Infrared and raman studies of the methyl cis undecenoates and of the methyl undecynoates

The infrared spectra (of CCl₄ solutions) and the Raman spectra (of the neat liquids) of the eight isomeric methyl cis-undecenoates, of methyl undec-10-enoate, and of the nine isomeric methyl undecynoates have been obtained. Conjugation of the double bond with the carbonyl group lowers the wavenumber value of ν(CC) by 8 cm^?1 from the mean value of 1657 cm^?1. Conjugation of the triple bond with the carbonyl group gives rise to a single Raman band due to ν(CC) at 2241 cm^?1 whereas the non-conjugated, non-terminal triple bond gives a Fermi resonance doublet at 2234 and 2293 cm^?1.     

Theoretical studies on the conformations of methyl glycopyranosides

The σ-charges on various atoms of methyl glycosides have been computed by using the MO-LCAO method of Del Re. The potential and free energies of methyl aldohexopyranosides and methyl aldopentopyranosides in their C1(d) and 1C(d) conformations have been calculated. Minimization of the energies of these conformations has been studied by suitably tilting the axial C-C and C-O bonds. Considerable release of strain is achieved when tilts of 4.5 and 3° are given to the axial hydroxymethyl and hydroxyl groups, respectively, that are involved in Hassel-Ottar effect. A tilt of 3° is also found necessary for the axial OMe group involved in the Hassel-Ottar effect. The calculated free-energy values are in accord with experimental ones, after adding a value of 0.8 kcal.mole^?1 for the anomeric effect of -OMe group. These studies predict that all of the methyl aldohexopyranosides, except methyl α-d- and methyl β-d-idopyranosides, favour the C1 conformation. On the other hand, the energy calculations also predict that, of the eight methyl aldopentopyranosides studied, only methyl α-d- and methyl β-d-xylopyranosides and methyl α-d -ribopyranoside favour the C1(d) conformation; for the other pentopyranosides, considerable amounts of both C1(d) and 1C(d) conformations are present in the equilibrium mixture. The calculated values of the percentage of α-anomer present in the equilibrium mixture agree fairly well with those obtained experimentally.     

Synthesis and properties of methyl 9,12,15-octadecatrienoate geometric isomers

The eight geometrically isomeric methyl 9,12,15-octadecatrienoates were prepared by using the Wittig reaction to couple cis- or trans-3-hexyenyltriphenylphosphonium bromide and methyl 12-oxo-cis- or trans-9-dodecenoate. Pairs of geometric triene isomers formed were separated by partial silver resin chromatography. Physical constants including melting points, percent trans by infrared, equivalent chain lengths (ECL), and ¹³C nuclear magnetic resonance (NMR) chemcial shifts are tabulated for the individual isomers.     

Proton and carbon-13 nuclear magnetic resonance studies on methyl (methyl d-galactosid)uronates and their per-O-acetyl derivatives

The ¹H- and ¹³C-n.m.r. spectra of the anomeric methyl (methyl d-galactosid)uronates, as well as the ¹H-n.m.r. spectra of their acetyl derivatives, were analyzed. The spectra of the unacetylated d-galactopyranosiduronates showed good correlation with those of the corresponding anomeric d-galactopyranuronic acids and their methyl esters, and with those of the anomeric methyl d-galactopyranosides. From the values of the chemical shifts and coupling constants, it was concluded that the anomeric methyl (methyl d-galactopyranosid)uronates and their corresponding peracetates are in the ⁴C₁(d) conformation. The chemical shifts in the ¹³C-n.m.r. spectra show good correlation with those of the methyl d-galactosides. The signals of the furanose derivatives appear at fields lower than those of the corresponding pyranose compounds.     

Enzymatic removal of a cephalosporin methyl ester blocking group

Over 7000 microorganisms were screened to find an enzyme source for the hydrolysis of a C₄ methyl ester blocking group on 7-aminodesacetoxycephalosporanic acid (7-ADCA). Only one culture, Streptomyces capillispira Mertz and Higgens nov. sp., produced an enzyme that catalysed the reaction. Enzyme synthesis in a defined mineral salts medium was repressed by NH₃ and amino acids. Under optimum fermentation conditions, the maximum rate of substrate hydrolysis was 6 × 10^?10 mol min^?1 mg^?1 cell. The enzyme was recovered from the mycelia and partially purified by gel filtration. Kinetic studies by pH-stat titration indicated that the pH optimum was 7.5–8.5, the temperature optimum was 25–30°C, and the substrate K_m value was 2.3 mg ml^?1. The reaction products, 7-ADCA and methanol, were weak competitive inhibitors of the enzyme with K₁ values of 6.63 and 0.188 mg ml^?1, respectively. The enzyme also hydrolysed cefaclor and cephalexin methyl esters but did not hydrolyse cephalosporin ethyl esters. With further improvements in enzyme yields and stability, enzymatic deblocking of cephalosporins could provide an alternative to chemical deblocking processes.     

Synthesis of deuterated methyl 6,9,12-octadecatrienoate geometric isomers

Methyl cis-6,cis-9,cis-12-octadecatrienoate-15,15,16,16-d₄ and the corresponding cis,cis,trans isomer were obtained by coupling hexyl-d₄-triphenylphosphonium bromide and methyl 12-oxo-cis-6,cis-9-dodecadienoate by the Wittig reaction. The deuterated phosphonium salt was prepared from 3-hexynol by catalytic deuteration of the corresponding tetrahydropyranyl ether and intermediate formation of the bromide. The dienoic aldehyde ester was obtained through the intermediate dioxanyl and dimethoxy derivatives from the Wittig coupling of methyl 9-oxo-cis-6-nonenoate with [2-(1,3-dioxan-2-yl)ethyl]-triphenylphosphonium bromide. The monoenoic aldehyde ester was prepared in a similar manner by the Wittig reaction between methyl 6-oxohexanoate and the dioxanylphosphonium salt. The saturated aldehyde ester was obtained, through several steps, from the ozonolysis of cyclohexene. Geometric isomers formed during each of the Wittig reactions were separated by silver resin chromatography. ¹³C Nuclear magnetic resonance chemical shifts for the compounds prepared are presented.     

Conformations of methyl 3,4-dideoxy-DL-glyc-3-enopyranosides

The conformational equilibrium (°H₁ ?¹H°) has been established for 26 diastereoisomeric methyl 3,4-dideoxy-DL-glyc-3-enopyranosides on the basis of the coupling constant J_1,2. The position of equilibrium depends on the interplay of steric and polar factors. The concept of the allylic effect is useful in explaining some conformational phenomena.     

Biosynthesis of methyl branched hydrocarbons of the German cockroach Blattella germanica (L.) (orthoptera,blattellidae)

The precursors and directionality of synthesis of the methyl branched cuticular hydrocarbons and the female contact sex pheromone, 3,11-dimethyl-2-nonacosanone, of the German cockroach, Blattella germanica, were investigated by radiotracer and carbon-13 NMR techniques. The amino acids [G-³H]valine, [4,5-³H]isoleucine and [3,4-¹⁴C₂]methionine labeled the hydrocarbon fraction in a manner indicating that the carbon skeletons of all three amino acids serve as the methyl branch group donor. The incorporation of [1,4-¹⁴C₂]- and [2,3-¹⁴C₂]succinates into the hydrocarbon and acylglycerol/polar lipid fractions indicated that succinate also served as a precursor to methylmalonyl-CoA. Carbon-13 NMR analyses showed that [1-¹³C]propionate labeled the carbon adjacent to the tertiary carbon, and, for the 3,x-dimethylalkanes, that carbon-4 and not carbon-2 was enriched. [1-¹³C]Acetate labeled carbon-2 of these hydrocarbons. This indicates that the methyl branching groups of the 3,x-dimethylalkanes were inserted early in the chain elongation process. [3,4,5-¹³C₃]Valine labeled the methyl, tertiary and carbon adjacent to the tertiary carbon of the methyl branched alkanes. Thus, the methyl branched hydrocarbon was formed by the insertion of methylmalonyl units derived from propionate, isoleucine, valine, methionine and succinate early in chain elongation.     

High titer methyl ketone production with tailored Pseudomonas taiwanensis VLB120

Methyl ketones present a group of highly reduced platform chemicals industrially produced from petroleum-derived hydrocarbons. They find applications in the fragrance, flavor, pharmacological, and agrochemical industries, and are further discussed as biodiesel blends. In recent years, intense research has been carried out to achieve sustainable production of these molecules by re-arranging the fatty acid metabolism of various microbes. One challenge in the development of a highly productive microbe is the high demand for reducing power. Here, we engineered Pseudomonas taiwanensis VLB120 for methyl ketone production as this microbe has been shown to sustain exceptionally high NAD(P)H regeneration rates. The implementation of published strategies resulted in 2.1 g L_aq⁻¹ methyl ketones in fed-batch fermentation. We further increased the production by eliminating competing reactions suggested by metabolic analyses. These efforts resulted in the production of 9.8 g L_aq⁻¹ methyl ketones (corresponding to 69.3 g L_org⁻¹ in the in situ extraction phase) at 53% of the maximum theoretical yield. This represents a 4-fold improvement in product titer compared to the initial production strain and the highest titer of recombinantly produced methyl ketones reported to date. Accordingly, this study underlines the high potential of P. taiwanensis VLB120 to produce methyl ketones and emphasizes model-driven metabolic engineering to rationalize and accelerate strain optimization efforts.     

The synthesis of 3-acetamido-2,3,5,6-tetradeoxy-5-fluoro-d,l-ribo-hexofuranose by the direct fluorination of methyl 3-acetamido-2,3,6-trideoxy-d,l-arabino-hexopyranoside (methyl N-acetyl-d,l-acosaminide)

3-Acetamido-2,3,5,6-tetradeoxy-5-fluoro-d,l-ribo-hexofuranose was synthesized by direct, fluorinative dehydroxylation of methyl 3-acetamido-2,3,6-trideoxy-d,l-arabino-hexopyranoside with sulfur tetrafluoride-hydrogen fluoride. The furanose form and the ribo configuration, indicated by ¹³C- and ¹H-n.m.r. spectroscopy, respectively, were confirmed by a single-crystal, X-ray diffraction study.     

Simultaneous degradation of the pesticides methyl parathion and chlorpyrifos by an isolated bacterial consortium from a contaminated site

The simultaneous degradation of the pesticide methyl parathion and chlorpyrifos was tested using a bacterial consortium obtained by selective enrichment from highly contaminated soils in Moravia (Medellin, Colombia). Microorganisms identified in the consortium were Acinetobacter sp, Pseudomonas putida, Bacillus sp, Pseudomonas aeruginosa, Citrobacter freundii, Stenotrophomonas sp, Flavobacterium sp, Proteus vulgaris, Pseudomonas sp, Acinetobacter sp, Klebsiella sp and Proteus sp. In culture medium enriched with each of the pesticides, the consortium was able to degrade 150 mg l⁻¹ of methyl parathion and chlorpyrifos in 120 h. When a mixture of 150 mg l⁻¹ of both pesticides was used the percentage decreased to 72% for methyl parathion and 39% for chlorpyrifos. With the addition of glucose to the culture medium, the consortium simultaneously degraded 150 mg l⁻¹ of the pesticides in the mixture. 4 treatments were carried out in soil that included the addition of glucose with microorganisms, the addition of sugar cane with microorganisms, microorganisms without nutrient addition and without the addition of any item. In the treatment in which glucose was used, degradation percentages of methyl parathion and chlorpyrifos of 98% and 97% respectively were obtained in 120 h. This treatment also achieved the highest percentage of reduction in toxicity, monitored with Vibrio fischeri.     

On the mechanism of action of methyl chymotrypsin

The properties of a-chymotrypsin methylated at histidine-57 were examined to explain the mechanism of this enzyme which is about 10⁵ times less active than chymotrypsin. Studies on the protein showed (i) an alteration in the acyl and leaving group specificity, (ii) decreased binding of some protein protease inhibitors by methyl chymotrypsin, (iii) lack of dimerization of methyl chymotrypsin at low pH, (iv) decreased stability of methyl chymotrypsin in urea, (v) a larger solvent deuterium isotope effect with methyl chymotrypsin, and (vi) decreased binding of a tetrahedral intermediate analog to methyl chymotrypsin. These properties suggest that while only subtle alterations occur in the active site upon methylation of His-57, the transition state and the tetrahedral intermediate are destabilized but not to the same extent. General base catalysis remains an integral feature of the hydrolytic mechanism of the modified chymotrypsin, and the base appears to be the methylated nitrogen of the imidazole moiety of His-57.     

Biosynthesis of propyl cannabinoid acid and its biosynthetic relationship with pentyl and methyl cannabinoid acids

Biosynthesis of propyl cannabinoid acid has been determined by in vitro incubation with a crude enzyme solution from three strains of Cannabis sativa using ¹⁴C-labelled cannabinoid acid. Biosynthetic relationships between methyl, propyl and pentyl cannabinoid acids have been demonstrated.     

The palladium(II) promoted hydrolysis of the methyl esters of glycyl-L-leucine,glycyl-L-alanine and L-alanylglycine

The palladium(II)-promoted hydrolysis of the methyl esters of glycyl-L-leucine, glycyl-L-alanine and L-alanylglycine have been studied at 25 °C and I=0.1 M in the pH range 4–5. At a 1:1 metal to ligand ratio the peptide esters act as tridentate ligands, donation occurring via the terminal amino group, the deprotonated amide nitrogen, and the carbonyl group of the ester. Due to the high Lewis acidity of Pd(II) rapid hydrolysis of the ester function by water and hydroxide ion occurs. Rate constants k_OH and k_H2O have been obtained for base hydrolysis and water hydrolysis of the coordinated peptide esters at 25 °C. The rate constants for base hydrolysis are 3.4 X 10⁶ M⁻¹ s⁻¹ (L-alaglyOMe), 6.4 X 10⁶ M⁻¹ s⁻¹ (gly-L-alaOMe) and 2.3 X 10⁷ M⁻¹ s⁻¹ (gly-L-leuOMe). Base hydrolysis of the coordinated peptide esters is at least 10⁶ times that of the free unprotonated ligand. Activation parameters have been obtained for both water and base hydrolysis of the Pd(II) complex of methyl L-alanylglycinate and possible mechanisms for the hydrolyses are considered.     

Synthesis and NMR characterization of the methyl esters of eicosapentaenoic acid monoepoxides

The activities of cytochrome P450-derived epoxide metabolites of omega-6 polyunsaturated fatty acids (PUFAs) in cellular homeostasis have generated considerable topical interest, but there is less information on the effects of omega-3 PUFA epoxides. Mass spectroscopic data on the epoxides of the omega-3 PUFA eicosapentaenoic acid (EPA) have been reported but the absence of corresponding NMR data currently hinders their biological assessment. In the present study five monoepoxy derivatives of EPA methyl ester were synthesized by treating EPA methyl ester with m-chloroperbenzoic acid. The individual regioisomers were purified by normal-phase chromatography and characterized by LC-MS/MS and a combination of NMR approaches including ¹H-, ¹³C-, ¹H-¹H-COSY, ¹H-¹³C-HSQC, and ¹H-¹³C-HMBC. The chromatographic properties for these monoepoxides were studied in normal-phase and reversephase-HPLC systems and the MS/MS fragmentation patterns using electrospray ionization were established. This paper also focuses on the NMR characterization of epoxide, olefinic and methylenic moieties and the complete assignments of the isomers.     

Unsaturated sugars. I. Decarboxylative elimination of methyl 2,3-di-O-benzyl-alpha-D-glucopyranosiduronic acid to methyl 2,3-di-O-benzyl-4-deoxy-beta-L-threo-pent-4-enopyranoside

Decarboxylative elimination of methyl 2,3-di-O-benzyl-α-D-glucopyranosiduronic acid (1) with N,N-dimethylformamide dineopentyl acetal in N,N-dimethylformamide gave methyl 2,3-di-O-benzyl-4-deoxy-β-L-threo-pent-4-enopyranoside (3). Debenzylation of 3 was effected with sodium in liquid ammonia to give methyl 4-deoxy-β-L-threo-pent-4-enopyranoside (4). Hydrogenation of 3 catalyzed by palladium-on-barium sulfate afforded methyl 2,3-di-O-benzyl-4-deoxy-β-L-threo-pentopyranoside (5), whereas hydrogenation of 3 over palladium-on-carbon gave methyl 4-deoxy-β-L-threo-pentopyranoside (6). An improved preparation of methyl 4,6-O-benzylidene-α-D-glucopyranoside is also described.     

Production of methyl sulfide and dimethyl disulfide from soil-incorporated plant materials and implications for controlling soilborne pathogens

Soil-incorporated plant materials have been associated with reduction in soilborne pathogens and diseases. Mechanisms of the biocidal actions are complex and not well understood. A glasshouse experiment, a non replicated field demonstration, and a field experiment were conducted to determine volatile compounds after incorporation of various plant species and their effect on pest control. Cabbage (Brassica oleracea), canola (Brassica rapa), kale (Brassica oleracea var. acephala), lettuce (Lactuca sativa var. valmaine), two mustard varieties -Caliente (Brassica juncea) and Green wave (Brassica juncea), two radish varieties - Oil seed (Raphanus sativus var. oleiformis) and Cherriette (Raphanus sativus), common rye (Secale cereale), and sorghum Sudan grass (Sorghum bicolor var. sudanese) were used in the glasshouse experiment. Caliente 199 mustard (Brassica hirta) was planted in the field demonstration and white mustard (Sinapis alba) was used in the field experiment. Fresh plant materials were chopped manually in the glasshouse experiment and mechanically in the field studies at the flowering stage before incorporation in natural field soils. In the glasshouse experiment, the equivalent biomass dry weight ranged from a minimum of 573 g?m^?2 for L. sativa var. valmaine to a maximum of 1851 g?m^?2 for S. bicolor var. sudanese. The average biomass was 792 g?m^?2 for B. hirta and 804 g?m^?2 for S. alba in the two field studies, respectively. The glasshouse experiment used a loamy sand field soil inoculated with a natural fine sandy loam soil that was known to contain high populations of Verticillium dahliae. Soils at both field sites belonged to the sandy loam series, and efforts were made to maintain sufficient soil moisture for plant growth. Although the interest was to determine all volatile compounds in general, only methyl sulfide and dimethyl disulfide were identified and subsequently quantified. Depending on plant species and time of sampling (one to seven days after soil incorporation), 2.7 to 346.4?μg g ^?1 plant dry weight for methyl sulfide and 0 to 283.2?μg g ^?1 plant dry weight for dimethyl disulfide were found in the glasshouse experiment. In general, high concentrations of dimethyl disulfide and methyl sulfide appeared to have reduced V. dahliae colony counts in bioassay potato stem saps in the glasshouse experiment. However, the correlation was weak (R ² ?=?0.31), but a relatively stronger correlation was obtained (R ² ?=?0.58) when excluding B. oleracea and B. rapa from the regression. Dimethyl disulfide and methyl sulfide were nearly non-detectable in the field demonstration, consequently no disease assessment was made. In the field experiment, a production of 5.2?μg g ^?1 plant dry weight for methyl sulfide and 1.2?μg g ^?1 dry weight for dimethyl disulfide was found two days after soil incorporation of S. alba. Compared to the untreated control, total Fusarium oxysporum counts in field soil were significantly lower 39 days after S. alba incorporation. However, no significant impact was found on total Pythium counts. Soil population of citrus nematode (Tylenchulus semipenetrans) in the S. alba plots was significantly reduced to similar levels found in the untreated control 112 days after S. alba incorporation. Compared to the untreated control, soil density of non plant parasitic freeliving nematodes was higher 39 days after S. alba incorporation. The study demonstrated quantifiable production of methyl sulfide and dimethyl disulfide gases from a variety of plant species in glasshouse and natural field environments. Some beneficial effects against V. dahliae, F. oxysporum, and T. semipenetrans were observed. Additional studies are needed to further elucidate these complex chemical and biological interactions.     

Carbon-13 and proton nuclear magnetic resonance studies on methyl aldofuranosides and their O-alkyl derivatives

The effect of O-alkylation on the carbon-13 n.m.r. spectra of methyl pentofuranosides has been determined. O-Alkylation of an OH group displaced the signal of the appended.¹³C nucleus downfield, whereas the adjacent ¹³C nuclei were, in most instances, shifted upfield to a smaller extent. The effect of O-methylation was appreciably larger than O-isopropylation or O-glycosylation, but either O-methyl or O-isopropyl derivatives may be used as models for interpreting the spectra of furanoid oligo- and poly-saccharides, including the galactomannan of Penicillium charlesii. The signal displacements are, to a large extent, comparable to those observed in the conformationally more stable mannopyranose series, so that they are insensitive to effects of steric distortion and population (or both). These effects occurred on 3-O-alkylation of methyl pentofuranosides, as appreciable changes in J_1,2 values in their p.m.r. spectra were observed.     

Effects of methyl mercury at different dose regimes on eggshell formation and some biochemical characteristics of the eggshell gland mucosa of the domestic fowl

Eggshell formation and egg production in domestic fowl were studied following the administration of methyl mercury (two dose regimes: 5 mg daily for 6 consecutive days and 1 mg daily for 50 consecutive days). A daily oral dose of 5 mg of methyl mercury for 6 consecutive days induced significant eggshell thinning and deformation and inhibited egg production. Uptake of ⁴⁵Ca and synthesis of prostaglandins by a homogenate of eggshell gland mucosa from methyl-mercury-treated birds were significantly reduced, as was the calcium content of blood plasma. A daily oral dose of 1 mg of methyl mercury administered for 50 consecutive days also induced eggshell deformation and thinning and reduced egg production. This dose did not, however, have significant effects on the following: ⁴⁵Ca uptake and prostaglandin synthesis by a homogenate of the eggshell gland mucosa; ⁴⁵Ca uptake by a homogenate of duodenal mucosa; the Ca content of the blood plasma, shell gland mucosa or shell gland lumen; the HCO₃⁻ content of the shell gland lumen or the specific gravity of tibia. Methyl mercury added in vitro to a homogenate of eggshell gland mucosa significantly stimulated the synthesis of prostaglandins PGF_2α and PGE₂. Addition of mercury chloride to the same type of preparation stimulated the synthesis of PGF_2α at the expense of thromboxane (T × B₂) synthesis. Administration of 5 mg methyl mercury for 6 consecutive days seemed to reduce the availability of calcium for eggshell formation. This effect could have been due to a direct inhibitory effect of methyl mercury on calcium uptake from the gastrointestinal tract and/or to mobilization of medullary bone. The administration of 1 mg methyl mercury for 50 consecutive days probably induced the reproductive effects by another mechanism. The effects of methyl mercury on avian eggshell formation are quite different from the effects p,p′-DDE exerts on that process.     

Free fatty acids from the pasture grass Brachiaria humidicola and one of their methyl esters as inhibitors of nitrification

The tropical pasture grass, Brachiaria humidicola (Rendle) Schweick, produces nitrification inhibitory compounds (termed biological nitrification inhibitors or BNIs) in its shoot and root tissues and releases BNIs from its roots. In the present study, two BNI compounds were isolated and identified from the shoot tissue of B. humidicola using activity-guided fractionation. The recombinant Nitrosomonas europaea containing luxAB genes derived from the bioluminescent marine gram-negative bacterium Vibrio harveyi, were used to determine BNI activity. The BNI compounds in the shoot tissue were identified as linoleic acid (LA) and linolenic acid (LN) using authentic-chemicals obtained from ©Sigma (ED₈₀ 16.0 μg ml^?1 for both LA and LN) for verification. None of the other tested free fatty acids namely stearic acid, oleic acid, arachidonic acid, and cis-vaccenic acid showed any inhibitory effect on nitrification. Among the fatty acid methyl esters (FAME) evaluated [methyl oleate, methyl linoleate (LA-ME) and methyl linoleneate (LN-ME)], only LA-ME showed an inhibitory effect (ED₈₀ 8.0 μg ml^?1). The inhibitory effect of LA, LN and LA-ME in the soil was stable for 120 days at 20°C. Soil treated with LA, LN and LA-ME showed a very low accumulation of NO₃ ^? and the maintenance of soil inorganic N in the NH₄ ⁺ form. The inhibitory effect of LA-ME on soil nitrification was greater than that of LA or LN. In addition to BNI activity, both LA and LA-ME showed a suppressive effect on urea hydrolysis in soil. Both LA and LN blocked the AMO (ammonia monooxygenase) and HAO (hydroxylamino oxidoreductase) enzymatic pathways in Nitrosomonas. Since LA and LN can be produced from vegetable oils such as soybean, flax or sunflower, they have the potential for use as nitrification inhibitors in production agriculture.