title: Computational design and selections for an engineered, thermostable terpene synthase.
abstract: Terpenoids include structurally diverse antibiotics, flavorings, and fragrances. Engineering terpene synthases for control over the synthesis of such compounds represents a long sought goal. We report computational design, selections, and assays of a thermostable mutant of tobacco 5-epi-aristolochene synthase (TEAS) for the catalysis of carbocation cyclization reactions at elevated temperatures. Selection for thermostability included proteolytic digestion followed by capture of intact proteins. Unlike the wild-type enzyme, the mutant TEAS retains enzymatic activity at 65°C. The thermostable terpene synthase variant denatures above 80°C, approximately twice the temperature of the wild-type enzyme.
authors: Department of Chemistry, University of California, Irvine, California 92697-2025, USA.
authors: Juan E
authors: JE
authors: Diaz
authors: Chun-Shi
authors: CS
authors: Lin
authors: Kazuyoshi
authors: K
authors: Kunishiro
authors: Birte K
authors: BK
authors: Feld
authors: Sara K
authors: SK
authors: Avrantinis
authors: Jonathan
authors: J
authors: Bronson
authors: John
authors: J
authors: Greaves
authors: Jeffery G
authors: JG
authors: Saven
authors: Gregory A
authors: GA
authors: Weiss
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000737:chemistry* / Q000378:metabolism*; D019295:Computational Biology / Q000379:methods*; D018360:Crystallography, X-Ray; D055550:Protein Stability; D013696:Temperature; D013816:Thermodynamics; D014026:Nicotiana / Q000201:enzymology
chemical_list: D019883:Alkyl and Aryl Transferases; C078217:5-epi-aristolochene synthase; C108877:terpene synthase