, Volume 644 - September 15, 2020
- Imprint: Academic Press
- Editor: Dan S. Tawfik
- Language: English
- Hardback ISBN:9 7 8 - 0 - 1 2 - 8 2 4 4 3 1 - 9
- eBook ISBN:9 7 8 - 0 - 1 2 - 8 2 4 4 3 2 - 6
Methods in Enzymology, Volume 644, the latest release in this ongoing serial, continues the legacy of this premier serial with quality chapters authored by leaders in the field.… Read more
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Request a sales quoteMethods in Enzymology, Volume 644, the latest release in this ongoing serial, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Chapters in this new release include Site-directed recombination (SDR) in vivo: a fast and reliable tool to unveil beneficial epistasis, Creation and application of amine oxidase with expanded substrate specificities from porcine kidney D-amino acid oxidase, Methods to assess correlation networks for engineering transketolase, Exploration of Enzyme Diversity by Integrating Bioinformatics with Microfluidics, Engineering lytic polysaccharide monooxygenases (LPMOs), Emulsion-based directed evolution of enzymes in yeast, and much more.
- Provides the authority and expertise of leading contributors from an international board of authors
- Presents the latest release in the Methods in Enzymology series
Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists
- Cover image
- Title page
- Table of Contents
- Copyright
- Contributors
- Preface
- Acknowledgments
- Chapter One: Engineering lytic polysaccharide monooxygenases (LPMOs)
- Abstract
- 1: Introduction
- 2: Engineering LPMO properties: Challenges and possible pitfalls
- 3: Engineering LPMOs: Optimization targets and how to possibly get there
- 4: Concluding remarks and future perspectives
- Acknowledgments
- Chapter Two: Strategies for the expression and characterization of artificial myoglobin-based carbene transferases
- Abstract
- 1: Introduction
- 2: Design of cofactor reconfigured myoglobin-based carbene transferases
- 3: Synthesis of iron porphyrinoid complexes
- 4: Incorporation of non-native cofactors into myoglobin
- 5: Catalytic activity
- 6: Summary
- Acknowledgments
- Chapter Three: Methods for the detection and analysis of dioxygenase catalyzed dihydroxylation in mutant derived libraries
- Abstract
- 1: Introduction
- 2: Heterologous expression of dioxygenase-type enzymes in E. coli
- 3: Indigo assay; a suitable colorimetric method for high-throughput screening of dioxygenase derived libraries
- 4: Analytical methods; GC and HPLC based strategies for the measurement and characterization of dioxygenase activity
- 5: Oxygen electrode; dioxygenase activity detection on real-time fashion via oxygen consumption
- 6: Summary
- Chapter Four: Enhancing promiscuous chemistries of a Schiff-base forming enzyme by divergent evolution
- Abstract
- 1: Introduction
- 2: Preparation of crude lysates for screening
- 3: Screening for improved retroaldol cleavage by UV/vis spectroscopy
- 4: HPLC-based screening for alternative aldol donor specificities
- 5: Screening for stereoselective Michael additions by chiral HPLC
- 6: Enhancing screening throughput for Knoevenagel activity using a colorimetric whole-cell assay
- 7: Summary & Conclusions
- Acknowledgments
- Chapter Five: Glutamine-walking: Creating reactive substrates for transglutaminase-mediated protein labeling
- Abstract
- 1: Introduction
- 2: Materials, equipment, reagents and services
- 3: Protocols
- 4: Analysis
- 5: Alternative methods/procedures
- 6: Troubleshooting and optimization
- 7: Summary
- Acknowledgments
- Chapter Six: Engineering aldolases for asymmetric synthesis
- Abstract
- 1: Introduction
- 2: Methods and results
- 3: Discussion
- 4: Conclusions
- Chapter Seven: Ultra-high throughput screening for novel protease specificities
- Abstract
- 1: Introduction
- 2: Materials, equipment and reagents
- 3: Protocols
- 4: Troubleshooting and optimization
- 5: Summary
- Acknowledgments
- Chapter Eight: Directed evolution of transcriptional switches using dual-selector systems
- Abstract
- 1: Introduction
- 2: Materials
- 3: Methods
- 4: Notes
- Chapter Nine: Methods for competitive enrichment and evaluation of superior DNA ligases
- Abstract
- 1: Introduction
- 2: General equipment
- 3: Competitive ligase enrichment
- 4: Fluorescence-based gel assay to quantify relative ligase activities
- Acknowledgments
- Chapter Ten: Directed evolution of custom polymerases using droplet microfluidics
- Abstract
- 1: Introduction
- 2: Library assembly
- 3: Droplet Sorting
- 4: Polymerase characterization
- 5: Notes
- Chapter Eleven: Optically guided mass spectrometry to screen microbial colonies for directed enzyme evolution
- Abstract
- 1: Introduction
- 2: Applications of optically guided MALDI-ToF MS to enzyme engineering
- 3: Conclusion
- Acknowledgments
- Chapter Twelve: Split enzymes: Design principles and strategy
- Abstract
- 1: Introduction to split enzymes
- 2: Ideal properties of a split enzyme system
- 3: Design criteria of a split enzyme system
- 4: Examples of split enzyme designs
- 5: Future outlook on split enzyme designs
- Acknowledgments
- Edition: 1
- Volume: 644
- Published: September 15, 2020
- Imprint: Academic Press
- No. of pages: 312
- Language: English
- Hardback ISBN: 9780128244319
- eBook ISBN: 9780128244326
DT
Dan S. Tawfik
Dan Tawfik is Principal Investigator at The Department of Biomolecular Sciences, Weizmann Institute of Science, Israel.
Affiliations and expertise
Principal Investigator, Department of Biomolecular Sciences, Weizmann Institute of Science, IsraelRead Enzyme Engineering and Evolution: Specific Enzyme Applications on ScienceDirect