Terpene Synthases, Volume 700 in the Methods in Enzymology series, continues the legacy of this highly respected laboratory standard with its first dedicated collection on this important family of enzymes. Terpene synthases are a diverse set of enzymes that use exquisite mechanisms to form complex (poly)cyclic hydrocarbon skeletons. Chapters in this new volume include Structural analysis by X-ray crystallography and cryo-EM, Understanding mechanisms using stable isotopes, substrate analogs, or computational tools, Engineering fusion enzymes, Ancestral terpene cyclases, as well as the Sequence, structure, and function of non-canonical terpene synthases.
1. Cover image - p. i 2. Title page - p. ii 3. Table of Contents - p. iii 4. Series Page - p. iii 5. Copyright - p. iii 6. Contributors - p. iii 7. Preface - p. iii 8. Chapter One: Methods for the preparation and analysis of a bifunctional class II diterpene synthase, copalyl diphosphate synthase from Penicillium fellutanum - p. 2 9. Chapter Two: Deciphering magnesium binding site and structure-function insights in a class II sesquiterpene cyclase - p. 26 10. Chapter Three: Structural biology of terpene synthases - p. 60 11. Chapter Four: Methods for the preparation and analysis of the diterpene cyclase fusicoccadiene synthase - p. 90 12. Chapter Five: Translational fusion of terpene synthases for metabolic engineering: Lessons learned and practical considerations - p. 123 13. Chapter Six: Isotopic labelings for mechanistic studies - p. 164 14. Chapter Seven: Understanding mechanisms of terpene synthases using substrate analogs - p. 188 15. Chapter Eight: Production of non-natural terpenoids through chemoenzymatic synthesis using substrate analogs - p. 208 16. Chapter Nine: Docking carbocations into terpene synthase active sites using chemically meaningful constraints—The TerDockin approach - p. 232 17. Chapter Ten: Mechanistic docking in terpene synthases using EnzyDock - p. 266 18. Chapter Eleven: Bioinformatic analysis of microbial type terpene synthase genes in plants - p. 294 19. Chapter Twelve: Ancestral terpene cyclases: From fundamental science to applications in biosynthesis - p. 312 20. Chapter Thirteen: Methods for the discovery and characterization of octocoral terpene cyclases - p. 344 21. Chapter Fourteen: Brewing coral terpenes—A yeast based approach to soft coral terpene cyclases - p. 374 22. Chapter Fifteen: Characterization of UbiA terpene synthases with a precursor overproduction system in Escherichia coli - p. 396 23. Chapter Sixteen: Functional analysis of transmembrane terpene cyclases involved in fungal meroterpenoid biosynthesis - p. 420 24. Chapter Seventeen: Vanadium haloperoxidases as noncanonical terpene synthases - p. 448 25. Chapter Eighteen: Identification and functional/structural analyses of large terpene synthases - p. 478
Terpene Synthases, Volume 700 in the Methods in Enzymology series, continues the legacy of this highly respected laboratory standard with its first dedicated collection on this important family of enzymes. Terpene synthases are a diverse set of enzymes that use exquisite mechanisms to form complex (poly)cyclic hydrocarbon skeletons. Chapters in this new volume include Structural analysis by X-ray crystallography and cryo-EM, Understanding mechanisms using stable isotopes, substrate analogs, or computational tools, Engineering fusion enzymes, Ancestral terpene cyclases, as well as the Sequence, structure, and function of non-canonical terpene synthases.
1. Cover image - p. i 2. Title page - p. ii 3. Table of Contents - p. iii 4. Series Page - p. iii 5. Copyright - p. iii 6. Contributors - p. iii 7. Preface - p. iii 8. Chapter One: Methods for the preparation and analysis of a bifunctional class II diterpene synthase, copalyl diphosphate synthase from Penicillium fellutanum - p. 2 9. Chapter Two: Deciphering magnesium binding site and structure-function insights in a class II sesquiterpene cyclase - p. 26 10. Chapter Three: Structural biology of terpene synthases - p. 60 11. Chapter Four: Methods for the preparation and analysis of the diterpene cyclase fusicoccadiene synthase - p. 90 12. Chapter Five: Translational fusion of terpene synthases for metabolic engineering: Lessons learned and practical considerations - p. 123 13. Chapter Six: Isotopic labelings for mechanistic studies - p. 164 14. Chapter Seven: Understanding mechanisms of terpene synthases using substrate analogs - p. 188 15. Chapter Eight: Production of non-natural terpenoids through chemoenzymatic synthesis using substrate analogs - p. 208 16. Chapter Nine: Docking carbocations into terpene synthase active sites using chemically meaningful constraints—The TerDockin approach - p. 232 17. Chapter Ten: Mechanistic docking in terpene synthases using EnzyDock - p. 266 18. Chapter Eleven: Bioinformatic analysis of microbial type terpene synthase genes in plants - p. 294 19. Chapter Twelve: Ancestral terpene cyclases: From fundamental science to applications in biosynthesis - p. 312 20. Chapter Thirteen: Methods for the discovery and characterization of octocoral terpene cyclases - p. 344 21. Chapter Fourteen: Brewing coral terpenes—A yeast based approach to soft coral terpene cyclases - p. 374 22. Chapter Fifteen: Characterization of UbiA terpene synthases with a precursor overproduction system in Escherichia coli - p. 396 23. Chapter Sixteen: Functional analysis of transmembrane terpene cyclases involved in fungal meroterpenoid biosynthesis - p. 420 24. Chapter Seventeen: Vanadium haloperoxidases as noncanonical terpene synthases - p. 448 25. Chapter Eighteen: Identification and functional/structural analyses of large terpene synthases - p. 478
