Minor lignans of Piper clusii

Further investigations on the petrol extract of Piper clusii have afforded four more new lignans.These are 2S,3R,4R,2-ethoxy-3-(3,4,5-trimethoxyphenyl)methyl 4-(1,3-benzodioxol-5-yl) methyl tetrahydrofuranol; 3R,4R,bis-3,4-(3,4,5-trimethoxyphenyl) methyl tetrahydrofuran-2-one; 2R,3R,2-(7-methoxy-1,3-benzodioxol-5-yl) methyl 3-(3,4,5-trimethoxyphenyl)methyl butan-1,4-diol and 2R,3R,2-(1,3-benzodioxol-5-yl)methyl 3-(3,4,5-trimethoxyphenyl) methyl butan-1,4-diol. This is the first report of these compounds from a natural source.     

Engineering of Ralstonia eutropha H16 for Autotrophic and Heterotrophic Production of Methyl Ketones

Ralstonia eutropha is a facultatively chemolithoautotrophic bacterium able to grow with organic substrates or H₂ and CO₂ under aerobic conditions. Under conditions of nutrient imbalance, R. eutropha produces copious amounts of poly[(R)-3-hydroxybutyrate] (PHB). Its ability to utilize CO₂ as a sole carbon source renders it an interesting new candidate host for the production of renewable liquid transportation fuels. We engineered R. eutropha for the production of fatty acid-derived, diesel-range methyl ketones. Modifications engineered in R. eutropha included overexpression of a cytoplasmic version of the TesA thioesterase, which led to a substantial (>150-fold) increase in fatty acid titer under certain conditions. In addition, deletion of two putative β-oxidation operons and heterologous expression of three genes (the acyl coenzyme A oxidase gene from Micrococcus luteus and fadB and fadM from Escherichia coli) led to the production of 50 to 65 mg/liter of diesel-range methyl ketones under heterotrophic growth conditions and 50 to 180 mg/liter under chemolithoautotrophic growth conditions (with CO₂ and H₂ as the sole carbon source and electron donor, respectively). Induction of the methyl ketone pathway diverted substantial carbon flux away from PHB biosynthesis and appeared to enhance carbon flux through the pathway for biosynthesis of fatty acids, which are the precursors of methyl ketones.     

Selective ϖ-1 oxidation of fatty acids by CYP147G1 from Mycobacterium marinum

Background

Cyp147G1 is one of 47 cytochrome P450 encoding genes in Mycobacterium marinum M, a pathogenic bacterium with a high degree of sequence similarity to Mycobacterium tuberculosis and Mycobacterium ulcerans. Cyp147G1 is one of only two of these cyp genes which are closely associated with a complete electron transfer system.

Interactions of lipoidal materials and a pyridazinone inhibitor of chloroplast development

Formation of chloroplast pigments was inhibited, and free fatty acids accumulated in mustard (Brassica juncea [L.] Coss.) cotyledons and in barley (Hordeum vulgare L.) first leaves developed after treatment with 4-chloro-5- (dimethylamino)-2- (α, α, α-trifluoro-m-tolyl) -3 (2H) -pyridazinone. The inhibitor reduced the amount of fatty acids found in polar lipids (galactolipids) of barley chloroplasts and increased the amount in nonpolar lipids while having little effect on total content of bound fatty acids. The inhibition of chlorophyll formation was circumvented by D-α-tocopherol acetate, phytol, farnesol, and squalene, and by unsaturated fatty acids and their methyl esters. The protective action can be explained partially by an interaction external to the plant whereby 4-chloro-5- (dimethylamino) -2- (α, α, α-trifluoro-m-tolyl) -3 (2H) -pyridazinone partitioned out of the aqueous phase and into the lipid phase, thus limiting availability of the inhibitor to plants. However, the amount of inhibitor reaching the cotyledons of tocopherol-protected mustard seedlngs was still in excess of the amount necessary to cause white foliage, but it failed to produce the effect. Tocopherol treatment did not prevent the 4-chloro-5- (dimethylamino) -2- (α, α, α-trifluoro-m-tolyl) -3 (2H) -pyridazinone-induced buildup of fatty acids in mustard cotyledons but did partially circumvent the effect in barley leaves. The amount of linolenic acid relative to linoleic acid was reduced in barley leaves and chloroplasts by 4-chloro-5- (dimethylamino) -2- (α, α, α-trifluoro-m-tolyl) -3 (2H) -pyridazinone action and this effect was circumvented by tocopherol.     

Reaktionen von kohlenhydraten mit dichlormethylendimethylammoniumchlorid: ein neuer weg zu 2-chlor-2-desoxy-, und 3-chlor-3-desoxyhexose-, und 3-chlor-3-desoxypentosederivaten

Treatment of methyl 4,6-O-benzylidene-α-D-mannopyranoside with dichloromethylenedimethylammonium chloride gave methyl 4,6-O-benzylidene-3-chloro-3-deoxy-2-(N,N-dimethylcarbamoyl)-α-D-altropyranoside and methyl 4,6-O-benzy]idene-2-chloro-2-deoxy-3-(N,N-dimethylcarbamoyl)-α-D-glucopyranoside. Methyl 4,6-O-benzylidene-α-D-allopyranoside gave under analogous conditions the corresponding 2-chloro-3-(N,N-dimethylcarbamoyl)-α-D-altrose and 3-chloro-2-(N,N-dimethylcarbamoyl)-α-D-glucose derivatives. Methyl 5-O-benzyl-α,β-D-ribofuranoside and methyl 5-O-methyl-β-D-ribofuranoside gave only the corresponding methyl 3-chloro-2-(N,N-dimethylcarbamoyl)-α-D-xylofuranoside derivatives.     

Stimulation of nitrogen removal in the rhizosphere of aquatic duckweed by root exudate components

Plants can stimulate bacterial nitrogen (N) removal by secretion of root exudates that may serve as carbon sources as well as non-nutrient signals for denitrification. However, there is a lack of knowledge about the specific non-nutrient compounds involved in this stimulation. Here, we use a continuous root exudate-trapping system in two common aquatic duckweed species, Spirodela polyrrhiza (HZ1) and Lemna minor (WX3), under natural and aseptic conditions. An activity-guided bioassay using denitrifying bacterium Pseudomonas fluorescens showed that crude root exudates of the two species strongly enhanced the nitrogen-removal efficiency (NRE) of P. fluorescens (P < 0.05) under both conditions. Water-insoluble fractions (F) obtained under natural conditions stimulated NRE to a significant extent, promoting rates by about 30 %. Among acidic, neutral and basic fractions, a pronounced stimulatory effect was also observed for the neutral fractions from HZ1 and WX3 under both conditions, whereas the acidic fractions from WX3 displayed an inhibitory effect. Analysis of the active fractions using gas chromatography/mass spectrometry (GC/MS) revealed that duckweed released fatty acid methyl esters and fatty acid amides, specifically: methyl hexadecanoate, methyl (Z)-7-hexadecenoate, methyl dodecanoate, methyl-12-hydroxystearate, oleamide, and erucamide. Methyl (Z)-7-hexadecenoate and erucamide emerged as the effective N-removal stimulants (maximum stimulation of 25.9 and 33.4 %, respectively), while none of the other tested compounds showed stimulatory effects. These findings provide the first evidence for a function of fatty acid methyl esters and fatty acid amides in stimulating N removal of denitrifying bacteria, affording insight into the “crosstalk” between aquatic plants and bacteria in the rhizosphere.     

Peracid oxidation of methyl esters of γ-hydroxy-α,β-unsaturated fatty acids

Oxidation of methyl 4-hydroxy-trans-2-octadecenoate (I) with m-chloroperbenzoic acid gave a rearrangement product characterized as methyl 3-keto-4-hydroxyoctadecanoate (II). Chromium trioxide-pyridine oxidation and sodium borohydride reduction of (II) yielded methyl 3,4-diketooctadecanoate (III) and 1,3,4-octadecane-triol (IV), respectively. The structures of these fatty acid derivatives are determined by chemical methods, elemental analysis, infrared (IR) and proton nuclear magnetic resonance spectroscopy (NMR) as well as by gas chromatography/mass spectrometry (GC-MS). The intramolecular isomerization of epoxy to keto group provides a useful method for the preparation of long-chain β,γ-ketol acids.     

Cyclic glycolipids from glandular trichome exudates of Cerastium glomeratum

Fourteen cyclic glycolipids, named glomerasides A–N, have been isolated from the glandular trichome exudate of Cerastium glomeratum (Caryophyllaceae). Their structures were determined by spectroscopic analysis of the glycolipids, as well as by application of the Ohrui–Akasaka method to the fatty acid methyl esters derived from the glycolipids and GCMS studies of trimethylsilyl ether derivatives of the methyl esters. The various glomerasides have a glycosidic linkage between the anomeric hydroxy group of the glucose and the C-11, C-10 or C-9 positions of the docosanoyl moiety. They also contained an ester linkage between the C-6 hydroxy group of the glucose ring and the carboxyl group of the oxygenated fatty acid to form their macrocyclic structures. The glucose moiety was optionally acetylated and/or malonylated at the C-2 or C-3 hydroxy groups. Among these compounds, the 1,6′-cyclic ester of 11(R)-(2-O-acetyl-β-d-glucopyranosyloxy)docosanoic acid (glomeraside D) was the most abundant (25%).     

Biodiesel production from rice bran oil and supercritical methanol

In this study, production of biodiesel from low cost raw materials, such as rice bran and dewaxed-degummed rice bran oil (DDRBO), under supercritical condition was carried out. Carbon dioxide (CO₂) was employed as co-solvent to decrease the supercritical temperature and pressure of methanol. The effects of different raw materials on the yield of biodiesel production were investigated. In situ transesterification of rice bran with supercritical methanol at 30 MPa and 300 °C for 5 min was not a promising way to produce biodiesel because the purity and yield of fatty acid methyl esters (FAMEs) obtained were 52.52% and 51.28%, respectively. When DDRBO was reacted, the purity and yield were 89.25% and 94.84%, respectively. Trans-FAMEs, which constituted about 16% of biodiesel, were found. They were identified as methyl elaidate [trans-9], methyl linoleaidate [trans-9, trans-12], methyl linoleaidate [cis-9, trans-12], and methyl linoleaidate [trans-9, cis-12]. Hydrocarbons, which constituted about 3% of the reaction product, were also detected.     

Synthesis of carbon-11-labeled 5-HT6R antagonists as new candidate PET radioligands for imaging of Alzheimer’s disease

Carbon-11-labeled serotonin (5-hydroxytryptamine) 6 receptor (5-HT₆R) antagonists, 1-[(2-bromophenyl)sulfonyl]-5-[¹¹C]methoxy-3-[(4-methyl-1-piperazinyl)methyl]-1H-indole (O-[¹¹C]2a) and 1-[(2-bromophenyl)sulfonyl]-5-methoxy-3-[(4-[¹¹C]methyl-1-piperazinyl)methyl]-1H-indole (N-[¹¹C]2a), 5-[¹¹C]methoxy-3-((4-methylpiperazin-1-yl)methyl)-1-(phenylsulfonyl)-1H-indole (O-[¹¹C]2b) and 5-methoxy-3-((4-[¹¹C]methylpiperazin-1-yl)methyl)-1-(phenylsulfonyl)-1H-indole (N-[¹¹C]2b), 1-((4-isopropylphenyl)sulfonyl)-5-[¹¹C]methoxy-3-((4-methylpiperazin-1-yl)methyl)-1H-indole (O-[¹¹C]2c) and 1-((4-isopropylphenyl)sulfonyl)-5-methoxy-3-((4-[¹¹C]methylpiperazin-1-yl)methyl)-1H-indole (N-[¹¹C]2c), 1-((4-fluorophenyl)sulfonyl)-5-[¹¹C]methoxy-3-((4-methylpiperazin-1-yl)methyl)-1H-indole (O-[¹¹C]2d) and 1-((4-fluorophenyl)sulfonyl)-5-methoxy-3-((4-[¹¹C]methylpiperazin-1-yl)methyl)-1H-indole (N-[¹¹C]2d), were prepared from their O- or N-desmethylated precursors with [¹¹C]CH₃OTf through O- or N-[¹¹C]methylation and isolated by HPLC combined with SPE in 40–50% radiochemical yield, based on [¹¹C]CO₂ and decay corrected to end of bombardment (EOB). The radiochemical purity was >99%, and the molar activity (MA) at EOB was 370–740?GBq/μmol with a total synthesis time of ～40-min from EOB.     

Esters of trehalose from Corynebacterium diphtheriae: A modified purification procedure and studies on the structure of their constituent hydroxylated fatty acids

The purification procedure of 6,6′-diesters of trehalose from Corynebacterium diphtheriae was modified and the isolated substance was analysed by mass spectrometry as its permethylated derivative. The fatty acid moiety released from the glycolipid after alkaline hydrolysis was studied by mass spectral analysis of the O-methylated and O-acetylated methyl ester derivatives. By argentation thin-layer chromatography, three species of O-acetylated methyl esters were recognized, corresponding to saturated, mono-unsaturated and di-unsaturated α-branched-β-hydroxylated fatty acids. The double bond was located by ozonolysis of the O-acetylated methyl ester derivatives, by gas chromatography of the reaction product and mass spectrometry of the effluent from the gas chromatograph. The main components of each species of α-branched-β-hydroxylated fatty acids found in the gly colipid fraction of C. diphtheriae were 2-tetradecyl-3-hydroxyoctadecanoic acid (C₃₂H₆₄O₃, corynomycolic acid), 2-tetradecyl-3-hydroxy-11-octadecenoic acid (C₃₂H₆₂O₃, corynomycolenic acid), 2-tetradec-7′-enyl-3-hydroxy octadecanoic acid (C₃₂H₆₂O₃) and 2-tetradec-7′-enyl-3-hydroxy-11-octadecenoic acid (C₃₂H₆₀O₃, corynomycoldienic acid). The glycolipid fraction from C. diphtheriae is obviously a complex mixture of 6,6′-diesters of trehalose.     

Fatty Acid Profiling of Commercially Important Raw and Boiled Seer Fish <Emphasis Type="Italic">Scomberomorus commerson</Emphasis> (Lacepede, 1800) (Scombridae,Teleostei, Pisces)

The present study aims to determine the fatty acid profiling of commercially important fresh and boiled Scomberomorus commerson. Fatty acids in fresh and boiled fish were separated and quantitatively determined by gas chromatography–mass spectrophotometer using standard methods. The findings revealed that the predominant fatty acids in fresh S. commerson were octadecanoic acid methyl ester, octanoic acid, 1,2-benzenedicarboxylic acid and 3-cyclopentylpropionic acid representing respectively 45.91, 5.69, 6.75 and 8.65% of total fatty acids. Boiled S. commerson showed predominant changes in their fatty acid profiles. In the omega-3 and omega-6 families the dominant fatty acids were doconexent, 3-cyclopentylpropionic acid, octadeconoic acid methyl ester and Hexadecane representing respectively 3.87, 12.08, 44.26 and 3.11% of total fatty acids. After boiling, some fatty acids present in fresh fish are damaged and formed new fatty acids which belonged to ω-3 polyunsaturated fatty acids (PUFA). Boiling increased the concentration of PUFAs from 73.25 to 80.37% of total fatty acids and also formed new fatty acids.     

Dihydrozeatin: An improved synthesis and resolution of both isomers

(±)-Dihydrozeatin was synthesized in a 3-step synthesis by: (1) a Michael condensation of methyl methacrylate with nitromethane to give (±) - methyl 2 - methyl - 4 - nitrobutyrate, which was (2) reduced to (±) - 4 -amino - 2 - methylbutan - 1 - ol and (3) reaction of the aminoalcohol with 6-chloropurine. Hydrolysis of racemic nitroester gave (±) - 2 - methyl - 4 - nitrobutyric acid, which was resolved by means of (+) - and (-) -α -methylbenzylamine salts. Conversion of the salts to the corresponding methyl esters and subsequent reductions yielded optically active 4 - amino 2 - methylbutan - 1 - ols. Examination of the NMR spectra of the resolved methyl 2 - methyl - 4 - nitrobutyrates in the presence of a chiral shift reagent established their optical purities to be greater than 98%. The specific rotations at 589 nm of theS- (?) andR- (+)- dihydrozeatins derived from optically active butanols were appreciably lower than previously reported. Application of the Drude equation to ORD values from 320 to 589 nm verified the low 589 nm rotations of the dihydrozeatin enantiomers. The biological activities of (R), (S) and (R,S) dihydrozeatins in the betacyanin stimulation assay withAmaranthus parallel the activities found in other cytokinin bioassays.     

Uropygial gland lipids of penguins

The uropygial gland secretion of penguins (Sphenisciformes) is a complex mixture of monoester waxes. The wax fatty acids of Pygoscelis and Eudyptes are predominantly 3-methyl- and 3, x-dimethyl-substituted, whereas 2- and 4-monomethyl-, or 2, x-as well as 4, x-dimethyl-substituted acids predominate in the case of Spheniscus. The wax alcohols are very similar within all three species and are mainly composed of mono- and dimethyl-substituted compounds with branches predominantly in the middle or near the methyl end of the carbon chain. From these data, it is concluded that, chemotaxonomically, the penguins are closely related to the tubenoses (Procellariiformes).     

Structural characterization of saturated branched chain fatty acid methyl esters by collisional dissociation of molecular ions generated by electron ionization

Saturated branched chain fatty acids (BCFA) are present as complex mixtures in numerous biological samples. The traditional method for structure elucidation, electron ionization (EI) mass spectrometry, sometimes does not unambiguously enable assignment of branching in isomeric BCFA. Zirrolli and Murphy (Zirrolli , J. A. , and R. A. Murphy. 1993. Low-energy tandem mass spectrometry of the molecular ion derived from fatty acid methyl esters: a novel method for analysis of branched-chain fatty acids. J. Am. Soc. Mass Spectrom. 4: 223–229.) showed that the molecular ions of four BCFA methyl ester (BCFAME) yield highly characteristic fragments upon collisional dissociation using a triple quadrupole instrument. Here, we confirm and extend these results by analysis using a tabletop 3-D ion trap for activated molecular ion EI-MS/MS to 30 BCFAME. iso-BCFAME produces a prominent ion (30-100% of base peak) for [M-43] (M-C₃H₇), corresponding to the terminal isopropyl moiety in the original iso-BCFAME. Anteiso-FAME yield prominent ions (20-100% of base peak) corresponding to losses on both side of the methyl branch, [M-29] and [M-57], and tend to produce more prominent m/z 115 peaks corresponding to a cyclization product around the ester. Dimethyl and tetramethyl FAME, with branches separated by at least one methylene group, yield fragment on both sides of the sites of methyl branches that are more than 6 C away from the carboxyl carbon. EI-MS/MS yields uniquely specific ions that enable highly confident structural identification and quantification of BCFAME.     

Synthèse de trithioorthocarbonates de pyranosides par thermolyse de bis(dithiocarbonates)

The thermal decomposition of methyl 4,6-O-benzylidene-2,3-di-O-[(methylthio)-thiocarbonyl]-α-d-glucopyranoside afforded methyl 4,6-O-benzylidene-2-thio-α-d-mannopyranoside 3-O,2-S-(S,S-dimethyl trithioorthocarbonate) and methyl 4,6-O-benzylidene-3-thio-α-d-allopyranoside 2-O,3-S-(S,S-dimethyl trithioorthocarbonate) in good yield. This decomposition can be generalized to 1,3-diols derived from sugars. Thus methyl 2,3-di-O-methyl-4,6-di-O-[(methylthio)thiocarbonyl]-α-d-glucopyranoside afforded in the same way the corresponding trithioorthocarbonates, following a regioselective process. The structures of these trithioorthocarbonates are confirmed by spectral and chemical proofs.     

Changes in Cellular Fatty Acid Composition of Cephalosporium acremonium during Cephalosporin C Production

Cephalosporium acremonium was cultivated in fermentation medium containing sucrose or methyl oleate as a carbon source for cephalosporin C production. The level of antibiotic production was 48 g of cephalosporin C per liter under optimum conditions when methyl oleate was used. The C_18:1 (oleic acid) methyl ester appeared to be utilized faster than the C_18:2 (linoleic acid) methyl ester in fermentation broth. Physiological characteristics of C. acremonium were investigated by determining the fatty acid composition of the total cellular free lipid. Significant changes in cellular fatty acid composition occurred during inoculum cultivation and fermentation. The percentage of C_18:1 increased from 19.1 to 38.5%, but the percentage of C_18:2 decreased from 56.7 to 36.1%, and there was an increase in pH during inoculum cultivation. The cellular fatty acid composition of C. acremonium grown in fermentation medium containing methyl oleate (methyl oleate medium) was significantly different from that in fermentation medium containing sucrose (sucrose medium). The major fatty acids detected were C_16:0 (palmitic acid), C_18:1, and C_18:2. In methyl oleate medium, the ratio of C_18:1 to C_18:2 increased from 0.34 to 1.37, while the cell morphology changed from hyphae to arthrospores and conidia. In contrast, in sucrose medium, the ratio of C_18:1 to C_18:2 decreased from 0.70 to 0.43, and most of the cells remained hyphal at the end of fermentation. We observed that hyphae contained a higher proportion of C_18:2 but arthrospores and conidia contained a higher proportion of C_18:1.     

Synthesis of 3-deoxy-d-arabino-2-heptulosonic acid 7-phosphate by phosphorylation of methyl (methyl 3-deoxy-d-arabino-heptulopyranosid)onate

3-Deoxy-d-arabino-2-heptulosonic acid 7-phosphate (5), the first committed intermediate in aromatic amino acid biosynthesis, has been synthesized in good yield by treatment of methyl (methyl 3-deoxy-d-arabino-2-heptulopyranosid)onate with diphenylphosphoric chloride under mild conditions to give the 7-diphenyl phosphate. Catalytic removal of the phenyl residues, followed by base-catalyzed hydrolysis resulted in formation of (methyl 3-deoxy-d-arabino-2-heptulopyranosid)onic acid dihydrogen 7-phosphate (4), which yielded a crystalline tris-(cyclohexylammonium) salt. Acid-catalyzed hydrolysis of 4 afforded 5, which was used to purify 3-dehydroquinate synthase.     

Novel nucleosides as potent influenza viral inhibitors

Influenza virus infection constitutes a significant health problem in need of more effective therapies. We have recently identified ((2R,3S,4R,5R)-3-acetoxy-5-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-fluoro-3,4-dimethyl-tetrahydrofuran-2-yl) methyl benzoate (18c) as a potent influenza virus inhibitor. We now here report the synthesis and evaluation of a series of C-3′ modified ribose nucleosides. These novel compounds were prepared, primarily by taking known ((2R,3R,4R)-3-benzoyloxy-4-fluoro-4-methyl-5-oxo-tetrahydrofuran-2-yl)methyl benzoate (1) and converting it in to C-3 keto sugar (7), reacting C-3 keto group with methyl magnesium bromide, followed by coupling these sugars with purine and pyrimidine bases. Anti influenza viral activity was determined by screening against both A and B viral strains.