Fatty acids. part 43. The preparation and properties of some methyl dimercaptostearates and of some related methyl epithio-and epidithiostearates

The methods described for converting mesyloxy esters to mercapto esters (see previous paper) have been applied to appropriate dimesyloxy derivatives as a route to the 9,10; 10,12 and 9,12-dimercapostearates. Some reactions furnish epithio and epidithio esters in place of mercapto esters. The dithiols are converted to acetyl, trifluoroacetyl, and isopropylidene derivatives. The products are identified mainly by infrared, nuclear magnetic resonance, and mass spectroscopic procedures.     

Biosynthesis of acyclic methyl branched polyunsaturated hydrocarbons inPseudomonas maltophilia

Summary The hydrocarbon composition ofPseudomonas maltophilia was determined by gas chromatography-mass spectrometry. Mono-, di- and tri-unsaturated alkenes were identified with a predominance of polyunsaturated components. The carbon chains of the alkenes contained single methyl branches iniso andanteiso position and double methyl branches in theiso-iso andanteiso-anteiso configurations. The composition of the hydrocarbons from cells grown in synthetic media enriched with amino acids or volatile fatty acids demonstrated that the probable precursors incorporated into individual hydrocarbons were branched and normal fatty acid chains in the range from C₃ to C₁₆. The probable fatty acid precursors which were connected together to form the major triunsaturated hydrocarbon chains were two monounsaturated chains, whereas the major liunsaturated chains resulted from condensation of saturated and monounsaturated chains. The probable precursors for the major monounsaturated hydrocarbons were C₁₄ (C₁₅) and C₁₆ (C₁₅) fatty acids. The accumulation of hydrocarbons was not detected until the cells were in the late exponential phase of growth; the maximal levels were reached at the mid-stationary phase of growth.     

Epoxidation of fatty acids,fatty methyl esters,and alkenes by immobilized oat seed peroxygenase

Fatty epoxides are used as plasticizers and plastic stabilizers and are intermediates for the production of other chemical substances. The currently used industrial procedure for fatty epoxide synthesis requires a strong acid catalyst which can cause oxirane ring opening and side product formation. To find a replacement for the acid catalyst, we have been conducting research on a peroxygenase enzyme from oat (Avena sativa) seeds and have devised a method for immobilization of this enzyme using a hydrophobic membrane support. In this study, fatty acids and fatty methyl esters commonly encountered in commercial vegetable oils were tested as substrates for immobilized peroxygenase, and the epoxide products were characterized. The epoxidation time course of linoleic acid showed two distinct phases with nearly complete conversion to monoepoxide before diepoxide was produced. The diepoxide formed from linolenic acid was found to be 9,10-15,16-diepoxy-12-octadecenoic acid, and only a trace of triepoxide was obtained. Additionally it was discovered that acyclic alkenes with internal double bonds, a cyclic alkene, and an alkene with an aromatic substituent were substrates of peroxygenase. However, alkenes with terminal unsaturation were unreactive. With every substrate examined, oat seed peroxygenase exhibited specificity for epoxidation, producing no other products, and oxirane ring opening did not occur.     

The occurrence of monounsaturated n-C21 and polyunsaturated C25 sedimentary hydrocarbons in the lipids of Antarctic marine organisms

Antarctic zooplankton have been found to be a potential source of sedimentary hydrocarbons. Monounsaturated C₂₁ n-alkenes and highly branched polyunsaturated C₂₅ n-alkenes were analysed in the aliphatic fraction of the lipids of Antarctic pelagic and inshore marine organisms. Cluster analysis of the species-based data set produced four main groups: phytoplankton, epipelagic herbivores, epipelagic carnivores and mesopelagic omnivores. The detailed pattern of alkenes exhibited differences within the groups and also with tissue type (krill, Euphausia superba). The origin of alkenes in Antarctic biota appeared to be either synthesis de novo or due to the condensation of smaller molecules. Formation of alkenes by the decarboxylation of fatty acids was not consistent with the hydrocarbon and fatty acid composition of Antarctic zooplankton. There was no evidence for direct assimilation of C₂₁ and C₂₅ alkenes by zooplankton or higher predators from their diet. Zooplankton C₂₅ alkenes are probably transported unaltered directly to the sediment as detritus or via predators in faecal material. Sedimentary C₂₅ alkenes are proposed as biomarkers of recent zooplankton activity in the water column.     

Applications of chromic acid-celite columns to lipid analysis. Location of double bond position in submicro- and microgram amounts of methyl octadecenoates.

A procedure for locating the double bond position in a series of methyl octadecenoates is detailed. Submicrogram or microgram amounts of substrate dissolved in CS₂ are brought in contact with a very small column of chromic acid on Celite, and the oxidation products (car?ylic acids) are eluted, converted to methyl esters, and resolved by gas-liquid chromatography. Beside the acids resulting from scission of the double bond, acids containing one less carbon atom arise from oxidation of the allylic carbons on both sides of the double bond so that pairs of peaks appear on the chromatogram. All positions from Δ3 to Δ17 were located successfully. The Δ2 position failed to oxidize.     

Hydrocarbons obtained by pyrolysis of some precambrian rocks of minnesota

Drill core samples of 42 Precambrian sedimentary, igneous, and metamorphic rocks were analyzed by heating under partial vacuum at 100°C and at 400°C to release hydrocarbons and other volatile products.The core samples yielded methane in amounts ranging from traces to 3 microliters per gram, but averaged much less. By way of comparison, samples of Middle Devonian Marcellus black shale, from Pennsylvania, yielded methane in amounts up to 7ul/g.Other straight chain hydrocarbons up to C₁₁ were found in the volatile products, especially those obtained at 400°C, and benzene was a common product, also mainly in the 400°C experiments. Carbon dioxide and nitrogen appear to form a large part of the nonhydrocarbon volatiles in at least some of the samples.Spectral data indicate that the straight chain pyrolysis products of the Precambrian rocks are mainly alkenes, whereas those of the Devonian rocks, referred to above, are a mixture of alkanes and alkenes. Alkanes were however, obtained from several algae-bearing Middle Precambrian argillites. Available evidence indicates, although not conclusively, that the alkenes were contained in the rock rather than being produced from alkanes during pyrolysis.The writers believe that surface contamination in most of the drill cores was minimal owing to the low permeability of the rocks studied, and that contamination by drilling was also minimal.There is a reasonable possibility that the volatiles, if not formed from kerogen residues by the pyrolysis experiments, are in part juvenile igneous gases or are substances that were distilled out of the deeperlying rocks during intervals of folding and metamorphism, and subsequently accumulated at higher levels.     

Diethyl‐Ether Flower Washings of Dianthus cruentus Griseb. (Caryophyllaceae): Derivatization Reactions Leading to the Identification of New Wax Constituents

Some carnation species (Dianthus spp., Caryophyllaceae) exhibit a strong resistance to drought stress that is connected with the increased surface wax formation. Wax composition is unknown for the majority of Dianthus spp. Herein, mass spectral and gas chromatographic data, in combination with synthesis and chemical transformations (transesterification and synthesis of dimethyl disulfide adducts), enabled the identification of 151 constituents of diethyl‐ether washings of fresh flowers of Dianthus cruentus Griseb . from Serbia. The flower wax contained, along with the dominant ubiquitous long‐chain n‐alkanes, homologous series of n‐ and branched (iso‐ and anteiso‐) long‐chain hexyl alkanoates/alkenoates and alkyl/alkenyl benzoates. The branching position in the mentioned hexyl esters was probed by synthesis of esters of three isomeric hexanols that were spectrally characterized (¹H‐ and ¹³C‐NMR, IR, MS). The washings also contained long‐chain (Z)‐ and (E)‐alkenes (C₂₃?C₃₅) with several different double bond regiochemistries. Fifty‐five of these constituents (eight hexyl esters, two benzoates, and forty‐five alkenes) were detected for the first time in Plantae, while ten of these represent completely new compounds. The rare occurrence of these wax constituents makes them possible chemotaxonomic markers of this particular Dianthus sp.     

Properties and metabolism of 2alkylalkanoates. 3. absorption of methyl and ethyl 2-methylpalmitate

The recovery from rat and rabbit tissues of fed methyl-(14)C and ethyl-2-(14)C 2-methylpalmitate with unaltered specific activity has demonstrated the existence of mechanisms for the absorption and deposition of both methyl and ethyl esters of fatty acids, at least for 2-methylpalmitate. In thoracic duct-cannulated rats, approximately 9% of the fed compounds was recovered from the lymph during the first 24 hr, the rate of recovery reaching a maximum between 6 and 8 hr. In the rabbit, the fed, unaltered esters in plasma were transported principally by means of the low density lipoproteins. Only trace amounts of the unaltered esters were subsequently detected in the blood and tissue lipids after feeding, however, even during the period of maximal absorption; moreover, in contrast to at least one report by others, further analyses for methyl or ethyl esters of other fatty acids has shown that such esters of short-chain alcohols constitute no more than a trace amount (0.004-1.03%) of the lipids extracted from a wide variety of mammalian tissues. The possibility remains that even these trace amounts of esters arose as artifacts of autolysis, extraction, or assay.     

Simultaneous capillary gas chromatographic profiling of medium- and long-chain fatty acid methyl esters with split injection: Correction for injection-related discrimination by the ‘bracketing’ method

Split injection-related discrimination can be a source of inaccuracy and imprecision in quantitative capillary gas chromatographic profiling methods for compounds with relatively big differences in boiling points, such as the methyl esters of medium- and long-chain fatty acids prepared from biological materials. We systematically investigated a standard containing equal masses of saturated fatty acid methyl esters, with chain lengths from C₅ to C₂₆, under different injection conditions, including injection temperature, sample volume and split ratio. Day-to-day performance was studied under one set of conditions. Normalized peak areas, reciprocal response factors, using either C₁₇ or C₂₃ as an internal standard, and ‘bracketed’ reciprocal response factors (peak area of each analyte divided by half the sum of the peak areas of two adjacent esters were calculated. In all experiments the bracketed reciprocal response factors were found to be closest to unity with the lowest coefficients of variation.     

Biological activity of some synthetic gibberellin glucosyl esters

Four gibberellin (GA₁, GA₃, GA₄ and GA₃₇) glucosyl esters were synthesized and found to be as active as their respective free acids in the rice seedling bioassay. The rapid hydrolysis of the glucosyl esters in rice seedlings was demonstrated by feeding experiments with glucosyl esters of [³H]GA₁ and [³H]GA₄.     

Isolation and characterization of fatty acid methyl ester (FAME)‐producing Streptomyces sp. S161 from sheep (Ovis aries) faeces

An actinomycete producing oil‐like mixtures was isolated and characterized. The strain was isolated from sheep faeces and identified as Streptomyces sp. S161 based on 16S rRNA gene sequence analysis. The strain showed cellulase and xylanase activities. The ¹H nuclear magnetic resonance (NMR) spectra of the mixtures showed that the mixtures were composed of fatty acid methyl esters (52·5), triglycerides (13·7) and monoglycerides (9·1) (mol.%). Based on the gas chromatography–mass spectrometry (GC‐MS) analysis, the fatty acid methyl esters were mainly composed of C14‐C16 long‐chain fatty acids. The results indicated that Streptomyces sp. S161 could produce fatty acid methyl esters (FAME) directly from starch. To our knowledge, this is the first isolated strain that can produce biodiesel (FAME) directly from starch.

Significance and Impact of the Study

Nowadays, production of biodiesel is based on plant oils, animal fats, algal oils and microbial oils. Lipid mostly consists of triacylglycerols (TAG), and conversion of these lipids into fatty acid short‐chain alcohol esters (methanol or ethanol) is the final step in biodiesel production. In this study, an oil‐producing Streptomyces strain was isolated from sheep faeces. The oil was composed of C₁₄‐C₁₆ long‐chain fatty acid methyl esters, triglycerides and monoglycerides. This is the first isolated strain‐producing biodiesel (FAME) directly from starch. Due to showing cellulase and xylanase activities, the strain would be helpful for converting renewable lignocellulose into biodiesel directly.     

Papain-catalysed hydrolysis of some hippuric esters. A new mechanism for papain-catalysed hydrolyses

1. The Michaelis–Menten parameters for the papain-catalysed hydrolysis of a number of alkyl, aryl and alkyl-thiol esters of hippuric acid have been determined. 2. For all the aryl esters and most of the alkyl esters studied, the catalytic constant, k₀, is 2–3sec.⁻¹ and most probably represents deacylation of the common intermediate, hippuryl-papain. 3. Two alkyl esters and hippurylamide, however, have catalytic rate constants, k₀, less than 2–3sec.⁻¹. It is possible to interpret all the available kinetic data in terms of a three-step mechanism in which an enzyme–substrate complex is first formed, followed by acylation of the enzyme through an essential thiol group, followed by deacylation of the acyl-enzyme. 4. The logarithm of the ratio of the Michaelis–Menten parameters, which reflect the acylation rate constant, for four aryl esters of hippuric acid studied give a linear Hammett plot against the substituent constant, σ. Arguments are presented that indicate acid as well as nucleophilic catalysis in the acylation process and that the most likely proton donor is an imidazolium ion. 5. It is suggested that this imidazolium ion is part of the same histidine residue that has been tentatively implicated in the deacylation process (Lowe & Williams, 1965b). 6. A new mechanism is proposed for the papain-catalysed hydrolysis of N-acyl-α-amino acid derivatives.     

Oxidation of Gaseous and Volatile Hydrocarbons by Selected Alkene-Utilizing Bacteria

Eleven strains of alkene-utilizing bacteria belonging to the genera Mycobacterium, Nocardia, and Xanthobacter were tested for their ability to grow with C₁ to C₆ alkanes, C₂ to C₆ alkenes, alkadienes, and monoterpenes furnished individually as sole sources of carbon and energy in a mineral salts medium. A limited number of alkenes and alkanes supported growth of the bacteria; some bacteria were unable to grow on any of the saturated hydrocarbons tested. Monoterpenes were frequently used as carbon and energy sources by alkene-utilizing bacteria belonging to the genera Mycobacterium and Nocardia. Washed cell suspensions of alkene-grown bacteria attacked the whole range of alkenes tested, whereas only three strains were able to oxidize alkanes as well. The alkenes tested were oxidized either to water and carbon dioxide or to epoxyalkanes. Few epoxides accumulated in stoichiometric amounts from the corresponding alkenes, because most epoxides formed were further converted to other compounds like alkanediols.     

Microbial Oxidation of Hydrocarbons: Properties of a Soluble Methane Monooxygenase from a Facultative Methane-Utilizing Organism, Methylobacterium sp. Strain CRL-26

Methylobacterium sp. strain CRL-26 grown in a fermentor contained methane monooxygenase activity in soluble fractions. Soluble methane monooxygenase catalyzed the epoxidation/hydroxylation of a variety of hydrocarbons, including terminal alkenes, internal alkenes, substituted alkenes, branched-chain alkenes, alkanes (C₁ to C₈), substituted alkanes, branched-chain alkanes, carbon monoxide, ethers, and cyclic and aromatic compounds. The optimum pH and temperature for the epoxidation of propylene by soluble methane monooxygenase were found to be 7.0 and 40°C, respectively. Among various compounds tested, only NADH₂ or NADPH₂ could act as an electron donor. Formate and NAD⁺ (in the presence of formate dehydrogenase contained in the soluble fraction) or 2-butanol in the presence of NAD⁺ and secondary alcohol dehydrogenase generated the NADH₂ required for the methane monooxygenase. Epoxidation of propylene catalyzed by methane monooxygenase was not inhibited by a range of potential inhibitors, including metal-chelating compounds and potassium cyanide. Sulfhydryl agents and acriflavin inhibited monooxygenase activity. Soluble methane monooxygenase was resolved into three components by ion-exchange chromatography. All three compounds are required for the epoxidation and hydroxylation reactions.     

Carbonic anhydrase inhibitors. Selective inhibition of human tumor-associated isozymes IX and XII and cytosolic isozymes I and II with some substituted-2-mercapto-benzenesulfonamides

A series of 2-mercapto-substituted-benzenesulfonamides has been prepared by a unique two-step procedure starting from the corresponding 2-chloro-substituted benzenesulfonamides. Compounds bearing an unsubstituted mercapto group and the corresponding S-benzoyl derivatives were investigated as inhibitors of four isoforms of the zinc enzyme carbonic anhydrase (CA, EC 4.2.1.1), i.e., the cytosolic, ubiquitous isozymes CA I and II, as well as the transmembrane, tumor associated isozymes CA IX and XII. These derivatives were medium potency hCA I inhibitors (K_Is in the range of 1.5–5.7 μM), two derivatives were strong hCA II inhibitors (K_Is in the range of 15–16 nM), whereas the others showed weak activity. These compounds inhibited hCA IX with inhibition constants in the range 160–1950 nM and hCA XII with inhibition constants in the range 1.2–413 nM. Some of these derivatives showed a certain degree of selectivity for inhibition of the tumor-associated over the cytosolic isoforms, being thus interesting leads for the development of potentially novel applications in the management of hypoxic tumors which overexpress CA IX and XII.     

大花金挖耳不同部位挥发油化学成分比较分析

采用GC-MS技术,对利用水蒸气蒸馏法从大花金挖耳根、茎、叶、花及花托等5个部位获得的挥发油组分进行了分析.从大花金挖耳5种挥发油中共鉴定出99种成分,从根、茎、叶、花及花托分别鉴定出了37、34、47、29和40种化合物,已鉴定的组分分别占相应挥发油的85.86%、84.98%、83.06%、84.58%和89.06%.5个部位挥发油化学组成差异较大,根挥发油主要成分为倍半萜类和酯类;茎挥发油主要为倍半萜和烯类(包括烯醇、烯酸、烯酮);叶和花挥发油主要为烯类(包括烯醇、烯酸、烯酮)和酯类;花托挥发油主要为倍半萜类和二萜类.     

Facile syntheses of <Emphasis Type="SmallCaps">l</Emphasis>-β-haloalanine derivatives from <Emphasis Type="SmallCaps">l</Emphasis>-cysteine or <Emphasis Type="SmallCaps">l</Emphasis>-cystine

l-β-Haloalanines are physiologically active unnatural amino acids and they are useful intermediates for the synthesis of natural and unnatural amino acids, S-linked glycopeptides, and lanthionines. In general l-β-haloalanines were prepared predominantly from l-serine via functional group transformation. Here we reported an alternative approach for the preparation of l-β-haloalanines via halogenation of protected l-cysteine esters which was obtained from l-cysteine or l-cystine, respectively. The mercapto group of protected l-cysteine esters was efficiently transformed to halo groups by triphenylphosphine/N-halosuccinimides. It has been proved to be a versatile desulfurization strategy via this functional group transformation.     

Straight phase separation of prostaglandin methyl esters on lipophilic gels

A series of straight phase gel chromatography systems have been developed for the separation of prostaglandin methyl esters. Using the methyl esters of prostaglandins B₂, E₂, F₂α and F₂β, the basic relationships between elution volume and the polarities of the gel, the solvent system (heptane-chloroform mixtures), and the prostaglandin have been determined. The separation of prostaglandin methyl esters with slight differences in structure has been demonstrated. Examples include oxo and hydroxy prostaglandins, hydroxy epimers, double bond isomers, prostaglandins of varying α- and ω-chain length, and 1- and 2- (5,6 cis double bond) series prostaglandins. In view of the general advantages of liquid-gel chromatography, it is suggested that these systems may be useful for isolation and purification in a number of areas in the prostaglandin field.     

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.     

Microbial‐based motor fuels: science and technology

The production of biofuels via microbial biotechnology is a very active field of research. A range of fuel molecule types are currently under consideration: alcohols, ethers, esters, isoprenes, alkenes and alkanes. At the present, the major alcohol biofuel is ethanol. The ethanol fermentation is an old technology. Ongoing efforts aim to increase yield and energy efficiency of ethanol production from biomass. n‐Butanol, another microbial fermentation product, is potentially superior to ethanol as a fuel but suffers from low yield and unwanted side‐products currently. In general, biodiesel fuels consist of fatty acid methyl esters in which the carbon derives from plants, not microbes. A new biodiesel product, called microdiesel, can be generated in engineered bacterial cells that condense ethanol with fatty acids. Perhaps the best fuel type to generate from biomass would be biohydrocarbons. Microbes are known to produce hydrocarbons such as isoprenes, long‐chain alkenes and alkanes. The biochemical mechanisms of microbial hydrocarbon biosynthesis are currently under study. Hydrocarbons and minimally oxygenated molecules may also be produced by hybrid chemical and biological processes. A broad interest in novel fuel molecules is also driving the development of new bioinformatics tools to facilitate biofuels research.