title: Generation and Functional Evaluation of Designer Monoterpene Synthases.
abstract: Monoterpene synthases are highly versatile enzymes that catalyze the first committed step in the pathways toward terpenoids, the structurally most diverse class of plant natural products. Recent advancements in our understanding of the reaction mechanism have enabled engineering approaches to develop mutant monoterpene synthases that produce specific monoterpenes. In this chapter, we are describing protocols to introduce targeted mutations, express mutant enzyme catalysts in heterologous hosts, and assess their catalytic properties. Mutant monoterpene synthases have the potential to contribute significantly to synthetic biology efforts aimed at producing larger amounts of commercially attractive monoterpenes.
authors.affiliation: Institute of Biological Chemistry, M. J. Murdock Metabolomics Laboratory, Washington State University, Pullman, WA, United States.
authors.forename: N
authors.initials: N
authors.lastname: Srividya
authors.affiliation: Institute of Biological Chemistry, M. J. Murdock Metabolomics Laboratory, Washington State University, Pullman, WA, United States.
authors.forename: I
authors.initials: I
authors.lastname: Lange
authors.affiliation: Institute of Biological Chemistry, M. J. Murdock Metabolomics Laboratory, Washington State University, Pullman, WA, United States. Electronic address: lange-m@wsu.edu.
authors.forename: B M
authors.initials: BM
authors.lastname: Lange
mesh_terms: D001709:Biotechnology / Q000379:methods; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D019753:Intramolecular Lyases / Q000235:genetics* / Q000378:metabolism*; D031337:Mentha spicata / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D039821:Monoterpenes / Q000378:metabolism*; D016296:Mutagenesis; D009154:Mutation; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D039821:Monoterpenes; D010940:Plant Proteins; D011994:Recombinant Proteins; D019753:Intramolecular Lyases; C040265:pinene cyclase I
keywords: Isoprenoid; Mechanism; Mutagenesis; Protein engineering; Terpene synthase; Terpenoid