title: Identification of a fungal 1,8-cineole synthase from Hypoxylon sp. with specificity determinants in common with the plant synthases.
abstract: Terpenes are an important and diverse class of secondary metabolites widely produced by fungi. Volatile compound screening of a fungal endophyte collection revealed a number of isolates in the family Xylariaceae, producing a series of terpene molecules, including 1,8-cineole. This compound is a commercially important component of eucalyptus oil used in pharmaceutical applications and has been explored as a potential biofuel additive. The genes that produce terpene molecules, such as 1,8-cineole, have been little explored in fungi, providing an opportunity to explore the biosynthetic origin of these compounds. Through genome sequencing of cineole-producing isolate E7406B, we were able to identify 11 new terpene synthase genes. Expressing a subset of these genes in Escherichia coli allowed identification of the hyp3 gene, responsible for 1,8-cineole biosynthesis, the first monoterpene synthase discovered in fungi. In a striking example of convergent evolution, mutational analysis of this terpene synthase revealed an active site asparagine critical for water capture and specificity during cineole synthesis, the same mechanism used in an unrelated plant homologue. These studies have provided insight into the evolutionary relationship of fungal terpene synthases to those in plants and bacteria and further established fungi as a relatively untapped source of this important and diverse class of compounds.
authors: From the Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520.
authors: Jeffrey J
authors: JJ
authors: Shaw
authors: From the Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520.
authors: Tetyana
authors: T
authors: Berbasova
authors: From the Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520.
authors: Tomoaki
authors: T
authors: Sasaki
authors: the Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, and.
authors: Kyra
authors: K
authors: Jefferson-George
authors: From the Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520.
authors: Daniel J
authors: DJ
authors: Spakowicz
authors: From the Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520.
authors: Brian F
authors: BF
authors: Dunican
authors: the Laboratorio de Biotecnología Vegetal, Pontificia Universidad Católica del Ecuador, Quito 17 01 21 84, Ecuador.
authors: Carolina E
authors: CE
authors: Portero
authors: the Laboratorio de Biotecnología Vegetal, Pontificia Universidad Católica del Ecuador, Quito 17 01 21 84, Ecuador.
authors: Alexandra
authors: A
authors: Narváez-Trujillo
authors: From the Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520, scott.strobel@yale.edu.
authors: Scott A
authors: SA
authors: Strobel
mesh_terms: D000595:Amino Acid Sequence; D001203:Ascomycota / Q000201:enzymology* / Q000378:metabolism; D019755:Carbon-Carbon Lyases / Q000737:chemistry* / Q000235:genetics; D003511:Cyclohexanols / Q000737:chemistry*; D060026:Endophytes / Q000201:enzymology; D000077591:Eucalyptol; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D039821:Monoterpenes / Q000737:chemistry*; D020125:Mutation, Missense; D010802:Phylogeny; D010940:Plant Proteins / Q000737:chemistry*; D018547:Plant Stems / Q000382:microbiology; D013379:Substrate Specificity; D055549:Volatile Organic Compounds / Q000378:metabolism
chemical_list: D003511:Cyclohexanols; D005656:Fungal Proteins; D039821:Monoterpenes; D010940:Plant Proteins; D055549:Volatile Organic Compounds; C019899:1,8-cineole synthetase; D019755:Carbon-Carbon Lyases; D000077591:Eucalyptol
keywords: Enzyme Mechanism; Fungi; Mutagenesis; Phylogenetics; Terpenoid
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