Enzymes in Synthetic Biology

Volume 608 of the series Methods in Enzymology covers key aspects of enzyme discovery, engineering tools and platforms, and examples of applications in the enzymology of synthetic biology.

Section I: Enzyme Discovery 1. Enzyme Discovery: Enzyme Selection and Pathway Design P. Carbonell, Mathilde Koch, Thomas Duigou and J.L. Faulon

Section II: Enzyme Engineering Tools and Platforms 2. Cell-Free Synthetic Biology for Pathway Prototyping Ashty S. Karim and Michael C. Jewett 3. Fast and Flexible Synthesis of Combinatorial Libraries for Directed Evolution Joanna C. Sadler, Lucy Green, Neil Swainston, Douglas B. Kell and Andrew Currin

Section III: Enzyme Families: Parts and Platforms for Chemical Diversity 4. Sesquiterpene Synthase Catalyzed Conversion of a Farnesyl Diphosphate Analogue to a Non-natural Terpenoid Ether Florence Huynh David J. Miller and Rudolf K. Allemann 5. In Vivo Platforms for Terpenoid Overproduction and the Generation of Chemical Diversity Guangkai Bian, Tian Ma and Tiangang Liu 6. Imine Reductases, Reductive Aminases, and Amine Oxidases for the Synthesis of Chiral Amines: Discovery, Characterization, and Synthetic Applications S.C. Cosgrove, A. Brzezniak, S.P. France, J.I. Ramsden, J. Mangas-Sanchez, S.L. Montgomery, R.S. Heath and N.J. Turner 7. Experimental Protocols for Generating Focused Mutant Libraries and Screening for Thermostable Proteins M.J.L.J. Fürst, C. Martin, N. Loncar and M.W. Fraaije 8. Characterization of Cytochrome P450 Enzymes and their Applications in Synthetic Biology L. N. Jeffreys, H. M. Girvan, K. J. McLean and A. W. Munro

Section IV: Genome Engineering for Enzyme Pathways 9. Metabolic Engineering of Saccharomyces cerevisiae Using a Trifunctional CRISPR/Cas System for Simultaneous Gene Activation, Interference, and Deletion Carl Schultz, Jiazhang Lian and Huimin Zhao 10. YeastFab: High Throughput Genetic Parts Construction, Measurement and Pathway Engineering in Yeast Eva Garcia-Ruiz, Jamie Auxillos, Tianyi Li, Junbiao Dai and Yizhi Cai

Section V: Bioengineered Strains with Designed Enzyme Pathways 11. Halomonas and Pathway Engineering for Bioplastics Production Jiang Xiao-Ran, Yin Jin, Chen Xiangbin and Chen Guo-Qiang 12. An Engineering Approach for Rewiring Microbial Metabolism Sebastian Wenk, Oren Yishai, Steffen N. Lindner and Arren Bar-Even 13. Multifragment DNA Assembly of Biochemical Pathways via Automated Ligase Cycling Reaction C.J. Robinson, M.S. Dunstan, N. Swainston, J. Titchmarsh, E. Takano, N.S. Scrutton and A.J. Jervis 14. Commodity Chemicals From Engineered Multimodular Type I Polyketide Synthases Satoshi Yuzawa, Amin Zargar, Bo Pang, Leonard Katz and Jay D. Keasling 15. Modular Pathway Rewiring of Yeast for Amino Acid Production Quanli Liu, Tao Yu, Kate Campbell, Jens Nielsen and Yun Chen

• Detailed methods for laboratory use of enzymes in synthetic biology applications
• Informative case history examples illustrating how enzyme and metabolic engineering are used to generate new products
• Emphasises latest developments in laboratory automation for the engineering of biology
• Covers many aspects of the design, build, test, learn cycle used in synthetic biology