Polyhydroxy Fatty Acids Research Articles

7,10-Dihydroxy-8(E)-octadecenoic Acid Displays a Fungicidal Activity against Malassezia furfur.

Microbial conversion of some natural unsaturated fatty acids can produce polyhydroxy fatty acids, giving them new properties, such as higher viscosity and reactivity. Pseudomonas aeruginosa has been intensively studied to produce a novel 7,10-dihydroxy-8(E)-octadecenoic acid (DOD) from oleic acid and natural vegetable oils containing oleic acid. Recently, the antibacterial activities of DOD against food-borne pathogenic bacteria were reported; however, the action of such antibacterial properties against eucaryotic cells remains poorly known. In this study, we determined the antifungal activities of DOD against Malassezia furfur KCCM 12679 quantitatively and qualitatively. The antifungal activity of DOD against M. furfur KCCM 12679 was approximately five times higher than that of ketoconazole, a commercial antifungal agent. The MIC 90 value of DOD against M. furfur KCCM 12679 was 50 µg/mL. In addition, we confirmed that the antifungal property of DOD was exerted through fungicidal activity.

Hydrolysis of polyhydroxy polyunsaturated fatty acid-glycerophosphocholines by Group IIA, V, and X secretory phospholipases A2.

It is widely accepted that unesterified polyunsaturated ω-6 and ω-3 fatty acids (PUFA) are converted through various lipoxygenases, cyclooxygenases, and cytochrome P450 enzymes to a range of oxygenated derivatives (oxylipins), among which the polyhydroxides of unesterified PUFA have recently been recognized as cell signaling molecules with anti-inflammatory and pro-resolving properties, known as specialized pro-resolving mediators (SPMs). This study investigates the mono-, di-, and trihydroxy 16:0/PUFA-GPCs, and the corresponding 16:0/SPM-GPC, in plasma lipoproteins. We describe the isolation and identification of mono-, di-, and trihydroxy AA, EPA, and DHA-GPC in plasma LDL, HDL, HDL3, and acute phase HDL using normal phase LC/ESI-MS, as previously reported. The lipoproteins contained variable amounts of the polyhydroxy-PUFA-GPC (0-10nmol/mg protein), likely the product of lipid peroxidation and the action of various lipoxygenases and cytochrome P450 enzymes on both free fatty acids and the parent GPCs. Polyhydroxy-PUFA-GPC was hydrolyzed to variable extent (20%-80%) by the different secretory phospholipases A2 (sPLA2 s), with Group IIA sPLA2 showing the lowest and Group X sPLA2 the highest activity. Surprisingly, the trihydroxy-16:0/PUFA-GPC of APHDL was largely absent, while large amounts of unidentified material had migrated in the free fatty acid elution area. The free fatty acid mass spectra were consistent with that anticipated for branched chain polyhydroxy fatty acids. There was general agreement between the masses determined by LC/ESI-MS for the polyhydroxy PUFA-GPC and the masses calculated for the GPC equivalents of resolvins, protectins, and maresins using the fatty acid structures reported in the literature.

Valorization of Agro-Industrial Wastes by Ultrasound-Assisted Extraction as a Source of Proteins, Antioxidants and Cutin: A Cascade Approach.

The use of agro-industrial wastes to obtain compounds with a high added-value is increasing in the last few years in accordance with the circular economy concept. In this work, a cascade extraction approach was developed based on ultrasound-assisted extraction (UAE) for tomato, watermelon, and apple peel wastes. The protein and antioxidant compounds were obtained during the first extraction step (NaOH 3 wt.%, 98.6 W, 100% amplitude, 6.48 W/cm2, 6 min). The watermelon peels (WP) showed higher proteins and total phenolic contents (857 ± 1 mg BSA/g extract and 107.2 ± 0.2 mg GAE/100 g dm, respectively), whereas the highest antioxidant activity was obtained for apple peels (1559 ± 20 µmol TE/100 g dm, 1767 ± 5 µmol TE/100 g dm, and 902 ± 16 µmol TE/100 g dm for ABTS, FRAP and DPPH assays, respectively). The remaining residue obtained from the first extraction was subsequently extracted to obtain cutin (ethanol 40 wt.%, 58 W, 100% amplitude, 2 W/cm2, 17 min, 1/80 g/mL, pH 2.5). The morphological studies confirmed the great efficiency of UAE in damaging the vegetal cell walls. WP showed a higher non-hydrolysable cutin content (55 wt.% of the initial cutin). A different monomers’ profile was obtained for the cutin composition by GC-MS, with the cutin from tomato and apple peels being rich in polyhydroxy fatty acids whereas the cutin extracted from WP was mainly based on unsaturated fatty acids. All of the cutin samples showed an initial degradation temperature higher than 200 °C, presenting an excellent thermal stability. The strategy followed in this work has proved to be an effective valorization methodology with a high scaling-up potential for applications in the food, pharmaceutical, nutraceutical, cosmetics and biopolymer sectors.

Boron Isotope Tag-Assisted Ultrahigh-Performance Liquid Chromatography Coupled with High-Resolution Mass Spectrometry for Discovery and Annotation of cis-Diol-Containing Metabolites.

cis-Diol-containing metabolites are widely distributed in living organisms, and they participate in the regulation of various important biological activities. The profiling of cis-diol-containing metabolites could help us in fully understanding their functions. In this work, based on the characteristic isotope pattern of boron, we employed a boronic acid reagent as the isotope tag to establish a sensitive and selective liquid chromatography-high-resolution mass spectrometry method for the screening and annotation of cis-diol-containing metabolites in biological samples. Boronic acid reagent 2-methyl-4-phenylaminomethylphenylboronic acid was used to label the cis-diol-containing metabolites in biological samples to improve the selectivity and MS sensitivity of cis-diol-containing metabolites. Based on the characteristic 0.996 Da mass difference of precursor ions and the peak intensity ratio of 1:4 originating from 10B and 11B natural isotopes, the potential cis-diol-containing metabolites were rapidly screened from biological samples. Potential cis-diol-containing metabolites were annotated by database searching and analysis of fragmentation patterns obtained by multistage MS (MSn) via collision-induced dissociation. Importantly, the cis-diol position could be readily resolved by the MS3 spectrum. With this method, a total of 45 cis-diol-containing metabolites were discovered in rice, including monoglycerides, polyhydroxy fatty acids, fatty alcohols, and so forth. Furthermore, the established method showed superiority in avoiding false-positive results in profiling cis-diol-containing metabolites.

Drought-Responsive ZmFDL1/MYB94 Regulates Cuticle Biosynthesis and Cuticle-Dependent Leaf Permeability.

In all land plants, the outer surface of aerial parts is covered by the cuticle, a complex lipid layer that constitutes a barrier against damage caused by environmental factors and provides protection against nonstomatal water loss. We show in this study that both cuticle deposition and cuticle-dependent leaf permeability during the juvenile phase of plant development are controlled by the maize (Zea mays) transcription factor ZmFUSED LEAVES 1 (FDL1)/MYB94. Biochemical analysis showed altered cutin and wax biosynthesis and deposition in fdl1-1 mutant seedlings at the coleoptile stage. Among cutin compounds, ω-hydroxy fatty acids and polyhydroxy-fatty acids were specifically affected, while the reduction of epicuticular waxes was mainly observed in primary long chain alcohols and, to a minor extent, in long-chain wax esters. Transcriptome analysis allowed the identification of candidate genes involved in lipid metabolism and the assembly of a proposed pathway for cuticle biosynthesis in maize. Lack of ZmFDL1/MYB94 affects the expression of genes located in different modules of the pathway, and we highlighted the correspondence between gene transcriptional variations and biochemical defects. We observed a decrease in cuticle-dependent leaf permeability in maize seedlings exposed to drought as well as abscisic acid treatment, which implies coordinated changes in the transcript levels of ZmFDL1/MYB94 and associated genes. Overall, our results suggest that the response to water stress implies the activation of wax biosynthesis and the involvement of both ZmFDL1/MYB94 and abscisic acid regulatory pathways.

Cutin from watermelon peels: A novel inducer for cutinase production and its physicochemical characterization

Cutin a polymeric biological macromolecule made up of esterified polyacids which acts as an inducer of cutinase. The present work was based on screening and characterization of newer cutin source from the agro industrial wastes as inducers of cutinase production by Fusarium oxysporum MTCC 2480. Cutin isolated from peels of multi green colored watermelon (WMC) yielded 6.77U/mL as compared to that 9.64U/mL of cutinase using apple cutin. The FTIR and 13C CP-MAS solid state NMR studies indicated the nature of WMC to be an aliphatic polyester of polyhydroxy fatty acids. A further FTIR spectroscopic study on dewaxed cutin confirmed that the ester linkages in WMC were completely hydrolyzed during submerged fermentation. GC–MS data clearly indicated the critical structural feature of WMC viz. hydroxyl groups at ω-position and middle of the fatty acid chain. The amorphous nature of WMC was confirmed by XRD. DSC of WMC showed two endothermic transition peaks, one broad appearing at 30–60°C and other at 145°C. Thermogravimetric analysis of WMC suggested it to be thermally stable up to 200°C. This study warrants further work on using WMC as a substrate for fermentative production of cutinase.

Pectin-lipid self-assembly: influence on the formation of polyhydroxy fatty acids nanoparticles.

Nanoparticles, named cutinsomes, have been prepared from aleuritic (9,10,16-trihidroxipalmitic) acid and tomato fruit cutin monomers (a mixture of mainly 9(10),16-dihydroxypalmitic acid (85%, w/w) and 16-hydroxyhexadecanoic acid (7.5%, w/w)) with pectin in aqueous solution. The process of formation of the nanoparticles of aleuritic acid plus pectin has been monitored by UV-Vis spectrophotometry, while their chemical and morphological characterization was analyzed by ATR-FTIR, TEM, and non-contact AFM. The structure of these nanoparticles can be described as a lipid core with a pectin shell. Pectin facilitated the formation of nanoparticles, by inducing their aggregation in branched chains and favoring the condensation between lipid monomers. Also, pectin determined the self-assembly of cutinsomes on highly ordered pyrolytic graphite (HOPG) surfaces, causing their opening and forming interconnected structures. In the case of cutin monomers, the nanoparticles are fused, and the condensation of the hydroxy fatty acids is strongly affected by the presence of the polysaccharide. The interaction of pectin with polyhydroxylated fatty acids could be related to an initial step in the formation of the plant biopolyester cutin.

Polyols and polyurethanes prepared from epoxidized soybean oil ring‐opened by polyhydroxy fatty acids with varying OH numbers

AbstractBio‐based polyols from epoxidized soybean oil and different fatty acids were successfully prepared using a solvent‐free method in order to investigate the effect of the polyols' OH numbers on the thermal and mechanical properties of the polyurethanes prepared using them. Epoxidized soybean oil/epoxidized linseed oil was ring‐opened by methanol/glycol followed by saponification to prepare polyhydroxy fatty acids. These fatty acids and epoxidized soybean oil were then used for further solvent‐free ring‐opening reactions with DBU as catalyst, which facilitated the carboxylic ring‐opening. Gel permeation chromatography revealed that a molar ratio of carboxylic acid from polyhydroxy fatty aicds and epoxy group of 0.5 : 1 resulted in optimized polyols containing the smallest amounts of residual starting materials. The obtained polyols had varying OH numbers and the acquired polyurethane films were comprehensively characterized. With increasing OH number of the polyols the PUs displayed an increase in crosslinking density, glass transition temperature (Tg), tensile strength and Young's modulus, and a decrease in elongation and toughness. This work provides Supporting Information on the effect of OH number of polyols obtained via a solvent‐free ring‐opening method on the mechanical and thermal properties of polyurethanes, of particular interest when designing PU products for specific purposes. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015, 132, 41213.

Cutinsomes and lipotubuloids appear to participate in cuticle formation in Ornithogalum umbellatum ovary epidermis: EM–immunogold research

The outer wall of Ornithogalum umbellatum ovary and the fruit epidermis are covered with a thick cuticle and contain lipotubuloids incorporating 3H-palmitic acid. This was earlier evidenced by selective autoradiographic labelling of lipotubuloids. After post-incubation in a non-radioactive medium, some marked particles insoluble in organic solvents (similar to cutin matrix) moved to the cuticular layer. Hence, it was hypothesised that lipotubuloids participated in cuticle synthesis. It was previously suggested that cutinsomes, nanoparticles containing polyhydroxy fatty acids, formed the cuticle. Thus, identification of the cutinsomes in O. umbellatum ovary epidermal cells, including lipotubuloids, was undertaken in order to verify the idea of lipotubuloid participation in cuticle synthesis in this species. Electron microscopy and immunogold method with the antibodies recognizing cutinsomes were used to identify these structures. They were mostly found in the outer cell wall, the cuticular layer and the cuticle proper. A lower but still significant degree of labelling was also observed in lipotubuloids, cytoplasm and near plasmalemma of epidermal cells. It seems that cutinsomes are formed in lipotubuloids and then they leave them and move towards the cuticle in epidermal cells of O. umbellatum ovary. Thus, we suggest that (1) cutinsomes could take part in the synthesis of cuticle components also in plant species other than tomato, (2) the lipotubuloids are the cytoplasmic domains connected with cuticle formation and (3) this process proceeds via cutinsomes.

Identification of TAG and DAG and their FA Constituents in Lesquerella (Physaria fendleri) Oil by HPLC and MS

AbstractCastor oil has many industrial uses because of its high content (90 %) of the hydroxy fatty acid, ricinoleic acid (OH1218:19). Lesquerella oil containing lesquerolic acid (Ls, OH1420:111) is potentially useful in industry. Ten molecular species of diacylglycerols and 74 molecular species of triacylglycerols in lesquerella (Physaria fendleri) oil were identified by electrospray ionization mass spectrometry as lithium adducts of acylglycerols in the HPLC fractions of lesquerella oil. Among them were: LsLsO, LsLsLn, LsLsL, LsLn–OH20:2, LsO–OH20:2 and LsL–OH20:2. The structures of the four new hydroxy fatty acid constituents of acylglycerols were proposed by the MS of the lithium adducts of fatty acids as (comparing to those in castor oil): OH1218:29,14 (OH1218:29,13 in castor oil), OH1218:39,14,16 (OH18:3 not detected in castor oil), diOH12,1318:29,14 (diOH11,1218:29,13 in castor oil) and diOH13,1420:111 (diOH20:1 not detected in castor oil, diOH11,1218:19 in castor oil). Trihydroxy fatty acids were not detected in lesquerella oil. The differences in the structures of these C18 hydroxy fatty acids between lesquerella and castor oils indicated that the polyhydroxy fatty acids were biosynthesized and were not the result of autoxidation products.

Adipocere formation—The result of insufficient microbial degradation

Decomposition of human tissue is a microbial process. Microorganisms need certain environmental conditions to be able to grow and to degrade organic substances of tissue. Conditions that disturb degradation result in adipocere. Adipocere is a predominantly whitish, crumbly, inodorous, fatty feeling lipid bulk, which is originated mainly from human or animal fat. It may persist for many decades. Three reasons are repeatedly given for adipocere: formation of hydroxy fatty acids, a high amount of water in the environment of the corpse and lack of oxygen. Fatty acids can be exclusively degraded by respiration but not by fermentation. Microorganisms produce polyhydroxy fatty acids as storage substances comparable to human fat, in the presence of a surplus of organic compounds but a lack of other essential substances for the microbial activity like oxygen. In the end, formation of adipocere is the result of oxygen deprivation. In contact with oxygen adipocere can be degraded by microorganisms, but this is a long lasting process as oxygen exchange is very limited in soil and water.

Production of a Novel 9,12-Dihydroxy-10(E)-eicosenoic Acid from Eicosenoic Acid by Pseudomonas aeruginosa PR3

Microbial conversions of unsaturated fatty acids often generate polyhydroxy fatty acids, giving them new properties such as higher viscosity and reactivity. A bacterial strain Pseudomonas aeruginosa (PR3) has been intensively studied to produce mono-, di-, and trihydroxy fatty acids from different 9-cis-monoenoic fatty acids such as oleic acid, ricinoleic acid, and palmitoleic acid. However, from the results and the postulated similar metabolic pathways involved in these transformations, it was assumed that the enzyme system involved in transformation of the monoenoic fatty acid by strain PR3 could utilize fatty acids with different chain lengths and locations of the double bond. In this study was used as a substrate for bioconversion by strain PR3 eicosenoic acid (C20:1, ω-9) containing a singular cis double bond at different positions from the carboxyl end as oleic acid, and it was confirmed that PR3 could produce a novel 9,12-dihydroxy-10(E)-eicosenoic acid (DED) with 6.2% yield from eicosenoic acid. The structure of DED was confirmed using GC-MS, FTIR, and NMR analyses. DED production was maximized at 72 h after the substrate was added to the 24 h culture. Some other nutritional factors were also studied for optimal production of DED.

Synthesis and Characterization of Corn Oil Polyhydroxy Fatty Acids Designed as Additive Agent for Many Applications

AbstractBefore the advent of the modern food industry, vegetable oils (triglycerides) from many sources had a long history of use as condiments in cooking, personal care, and other therapeutic applications. Industrial applications of vegetable oils outside of food usage, on the other hand, have been limited on account of the shorter shelf‐life durability of these oils resulting from the natural unsaturation (carbon–carbon double bonds) in the structure of most triglycerides. In seeking to explore expanded utilization of this renewable resource, we have eliminated the above weakness by chemically modifying the double bonds in the material in an attempt to stabilize the oil. We have used FT‐IR and NMR spectroscopy to characterize the derivative whereas the physical and chemical properties of the product in terms of stability and flow characteristics have been investigated using differential scanning calorimetry (DSC), pressure DSC, rheometry and thermogravimetric analysis. In this modification of corn oil the data obtained indicate that the resulting poly‐hydroxylated acids are more stable than the native corn oil. Additionally, the obtained properties are unique and such that this product will be amenable to use in cosmetics, pharmaceuticals, and other industrial uses especially as a lubricity enhancing additive in fuel applications.

Identification of diacylglycerol and triacylglycerol containing 11,12,13-trihydroxy-9,14-octadecadienoic acid in castor oil

Castor oil has many industrial uses. Molecular species of acylglycerols containing monohydroxy, dihydroxy and trihydroxy fatty acids in castor oil have been reported. We report here the identification of acylglycerols containing a triOH18:2 fatty acid in castor oil. The structure of this novel fatty acid was proposed as 11,12,13-trihydroxy-9,14-octadecadienoic acid by the mass spectrometry of the lithiated adducts of acylglycerols in the HPLC fractions of castor oil. The fragmentation pathways of the lithiated adduct of 11,12,13-trihydroxy-9,14-octadecadienoic acid were proposed. We also proposed the biosynthetic pathways of polyhydroxy fatty acids in castor.

Synthesis and Characterization of the Intrinsic Properties of Milkweed Polyhydroxy Fatty Acids

AbstractSeed oils consist mainly of triglycerides, that is, they comprise a unit of glycerol backbone esterified with three acyl groups (usually but not limited to C16–C18) which may be saturated or unsaturated with one or more olefinic functionalities per acyl group. Very rarely do seed oils contain additional functional groups, such as hydroxyls as in castor and lesquerella seeds. Milkweed seed oil follows the natural triglyceride patterns, but with a difference in being highly poly olefinic. This character allows for the introduction of different reactive groupings into the structure of the oil so as to be amenable to tailoring to a variety of uses. Synthesis of the milkweed polyhydroxy triglyceride (MWPHTG) from the polyoxirane triglyceride derivative of milkweed oil using in situ peroxy acid epoxidation of the oil was previously reported. Subsequent acidolysis of the epoxy derivative gave the MWPHTG. Here the polyhydroxy triglyceride was saponified for glycerol removal thus generating the polyhydroxy fatty acids of milkweed oil. Studies of the physical characteristics, flow and stability of the resulting hydroxylated fatty acids using FTIR, NMR, DSC, Rheometry and TGA indicate a stable material with unique properties that would be useful as additives in many applications such as pharmaceuticals.

Self‐assembled polyhydroxy fatty acids vesicles: a mechanism for plant cutin synthesis

Despite its biological importance, the mechanism of formation of cutin, the polymeric matrix of plant cuticles, has not yet been fully clarified. Here, for the first time, we show the participation in the process of lipid vesicles formed by the self-assembly of endogenous polyhydroxy fatty acids. The accumulation and fusion of these vesicles (cutinsomes) at the outer part of epidermal cell wall is proposed as the mechanism for early cuticle formation.

Polyhydroxy Fatty Acids Derived from Sophorolipids

AbstractStarting from 17‐hydroxyoleic acid, which is readily available from acid alcoholysis of sophorolipids, several new polyhydroxy fatty acids have been synthesized. These compounds contain from 2 to 4 hydroxy groups, in some instances combined with other functional groups. The added hydroxy groups can be incorporated in the C18 chain in a variety of geometries, for example spaced widely throughout the chain at C1, C8, and C17. This regiochemical control will be of use in structure/function studies involving materials constructed from these hydroxy fatty acids. A further benefit is that the hydroxy groups can be present in protected or free states. The principal reactions used to introduce extra hydroxy groups are selenium oxide‐mediated allylic hydroxylation, osmium‐catalyzed dihydroxylation, and borohydride reduction of a carboxylic ester. These new compounds are expected to be of use in a number of areas, but particularly as building blocks for polymers or components of lubricant formulations.

Production of polyhydroxy fatty acids from linoleic acid by Clavibacter sp. ALA2

AbstractHydroxy fatty acids are important industrial materials. We isolated a microbial culture, Clavibacter sp. ALA2, which converts linoleic acid to many polyhydroxy fatty acids. Structures of the products were determined as: 12,13,17‐trihydroxy‐9(Z)‐octadecenoic (THOA, main product), 12‐[5‐ethyl‐2‐tetrahydrofuranyl]‐7,12‐dihydroxy‐9(Z)‐dodecenoic (ETDDA), and 12‐[5‐ethyl‐2‐tetrahydrofuranyl]‐12‐hydroxy‐9(Z)‐dodecenoic (ETHDA) acid. The yield of THOA was 25% and the relative amount of the products were THOA/ETDDA/ETHDA =9:1.3:1. The structures of the hydroxy unsaturated fatty acids resemble those of plant self‐defense substances.

Polyhydroxy fatty acids and their derivatives from plant oils

AbstractA novel process for the industrial production of hydroxylated fatty acids involves epoxidation of plant oils and their derivatives, followed by catalytic epoxy ring opening in the presence of water or other hydrogen donors, such as alcohols, diols, and amines. Depending on the starting material, epoxidation followed by opening of the oxirane ring leads to fatty acids that contain vicinal diol groups or to other substituted hydroxylated fatty acid derivatives. As an example for the preparation of a substituted hydroxylated fatty acid derivative, the reaction of epoxidized rapeseed oil with monobutylamine as hydrogen donor is described. Apart from the intended formation of hydroxyl groups with vicinal aminoalkyl groups, partial aminolysis of the ester compound was also observed. Another example describes the reaction of epoxidized rapeseed oil with different molar proportions of 1,4‐butanediol as hydrogen donor. Depending on the molar proportion of the hydrogen donor, interesterification, or intermolecular ether formation were observed as side reactions. The properties of various technical hydroxylated fatty acids and their derivatives, prepared according to this novel process, are given, and potential applications of these products are suggested.

The formation of oxo‐ and hydroxy‐fatty acids in heated fats and oils

AbstractA fast‐food fat (mostly tallow), olive oil and safflower oil were heated in air for 4 d and periodically analyzed for oxofatty acids (OFA), monohydroxy‐fatty acids (HFA) and polyhydroxy‐fatty acids (PHFA). After transmethylation, the OFA were estimated as 2,4‐dinitrophenylhydrazones, and the HFA and PHFA were quantitated as pyruvic acid 2,6‐dinitrophenylhydrazone esters. At least half of the maximum concentration attained for OFA, HFA and PHFA was generated between 16–24 h of heating of each oil. Safflower oil contained greater concentrations of HFA and PHFA than either olive oil or the fast‐food fat. The fastfood fat sample contained a greater concentration of OFA than did the other oils. The sum of the concentrations of OFA, HFA and PHFA at the time of maximum formation in the oils was approximately 260 μmoles/g at 48–72 h for safflower, 200 μmoles/g at 48–72 h for olive and 170 μmoles/g at 72 h for the fast‐food fat.