Structural Elucidation of Cisoid and Transoid Cyclization Pathways of a Sesquiterpene Synthase Using 2-Fluorofarnesyl Diphosphates

Sesquiterpene skeletal complexity in nature originates from the enzyme-catalyzed ionization of (trans,trans)-farnesyl diphosphate (FPP) (1a) and subsequent cyclization along either 2,3-transoid or 2,3-cisoid farnesyl cation pathways. Tobacco 5-epi-aristolochene synthase (TEAS), a transoid synthase, produces cisoid products as a component of its minor product spectrum. To investigate the cryptic cisoid cyclization pathway in TEAS, we employed (cis,trans)-FPP (1b) as an alternative substrate. Strikingly, TEAS was catalytically robust in the enzymatic conversion of (cis,trans)-FPP (1b) to exclusively (≥99.5%) cisoid products. Further, crystallographic characterization of wild-type TEAS and a catalytically promiscuous mutant (M4 TEAS) with 2-fluoro analogues of both all-trans FPP (1a) and (cis,trans)-FPP (1b) revealed binding modes consistent with preorganization of the farnesyl chain. These results provide a structural glimpse into both cisoid and transoid cyclization pathways efficiently templated by a single enzyme active site, consistent with the recently elucidated stereochemistry of the cisoid products. Further, computational studies using density functional theory calculations reveal concerted, highly asynchronous cyclization pathways leading to the major cisoid cyclization products. The implications of these discoveries for expanded sesquiterpene diversity in nature are discussed

Emergence of terpene cyclization in Artemisia annua

The emergence of terpene cyclization was critical to the evolutionary expansion of chemical diversity yet remains unexplored. Here we report the first discovery of an epistatic network of residues that controls the onset of terpene cyclization in Artemisia annua. We begin with amorpha-4,11-diene synthase (ADS) and (E)-b-farnesene synthase (BFS), a pair of terpene synthases that produce cyclic or linear terpenes, respectively. A library of B27,000 enzymes is generated by breeding combinations of natural amino-acid substitutions from the cyclic into the linear producer. We discover one dominant mutation is sufficient to activate cyclization, and together with two additional residues comprise a network of strongly epistatic interactions that activate, suppress or reactivate cyclization. Remarkably, this epistatic network of equivalent residues also controls cyclization in a BFS homologue from Citrus junos. Fitness landscape analysis of mutational trajectories provides quantitative insights into a major epoch in specialized metabolism

Variation in capsidiol sensitivity between Phytophthora infestans and Phytophthora capsici is consistent with their host range.

Plants protect themselves against a variety of invading pathogenic organisms via sophisticated defence mechanisms. These responses include deployment of specialized antimicrobial compounds, such as phytoalexins, that rapidly accumulate at pathogen infection sites. However, the extent to which these compounds contribute to species-level resistance and their spectrum of action remain poorly understood. Capsidiol, a defense related phytoalexin, is produced by several solanaceous plants including pepper and tobacco during microbial attack. Interestingly, capsidiol differentially affects growth and germination of the oomycete pathogens Phytophthora infestans and Phytophthora capsici, although the underlying molecular mechanisms remain unknown. In this study we revisited the differential effect of capsidiol on P. infestans and P. capsici, using highly pure capsidiol preparations obtained from yeast engineered to express the capsidiol biosynthetic pathway. Taking advantage of transgenic Phytophthora strains expressing fluorescent markers, we developed a fluorescence-based method to determine the differential effect of capsidiol on Phytophtora growth. Using these assays, we confirm major differences in capsidiol sensitivity between P. infestans and P. capsici and demonstrate that capsidiol alters the growth behaviour of both Phytophthora species. Finally, we report intraspecific variation within P. infestans isolates towards capsidiol tolerance pointing to an arms race between the plant and the pathogens in deployment of defence related phytoalexins

Oxidacion selectiva de metano a formaldehido sobre catalizadores vanadio-silice

Centro de Informacion y Documentacion Cientifica (CINDOC). C/Joaquin Costa, 22. 28002 Madrid. SPAIN / CINDOC - Centro de Informaciòn y Documentaciòn CientìficaSIGLEESSpai

Fotocatálisis: nanomateriales para combatir la contaminación y obtener energía

The photocatalytic process, based on the electron-hole pairs generation, separation and migration when a semiconductor is irradiated, has multiple sustainable applications for environmental (water, urban air, soil) depollution treatment, synthesis of added-value compounds or energy production, using solar radiationEl proceso fotocatalítico, basado en la generación, separación y migración de pares electrón-hueco cuando se irradia un semiconductor, tiene múltiples aplicaciones para tratamientos de descontaminación del medio ambiente (aguas, aire urbano, suelos), síntesis de compuestos de alto valor añadido u obtención de energía mediante celdas solares. Todo ello de forma sostenible al utilizar la radiación sola

Oxidación selectiva de metano a formaldehído sobre catalizadores vanadio-sílice

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Química Inorgánica. Fecha de lectura: 11-7-199

An approach on the comparative behavior of chloro / nitro substituted phenols photocatalytic degradation in water

This Accepted Manuscript will be available for reuse under a CC BY-NC-ND licence after 24 months of embargo periodThe study of position and number of substituents on the photocatalytic removal of some mono-, di- and tri-, chloro- and nitrophenols, as well as more known initial TOC concentration effect, has revealed the noteworthy impact on the process efficiency. Despite the complex effect of multiple substituents directing the HO attack to their preferential positions, Hammett constant could be used to predict photocatalytic degradation performance. TiO2 P25 was able to mineralize initial TOC concentrations up to 25-50 mg·L-1. Higher TOC concentrations constitute a drawback and drive to residual parent pollutants and organic by-products, which become more important when raising the starting TOC loading. Increasing the number of chloro- or nitro- groups in the aromatic ring does not imply higher ecotoxiticy values; contrarily, the position of the substituent can lead to significant differences. TOC conversion values are hardly affected by the substituent position, but by the number of groups in the organic molecule, probably due to steric hindrance. The formation of chloride and nitrogen inorganic ions, inherent to photocatalytic degradation, fulfills the Cl and N mass balances. Finally, the effect of number, position and electronic nature of substituents on contaminant initial photocatalytic degradation rates was studied by corresponding Hammett constant correlationsThis work has been supported by the Spanish Plan Nacional de I+D+i through the project CTM2015-64895-R. Alvaro Tolosana-Moranchel thanks to Ministerio de Educación, Cultura y Deporte for his FPU grant (FPU14/01605). The authors are also grateful to Evonik Company for TiO2 sampl

2-Azapinanes: Aza Analogues of the Enantiomeric Pinyl Carbocation Intermediates in Pinene Biosynthesis

The enantiomeric 2-azapinanes, aza analogues of the pinyl carbocation intermediates in pinene biosynthesis, were synthesized from (−)- and (+)-cis-pinonic acids. The individual reactions in the 5-step sequence were Beckmann rearrangement of the pinonic acid oximes, cyclization to the N-acetyl lactams, hydrolysis to the NH-lactams, N-methylations, and LiAlH4 reductions. The anti stereochemistry of the N-methyl groups in the salts with respect to the gem-dimethyl bridge was established by NOE measurements and by X-ray diffraction analysis

2-Azapinanes: Aza Analogues of the Enantiomeric Pinyl Carbocation Intermediates in Pinene Biosynthesis

The enantiomeric 2-azapinanes, aza analogues of the pinyl carbocation intermediates in pinene biosynthesis, were synthesized from (−)- and (+)-cis-pinonic acids. The individual reactions in the 5-step sequence were Beckmann rearrangement of the pinonic acid oximes, cyclization to the N-acetyl lactams, hydrolysis to the NH-lactams, N-methylations, and LiAlH4 reductions. The anti stereochemistry of the N-methyl groups in the salts with respect to the gem-dimethyl bridge was established by NOE measurements and by X-ray diffraction analysis