Substrate-dependent stereochemical course of the (Z)-13-desaturation catalyzed by the processionary moth multifunctional desaturase.

The stereochemical course of the Delta13 desaturation involved in the biosynthesis of Thaumetopoea pityocampa sex pheromone was studied using stereotopically labeled and tagged palmitic acids as metabolic probes. In the synthetic pathway, a functionalized acetylene common synthon was used for introducing the four deuterium tags. Further coupling of the tetradeuterated synthon to the appropriated alkynol and a double chemoenzymatic strategy to resolve the alcohol functionality allowed one to obtain the enantiomerically enriched probes used in the mechanistic studies.

A multifunctional desaturase involved in the biosynthesis of the processionary moth sex pheromone.

The sex pheromone of the female processionary moth, Thaumetopoea pityocampa, is a unique C16 enyne acetate that is biosynthesized from palmitic acid. Three consecutive desaturation reactions transform this saturated precursor into the triunsaturated fatty acyl intermediate: formation of (Z)-11-hexadecenoic acid, acetylenation to 11-hexadecynoic acid, and final Delta(13) desaturation to (Z)-13-hexadecen-11-ynoic acid. By using degenerate primers common to all reported insect desaturases, a single cDNA sequence was isolated from total RNA of T.

Synthesis and use of deuterated palmitic acids to decipher the cryptoregiochemistry of a Delta13 desaturation.

The synthesis of two hexadeuterated palmitic acids differing in the position of the diagnostic labels, and their use to decipher the cryptoregiochemistry of a Delta13 desaturation are described. A dithiane and a triple bond functionalities were used to introduce the diagnostic (C13 or C14) and tagging (C8 and C9) labels, respectively, in the palmitic acid skeleton. Using these probes, the cryptoregiochemistry of the Delta13 desaturation involved in the biosynthesis of Thaumetopoea pityocampa sex pheromone was studied by means of kinetic isotope effect determinations.

Synthesis and use of probes to investigate the cryptoregiochemistry of the first animal acetylenase.

Thaumetopoea pityocampa pheromone glands contain an unusual Delta(11) acetylenase that produces an alkynoic fatty acid intermediate in the sex pheromone biosynthetic pathway of this species. In this article, we describe the synthesis and use of the deuterated (Z)-11-hexadecenoic acid probes required to decipher the cryptoregiochemistry of this enzyme. The label in the olefinic bonds was introduced by Wittig reaction between the appropriate deuterated reagents.

Biosynthesis of 10,12-dienoic fatty acids by a bifunctional Delta11 desaturase in Spodoptera littoralis.

In the biosynthetic pathway of Spodoptera littoralis sex pheromone, (E,E)-10,12-tetradecadienoic acid is produced from (Z)-11-tetradecenoic acid by desaturation and concomitant migration of the precursor double bond. With the aim of identifying the enzyme involved in this biotransformation, yeast Deltaelo1/Deltaole mutants, which are both elongase 1 and Delta9 desaturase-deficient, were transformed with the S. littoralis Delta11 desaturase gene using a Cu+2 inducible expression vector.

Expression and evolution of delta9 and delta11 desaturase genes in the moth Spodoptera littoralis.

Desaturation of fatty acids is a key reaction in the biosynthesis of moth sex pheromones. The main component of Spodoptera littoralis sex pheromone blend is produced by the action of Delta11 and Delta9 desaturases. In this article, we report on the cloning of four desaturase-like genes in this species: one from the fat body (Sls-FL1) and three (Sls-FL2, Sls-FL3 and Sls-FL4) from the pheromone gland.

Synthesis and use of stereospecifically deuterated analogues of palmitic Acid to investigate the stereochemical course of the delta11 desaturase of the processionary moth.

Thaumetopoea pityocampa pheromone glands contain desaturases that, after several sequential reactions from palmitic acid, catalyze the formation of a unique enyne fatty acid, which is the immediate sex pheromone precursor. In this article, we describe the synthesis of different stereospecifically deuterium-labeled and isotopically tagged palmitic acid probes needed to decipher the stereochemical course of the T. pityocampa Delta(11) desaturase.

Synthesis of deuterated fatty acids to investigate the biosynthetic pathway of disparlure, the sex pheromone of the gypsy moth, Lymantria dispar.

The preparation and characterization of a series of deuterium-labeled intermediates used in the study of the biosynthetic pathway for disparlure, the sex pheromone of Lymantria dispar, is reported. The synthetic route starts with propargyl alcohol, and the deuterium atoms are introduced by deuteration of an alkyne precursor in the presence of Wilkinson's catalyst. The olefinic bond was created by the Wittig reaction of a suitable aldehyde with a common tetradeuterated phosphonium ylide intermediate.

Inhibition of the acyl-CoA desaturases involved in the biosynthesis of Spodoptera littoralis sex pheromone by analogs of 10,11-methylene-10-tetradecenoic acid.

The desaturase inhibitory activity of the cyclopropenyl alcohols 9,10-methylene-9-tetradecen-1-ol (9-MTOL), 10,11-methylene-10-tetradecen-1-ol (10-MTOL) and 11,12-methylene-11-tetradecen-1-ol (11-MTOL), which are structural analogs of 10,11-methylene-10-tetradecenoic acid (10-MTA), is reported. At equimolar ratios with respect to the different substrates, the three compounds completely inhibited the three desaturation reactions involved in the biosynthesis of Spodoptera littoralis sex pheromone. The dose-dependence of inhibition was determined for 10-MTA and its alcohol derivative.

Field trials with the synthetic sex pheromone of the oak processionary moth Thaumetopoea processionea.

The biological activity of synthetic (Z,Z)-11,13-hexadecadienyl acetate, the major pheromone component found in female gland extracts of the oak processionary moth Thaumetopoea processionea, was evaluated in field trials. Traps baited with 10 mg of the chemical efficiently attracted a large number of males provided they were placed in the upper crown region of the oaks. Devices positioned 10-15 m high in the trees attracted significantly more males than those traps installed at 2 or 6-8 m above the ground.

Synthesis of fluorinated analogs of myristic acid as potential inhibitors of Egyptian armyworm (Spodoptera littoralis) delta11 desaturase.

To study the activity of the different desaturases present in the pheromone biosynthetic pathway of the Egyptian armyworm, Spodoptera littoralis, we prepared a series of mono- and gem-difluorinated analogs of myristic acid with halogen substitution at the C8-C11 positions of the aliphatic chain via specifically positioned dithiane precursors. Thus, transformation of dithianes by treatment with N-bromosuccinimide in the presence of H2O followed by reduction with LiAlH4 afforded the appropriate alcohols, which reacted with diethylaminosulfur trifluoride to give rise to the corresponding monofluoroderivative intermediates. Alternatively, the introduction of the gem-difluoro functionality was carried out by reaction of the appropriate dithiane intermediate with 1,3-dibromo-5,5-dimethylhydantoin in the presence of HF/pyridine.

Novel chemoenzymatic strategy for the synthesis of enantiomerically pure secondary alcohols with sterically similar substituents.

A novel chemoenzymatic strategy for the synthesis of enantiomerically pure secondary alcohols with sterically similar substituents is described. The key step is the kinetic lipase-catalyzed resolution of racemic mixtures of substituted propargylic alcohols. The efficiency of this new approach was tested in the preparation of the corresponding enantiomers of 1,11-hexadecandiol derivatives ((R)-5 and (S)-5).

Sex pheromone of the oak processionary moth Thaumetopoea processionea. Identification and biological activity.

The sex pheromone of the oak processionary moth Thaumetopoea processionea has been characterized from female gland extracts as a mixture of (Z,Z)-11,13-hexadecadienyl acetate (1), (E,Z)-11,13-hexadecadienyl acetate (3) and (Z,Z)-11,13-hexadecadienol (2) in 88:7:5 ratio. The amount of the major compound 1 was 20-30 ng/gland. No trace of (Z,Z)-11,13-hexadecadienal was found in the extract, and therefore, T.

Enzymatic desaturation of fatty acids: delta11 desaturase activity on cyclopropane acid probes.

The formation of methylenecyclopropanes by enzymatic desaturation of 11-cyclopropylundecanoic acid (1) and its disubstituted derivatives cis- and trans-3-5 has been investigated using the Delta(11) desaturase of Spodoptera littoralis as model enzyme. Gas chromatography coupled to mass spectrometry analyses of methanolyzed lipidic extracts from tissues incubated with each probe revealed that all the cyclopropyl fatty acids were transformed into the corresponding 11-cyclopropylidene acids, except for compound trans-5 (5b), which was not desaturated at C11. The formation of methylenecyclopropane 9 as the only reaction product from 1 indicates that a potential radical intermediate is too short-lived to allow rearrangement reactions.

[Modification of the analytical method for rotenoids in plants].

Rotenoids are the active ingredients of some botanical insecticides and prospective candidates as anticancer agents. The proper isolation and determination of rotenoids in plants is of great importance for their further research and development. However, the HPLC method available for this purpose was developed particularly for the detection and determination of rotenone, so it appears to be unsuitable for the analysis of other rotenoids such as deguelin, elliptone and their analogues.

A new member of the PBAN family in Spodoptera littoralis: molecular cloning and immunovisualisation in scotophase hemolymph.

In this article, we report evidence suggesting that the immunoreactive factor previously detected in Spodoptera littoralis scotophase hemolymph is PBAN, which supports a humoral route of the hormone to the pheromone gland. Western blot after native-PAGE of prepurified scotophase hemolymph extracts yielded an immunoreactive band with the same mobility as S. littoralis Br-SOG factor and the expected mobility for a noctuid PBAN.

Attractant volatiles released by female and male Triatoma infestans (Hemiptera: Reduviidae), a vector of chagas disease: chemical analysis and behavioral bioassay.

Volatiles emitted by male and female T infestans before and during copula were collected on Porapak-Q filters, desorbed with dichloromethane, and analyzed by gas chromotography and gas chromatography-mass spectrometry after confirmation of attractiveness in an arena bioassay. Chemical analysis confirmed the presence of (R,S) -2- and 3-methylbutan-1-ol in a 2:1 ratio; short chain acids (ethanoic to nonanoic acid); long chains acids decanoic to (Z)-9-octadecenoic acid; aliphatic aldehydes (hexanal to nonanal), benzaldehyde and dipropylsulphide from insects in copula. Electroantennographic studies conducted with a homologous series of aliphatic aldehydes on female and male T infestans showed that, for a given dose, EAG responses elicited from both sexes increased with increased chain length up to nonanal, after which EAG-activity declined.

Stereospecificity of an enzymatic monoene 1,4-dehydrogenation reaction: conversion of (Z)-11-tetradecenoic acid into (E,E)-10,12-tetradecadienoic acid.

In this article, we report the first stereochemical study of an enzymatic 1,4-dehydrogenation reaction, namely, the transformation of (Z)-11-tetradecenoic acid into (E,E)-10,12-tetradecadienoic acid, involved in the sex pheromone biosynthesis of the moth Spodoptera littoralis. The investigation was carried out using the labeled substrates (R)-[10-(2)H]- and (S)-[10-(2)H]-tridecanoic acids ((R)-2 and (S)-2, respectively) and (R)-[2,2,3,3,13-(2)H(5)]- and (S)-[2,2,3,3,13-(2)H(5)]-tetradecanoic acids ((R)-1 and (S)-1, respectively). Probes (R)-2 and (S)-2 were prepared as described in a previous article.

Development and evaluation of an immunoassay for biological monitoring chlorophenols in urine as potential indicators of occupational exposure.

Trichlorophenols (TCP) eliminated by the urine can be considered as potential biomarkers of exposure of many chemicals (chlorophenols, chlorophenoxy acid herbicides, prochloraz, lindane, hexachlorobenzene, etc). High-throughput screening methods are necessary to carry out efficient monitoring programs that may help to prevent certain occupational health diseases. For this purpose, an indirect enzyme-linked immunosorbent assay (ELISA) for 2,4,6-trichlorophenol detection has been developed using polyclonal antisera raised against 3-(3-hydroxy-2,4,6-trichlorophenyl)propanoic acid (hapten 5) covalently coupled by the mixed anhydride (MA) method to keyhole limpet hemocyanin (KLH).

Trapping of Cyclopropenyl Radicals by 5,5-Dimethyl-1-pyrroline--oxide.

The possible generation of cyclopropenyl radicals by ultraviolet irradiation of different cyclopropenyl derivatives in fluid solution and in the presence of 5,5-dimethyl-1-pyrroline--oxide (DMPO) as spin trap has been detected by electron paramagnetic resonance. The spectra consist of doublets of triplets in which the β-hydrogen splitting is larger than that of the nitrogen, in good agreement with data reported in the literature for other DMPO adducts.This methodology is unprecedented in the study of these transient radical species, and these results suggest the participation of cyclopropenyl radicals in the photosensitized decarboxylation of -(2-cyclopropenylcarbonyloxy)phthalimides.