Pentadecanedioic acid production from 15-hydroxypentadecanoic acid using an engineered biocatalyst with a co-factor regeneration system

α,ω-Dicarboxylic acids are valuable precursors in various chemical industries and have recently been produced using biotechnological methods to overcome the limitations of chemical synthesis. Nonanedioic acid, decanedioic acid, undecanedioic acid, and dodecanedioic acid have been produced at high concentrations from ω-hydroxycarboxylic acids using engineered biocatalysts. However, no study has been attempted on the efficient production of pentadecanedioic acid. Here, the production of pentadecanedioic acid from 15-hydroxypentadecanoic acid was carried out with alcohol dehydrogenases (ADHs), aldehyde dehydrogenases (ALDHs), and NAD(P)H oxidases, including NAD(P)H flavin oxidoreductase (NFO), that used a co-factor regeneration system, derived from several species and expressed in Escherichia coli. Among the enzymes, Kangiella koreensis ADH (KkADH), Geobacillus kaustophilus ALDH (GkALDH), and Deinococcus radiodurans NFO (DrNFO) were selected because they had the highest activity. E. coli expressing pRSF-DrNFO and pACYC-KkADH/GkALDH as the best distribution of three genes in two plasmids was used as a biocatalyst to produce pentadecanedioic acid. The optimal conditions for producing pentadecanedioic acid from 15-hydroxypentadecanoic acid by the biocatalyst were pH 8.0, 35°C, 5% (v/v) methanol, 40 g L⁻¹ cells, and 60 mM 15-hydroxypentadecanoic acid with agitation at 250 rpm. Under these optimized conditions, 57.4 mM pentadecanedioic acid was produced after 3 h, with a molar yield of 95.6% and a productivity of 19.1 mM h⁻¹. The molar yield and concentration of pentadecanedioic acid showed the highest values among the reported biotechnological production of pentadecanedioic acid.     

Characterisation of CYP102A25 from Bacillus marmarensis and CYP102A26 from Pontibacillus halophilus: P450 Homologues of BM3 with Preference towards Hydroxylation of Medium‐Chain Fatty Acids

Cytochrome P450 monooxygenases are highly desired biocatalysts owing to their ability to catalyse a wide variety of chemically challenging C?H activation reactions. The CYP102A subfamily of enzymes are natural catalytically self‐sufficient proteins consisting of a haem and FMN‐FAD reductase domain fused in a single‐component system. They catalyse the oxygenation of saturated and unsaturated fatty acids to produce primarily ω?1, ω?2 and ω?3 hydroxy acids. These monooxygenases have potential applications in biotechnology; however, their substrate range is still limited and there is a continued need to add diversity to this class of biocatalysts. Herein, we present the characterisation of two new members of this class of enzymes, CYP102A25 (BMar) from Bacillus marmarensis and CYP102A26 (PHal) from Pontibacillus halophilus, both of which express readily in a recombinant bacterial host. BMar exhibits the highest activity toward myristic acid and shows moderate activity towards unsaturated fatty acids. PHal exhibits broader activity towards mid‐chain‐saturated (C₁₄–C₁₈) and unsaturated fatty acids. Furthermore, PHal shows good regioselectivity for the hydroxylation of myristic acid, targeting the ω?2 position for C?H activation.     

2′‐Deoxyribonucleoside Phosphoramidate Triphosphate Analogues as Alternative Substrates for E. coli Polymerase III

Thermostable bacterial polymerases like Taq, Therminator and Vent exo^? are able to perform DNA synthesis by using modified DNA precursors, a property that is exploited in several therapeutic and biotechnological applications. Viral polymerases are also known to accept modified substrates, and this has proven crucial in the development of antiviral therapies. However, non‐thermostable polymerases of bacterial origin, or engineered variants, that have similar substrate tolerance and could be used for synthetic biology purposes remain to be identified. We have identified the α subunit of Escherichia coli polymerase III (Pol III α) as a bacterial polymerase that is able to recognise and process as substrates several pyrophosphate‐modified dATP analogues in place of its natural substrate dATP for template‐directed DNA synthesis. A number of dATP analogues featuring a modified pyrophosphate group were able to serve as substrates during enzymatic DNA synthesis by Pol III α. Features such as the presence of potentially chelating chemical groups and the size and spatial flexibility of the chemical structure seem to be of major importance for the modified leaving group to play its role during the enzymatic reaction. In addition, we could establish that if the pyrophosphate group is altered, deoxynucleotide incorporation proceeds with an efficiency varying with the nature of the nucleobase. Our results represent a great step towards the achievement of a system of artificial DNA synthesis hosted by E. coli and involving the use of altered nucleotide precursors for nucleic acid synthesis.     

A Coenzyme‐Independent Decarboxylase/Oxygenase Cascade for the Efficient Synthesis of Vanillin

Vanillin is one of the most widely used flavor compounds in the world as well as a promising versatile building block. The biotechnological production of vanillin from plant‐derived ferulic acid has attracted much attention as a new alternative to chemical synthesis. One limitation of the known metabolic pathway to vanillin is its requirement for expensive coenzymes. Here, we developed a novel route to vanillin from ferulic acid that does not require any coenzymes. This artificial pathway consists of a coenzyme‐independent decarboxylase and a coenzyme‐independent oxygenase. When Escherichia coli cells harboring the decarboxylase/oxygenase cascade were incubated with ferulic acid, the cells efficiently synthesized vanillin (8.0 mM , 1.2 g L ^?1) via 4‐vinylguaiacol in one pot, without the generation of any detectable aromatic by‐products. The efficient method described here might be applicable to the synthesis of other high‐value chemicals from plant‐derived aromatics.     

Semirational Protein Engineering of CYP153AM.aq.‐CPRBM3 for Efficient Terminal Hydroxylation of Short‐ to Long‐Chain Fatty Acids

The regioselective terminal hydroxylation of alkanes and fatty acids is of great interest in a variety of industrial applications, such as in cosmetics, in fine chemicals, and in the fragrance industry. The chemically challenging activation and oxidation of non‐activated C?H bonds can be achieved with cytochrome P450 enzymes. CYP153A_M.aq.‐CPR_BM3 is an artificial fusion construct consisting of the heme domain from Marinobacter aquaeolei and the reductase domain of CYP102A1 from Bacillus megaterium. It has the ability to hydroxylate medium‐ and long‐chain fatty acids selectively at their terminal positions. However, the activity of this interesting P450 construct needs to be improved for applications in industrial processes. For this purpose, the design of mutant libraries including two consecutive steps of mutagenesis is demonstrated. Targeted positions and residues chosen for substitution were based on semi‐rational protein design after creation of a homology model of the heme domain of CYP153A_M.aq., sequence alignments, and docking studies. Site‐directed mutagenesis was the preferred method employed to address positions within the binding pocket, whereas diversity was created with the aid of a degenerate codon for amino acids located at the substrate entrance channel. Combining the successful variants led to the identification of a double variant—G307A/S233G—that showed alterations of one position within the binding pocket and one position located in the substrate access channel. This double variant showed twofold increased activity relative to the wild type for the terminal hydroxylation of medium‐chain‐length fatty acids. This variant furthermore showed improved activity towards short‐ and long‐chain fatty acids and enhanced stability in the presence of higher concentrations of fatty acids.     

Characterization of Carboxylic Acid Reductases for Biocatalytic Synthesis of Industrial Chemicals

Carboxylic acid reductases (CARs) catalyze the reduction of a broad range of carboxylic acids into aldehydes, which can serve as common biosynthetic precursors to many industrial chemicals. This work presents the systematic biochemical characterization of five carboxylic acid reductases from different microorganisms, including two known and three new ones, by using a panel of short‐chain dicarboxylic acids and hydroxy acids, which are common cellular metabolites. All enzymes displayed broad substrate specificities. Higher catalytic efficiencies were observed when the carbon chain length, either of the dicarboxylates or of the terminal hydroxy acids, was increased from C₂ to C₆. In addition, when substrates of the same carbon chain length are compared, carboxylic acid reductases favor hydroxy acids over dicarboxylates as their substrates. Whole‐cell bioconversions of eleven carboxylic acid substrates into the corresponding alcohols were investigated by coupling the CAR activity with that of an aldehyde reductase in Escherichia coli hosts. Alcohol products were obtained in yields ranging from 0.5 % to 71 %. The de novo stereospecific biosynthesis of propane‐1,2‐diol enantiomer was successfully demonstrated with use of CARs as the key pathway enzymes. E. coli strains accumulated 7.0 mm (R)‐1,2‐PDO (1.0 % yield) or 9.6 mm (S)‐1,2‐PDO (1.4 % yield) from glucose. This study consolidates carboxylic acid reductases as promising enzymes for sustainable synthesis of industrial chemicals.     

Fabrication of antibacterial silver nanoparticle‐modified chitosan fibers using Eucalyptus extract as a reducing agent

Green chemical method could be a promising route to achieve large scale synthesis of nanostructures for biomedical applications. Here, we describe a green chemical synthesis of silver nanoparticles (Ag NPs) on chitosan‐based electrospun nanofibers using Eucalyptus leaf extract. A series of silver salt (AgNO₃) amounts were added to a certain composition of chitosan/polyethylene oxide aqueous acetic acid solution. The solutions were then electrospun to obtain nanofibrous mats and then, morphology and size of nanofibers were analyzed by scanning electron microscopy (SEM). Incubation of AgNO₃‐containing mats into Eucalyptus leaf extract led to the formation of Ag NP clusters with average diameter of 91 ± 24 nm, depicted by SEM and transmission electron microscopy. Surface enhanced Raman spectroscopy also confirmed formation of Ag NPs on the nanofibers. The mats also showed antimicrobial activity against Escherichia coli and Staphylococcus aureus bacteria with bigger inhibition zone for extract‐exposed mats against S. aureus. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 42133.     

Chemical structure of poly(β‐cyclodextrin‐co‐citric acid)

We synthesized water‐insoluble polymers, poly(β‐cyclodextrin‐co‐citric acid)s, by heating a mixture of citric acid, cyclodextrin (CD), and Na₂HPO₄ as a catalyst with a 6 : 1 : 2 molar ratio at 160, 170, and 180°C for 10 and 20 min. The chemical composition of the polyesters was determined by high pressure liquid chromatography (HPLC) analysis of the polymer hydrolysates. The crosslinking mechanisms and thermal degradation of the polymers were also investigated. The polyesters contained 30–35% citric acid, 1–4% unsaturated carboxylic acids (i.e., itaconic, cis‐aconitic, trans‐aconitic, and mesaconic acids), and 60–70% CD, whereas about 40% of them were able to form inclusion complexes. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

Reactivity,chemical structure,and molecular mobility of urea–formaldehyde adhesives synthesized under different conditions using FTIR and solid‐state 13C CP/MAS NMR spectroscopy

This study was conducted to investigate the effects of reaction pH condition and hardener type on the reactivity, chemical structure, and molecular mobility of urea–formaldehyde (UF) resins. Three different reaction pH conditions, such as alkaline (7.5), weak acid (4.5), and strong acid (1.0), were used to synthesize UF resins, which were cured by adding four different hardeners (ammonium chloride, ammonium sulfate, ammonium citrate, and zinc nitrate) to measure gel time as the reactivity. FTIR and ¹³C‐NMR spectroscopies were used to study the chemical structure of the resin prepared under three different reaction pH conditions. The gel time of UF resins decreased with an increase in the amount of ammonium chloride, ammonium sulfate, and ammonium citrate added in the resins, whereas the gel time increased when zinc nitrate was added. Both FTIR and ¹³C‐NMR spectroscopies showed that the strong reaction pH condition produced uronic structures in UF resin, whereas both alkaline and weak‐acid conditions produced quite similar chemical species in the resins. The proton rotating‐frame spin–lattice relaxation time (T_1ρH) decreased with a decrease in the reaction pH of UF resin. This result indicates that the molecular mobility of UF resin increases with a decrease in the reaction pH used during its synthesis. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 88: 2677–2687, 2003     

Synthesis and characterization of bio‐based thermosetting resins from lactic acid and glycerol

A bio‐based thermoset resin has been synthesized from glycerol reacted with lactic acid oligomers of three different chain lengths (n): 3, 7, and 10. Lactic acid was first reacted with glycerol by direct condensation and the resulting branched molecule was then end‐functionalized with methacrylic anhydride. The resins were characterized by Fourier‐transform infrared spectroscopy (FT‐IR), by ¹³C‐NMR spectroscopy to confirm the chemical structure of the resin, and by differential scanning calorimetry and dynamic mechanical thermal analysis (DMTA) to obtain the thermal properties. The resin flow viscosities were also measured using a rheometer with different stress levels for each temperature used, as this is an important characteristic of resins that are intended to be used as a matrix in composite applications. The resin with a chain length of three had better mechanical, thermal, and rheological properties than the resins with chain lengths of seven and 10. Also, its bio‐based content of 78% and glass transition temperature of 97°C makes this resin comparable to commercial unsaturated polyester resins. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40488.     

Fully renewable limonene‐derived polycarbonate as a high‐performance alkyd resin

Limonene‐derived polycarbonate‐based alkyd resins (ARs) have been prepared by copolymerization of limonene dioxide with CO₂, catalysed by a β‐diiminate zinc–bis(trimethylsilyl)amido complex, and subsequent chemical modification with soybean oil fatty acids using triphenylethylphosphonium bromide as the catalyst. This quantitative partial modification was realized via epoxy–carboxylic acid chemistry, affording ARs with higher oil lengths, lower polydispersities and higher glass transition temperatures (T_g) in comparison to a conventional polyester AR based on phthalic acid, multifunctional polyol pentaerythritol and soybean fatty acid. The novel limonene polycarbonate AR and the conventional polyester AR were evaluated as coatings and both the physical drying (without the presence of the oxidative drying accelerator Borchi® Oxy Coat) and chemical curing (with Borchi® Oxy Coat) processes of these coatings were monitored by measuring the König hardness and complex modulus development with time. A better performance was obtained for the alkyd paint containing polycarbonates modified with fatty acids (FA‐PCs), which showed a faster chemical drying, a higher König hardness and a higher T_g in coating evaluation, demonstrating that the fully renewable FA‐PCs are promising resins for alkyd paint applications. © 2019 Society of Chemical Industry     

Influence of substrate dideuteration on the reaction of the bifunctional heme enzyme psi factor producing oxygenase A (PpoA)

PpoA is a bifunctional enzyme that catalyzes the dioxygenation of unsaturated C18 fatty acids. The products of this reaction are termed psi factors and have been shown to play a crucial role in conferring a balance between sexual and asexual spore development as well as production of secondary metabolites in the fungus Aspergillus nidulans. Studies on the reaction mechanism revealed that PpoA uses two different heme domains to catalyze two subsequent reactions. Initially, the fatty acid substrate is dioxygenated at C8, yielding an 8‐hydroperoxy fatty acid at the N‐terminal domain. This reaction is catalyzed by a peroxidase/dioxygenase‐type domain that exhibits many similarities to prostaglandin H2 synthases and involves a stereospecific homolytic hydrogen abstraction from C8 of the substrate. The C terminus harbors a heme thiolate P450 domain in which rearrangement of the 8‐hydroperoxide to the final product, a 5,8‐dihydroxy fatty acid, takes place. To obtain further information about the intrinsic kinetics and reaction mechanism of PpoA, we synthesized C5‐dideutero‐ and C8‐dideutero‐oleic acid by a novel protocol that offers a straightforward synthesis without employing the toxic additive hexamethylphosphoramide (HMPA) during C? C coupling reactions or mercury salts upon thioketal deprotection. These deuterated fatty acids were then employed for kinetic analysis under multiple‐turnover conditions. The results indicate that the hydrogen abstraction at C8 is the rate‐determining step of the overall reaction because we observed a KIE (V_H/V_D) of ～33 at substrate saturation that suggests extensive nuclear tunneling contributions for hydrogen transfer. Deuteration of the substrate at C5, however, had little effect on V_H/V_D but resulted in a different product pattern presumably due to an altered lifetime and partitioning of a reaction intermediate.     

Chemically and enzymatically catalyzed synthesis of C6–C10 alkyl benzoates

The esterification of benzoic acid with n‐hexanol, n‐octanol, 2‐ethyl hexanol and n‐decanol was investigated in detail. An analysis of the reaction kinetics of esterification in the presence of different commercially available chemical catalysts was carried out. The effects of catalyst type and loading on the reaction rate were studied. Although the considered reaction is bimolecular, it showed a first‐order behavior, and a linear dependence with respect to the catalyst concentration was observed. Hence, a new approach is presented to describe the reaction kinetics accurately over a wide range. The application of biotechnological synthesis applying different enzymes as catalysts offers an interesting alternative besides chemical synthesis. Especially an esterase from Bacillus subtilis immobilized on Sepabeads EC‐EP showed high stability and was applied for 2 days in the synthesis of hexyl benzoate. Nevertheless, the chemical reaction route remains superior with respect to the catalyst activities under the applied conditions, which were 25 kU/g for the chemical reaction and 0.7 kU/g for the best enzymatic conversion.     

Synthesis of polyaniline nanofiber and copolymerization with acrylate through in situ emulsion polymerization

Polyaniline (PANI) was prepared, respectively, by direct mixed oxidation method in different acids. Scanning electron microscopy showed that high quality of PANI nanofibers can be obtained easily in hydrochloric acid, sulfuric acid, and acetic acid, especially in the sulfuric acid; infrared and ultraviolet spectra characterization showed all products were the doped PANI. Then, using complex emulsifiers, PANI was dispersed in acrylate emulsion by supersonic dispersion assisted with mechanical stirred to obtain mixed pre‐emulsion, the result showed different PANI performed different dispersing stability in the pre‐emulsion. More importantly, PANI–polyacrylate copolymer was prepared through multi‐steps in situ emulsion polymerization using water‐soluble azo (VA‐044) as initiator. Experiment showed that good dispersing stability of PANI in the pre‐emulsion was premise to obtain the final stable copolymer emulsion. Further, the micro‐morphology and thermal property of the copolymer were studied by transmission electron microscopy, differential scanning calorimetry, and thermogravimetric analyzer. The result proved that acrylate occurred in situ polymerization on surface of PANI nanofibers, the presence of PANI increased glass transition temperature (T_g) and thermal decomposed temperature of the copolymer. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

β‐Aminopeptidase‐Catalyzed Biotransformations of β2‐Dipeptides: Kinetic Resolution and Enzymatic Coupling

We have previously shown that the β‐aminopeptidases BapA from Sphingosinicella xenopeptidilytica and DmpA from Ochrobactrum anthropi can catalyze reactions with non‐natural β³‐peptides and β³‐amino acid amides. Here we report that these exceptional enzymes are also able to utilize synthetic dipeptides with N‐terminal β²‐amino acid residues as substrates under aqueous conditions. The suitability of a β²‐peptide as a substrate for BapA or DmpA was strongly dependent on the size of the C_α substituent of the N‐terminal β²‐amino acid. BapA was shown to convert a diastereomeric mixture of the β²‐peptide H‐β²hPhe‐β²hAla‐OH, but did not act on diastereomerically pure β²,β³‐dipeptides containing an N‐terminal β²‐homoalanine. In contrast, DmpA was only active with the latter dipeptides as substrates. BapA‐catalyzed transformation of the diastereomeric mixture of H‐β²hPhe‐β²hAla‐OH proceeded along two highly S‐enantioselective reaction routes, one leading to substrate hydrolysis and the other to the synthesis of coupling products. The synthetic route predominated even at neutral pH. A rise in pH of three log units shifted the synthesis‐to‐hydrolysis ratio (v_S/v_H) further towards peptide formation. Because the equilibrium of the reaction lies on the side of hydrolysis, prolonged incubation resulted in the cleavage of all peptides that carried an N‐terminal β‐amino acid of S configuration. After completion of the enzymatic reaction, only the S enantiomer of β²‐homophenylalanine was detected (ee>99 % for H‐(S)‐β²‐hPhe‐OH, E>500); this confirmed the high enantioselectivity of the reaction. Our findings suggest interesting new applications of the enzymes BapA and DmpA for the production of enantiopure β²‐amino acids and the enantioselective coupling of N‐terminal β²‐amino acids to peptides.     

Studies on the effect of epoxide equivalent weight of epoxy resins on thermal,mechanical, and chemical characteristics of vinyl ester resins

Three samples of vinyl ester resins (VERs) were synthesized using bisphenol‐A‐based epoxy resins of varying epoxide equivalent weights (EEW) and acrylic acid in presence of triphenylphosphine as a catalyst at 80 ± 2°C. The cresyl glycidyl ether was used as reactive diluent during the synthesis of VERs. A suitable reaction mechanism was proposed and discussed for the reactions involving epoxide group and acid groups. This was further confirmed by infrared spectroscopic analysis. The maximum peak temperature from DSC were at 106.05°C, 114.20°C, and 128.86°C for benzoyl peroxide initiated VERs viz. samples V₁C_V, V₂C_V, and V₃C_V, respectively, increased with the increase of EEW of the parent epoxy resin. It has also been found that the films of VER having highest EEW of bisphenol‐A epoxy resin showed best chemical resistance amongst all other VERs in this study. The mechanical properties such as hardness and flexibility also showed a similar trend. The thermal stability was found to decrease with the increase of EEW of bisphenol‐A epoxy resin in the VERs. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010     

Detection of Unprecedented CYP74 Enzyme in Mammal: Hydroperoxide Lyase CYP74C44 of the Bat Sturnira hondurensis

The genome of the neotropical fruit bat Sturnira hondurensis was recently sequenced, revealing an unexpected gene encoding a plant-like protein, CYP74C44, which shares ca. 90% sequence identity with the putative CYP74C of Populus trichocarpa. The preparation and properties of the recombinant CYP74C44 are described in the present work. The CYP74C44 enzyme was found to be active against the 13- and 9-hydroperoxides of linoleic and α-linolenic acids (13-HPOD, 13-HPOT, 9-HPOD, and 9-HPOT, respectively), as well as the 15-hydroperoxide of eicosapentaenoic acid (15-HPEPE). All substrates studied were specifically transformed into chain cleavage products that are typical for hydroperoxide lyases (HPLs). The HPL chain cleavage reaction was validated by the identification of NaBH₄-reduced products (Me/TMS) of 15-HPEPE and 13- and 9-hydroperoxides as (all-Z)-14-hydroxy-5,8,11-tetradecatrienoic, (9Z)-12-hydroxy-9-dodecenoic, and 9-hydroxynonanoic acids (Me/TMS), respectively. Thus, CYP74C44 possessed the HPL activity that is typical for the CYP74C subfamily proteins.     

Structural Analysis of the Minor Cerebrosides from a Glass Sponge Aulosaccus sp.

The minor cerebrosides from a Far‐Eastern glass sponge Aulosaccus sp. were analyzed as constituents of some multi‐component RP‐HPLC fractions. The structures of eighteen new and one known cerebrosides were elucidated on the basis of NMR spectroscopy, mass spectrometry, optical rotation data and chemical transformations. These β‐D‐glucopyranosyl‐(1→1)‐ceramides contain sphingoid bases N‐acylated with straight‐chain (2R)‐2‐hydroxy fatty acids, namely, (2S,3S,4R,11Z)‐2‐aminoeicos‐11‐ene‐1,3,4‐triol, acylated with 15E‐22:1, 16Z‐21:1, 15Z‐21:1, 15Z‐20:1, 15E‐20:1, 19:0, 18:0 acids, (2S,3S,4R)‐2‐amino‐13‐methyltetradecane‐1,3,4‐triol—with 19Z‐26:1, 16Z‐23:1, 23:0, 22:0 acids, (2S,3S,4R)‐2‐amino‐14‐methylpentadecane‐1,3,4‐triol—with 16Z‐23:1, 16E‐23:1, 15Z‐22:1, 22:0 acids, (2S,3S,4R)‐2‐amino‐14‐methylhexadecane‐1,3,4‐triol, linked to 16Z‐23:1, 15Z‐22:1 acids, (2S,3S,4R)‐2‐amino‐9‐methylhexadecane‐1,3,4‐triol—to 16Z‐23:1 acid, and (2S,3S,4R)‐2‐aminohexadecane‐1,3,4‐triol, attached to 15Z‐22:1 acid. The 13‐methyl and 9‐methyl‐branched trihydroxy sphingoid base backbones (C₁₅ and C₁₇, respectively) have not been found previously in sphingolipids. The ceramide parts, containing other backbones, present new variants of N‐acylation of the marine sphingoid bases with the 2‐hydroxy fatty acids. The combination of the instrumental and chemical methods used in this study improved the efficiency of the structural analysis of such complex cerebroside mixtures that gave more detailed information on glycosphingolipid metabolism of the organism.     

Friedel–Crafts benzylation of aromatics with benzyl alcohols catalyzed by heteropoly acids supported on mesoporous silica

Heteropoly acids (HPA), such as tungstophosphoric acid (H₃PW₁₂O₄₀ · xH₂O) (HPW), molybdophosphoric acid (H₃PMo₁₂O₄₀ · xH₂O) (HPMo) and tungstosilicic acid (H₄SiW₁₂O₄₀ · xH₂O) (HSiW) were supported on mesoporous silica such as MCM‐41, FSM‐16 and SBA‐15 by the impregnation method to enhance the catalytic activity of these solid acids by their dispersion on the support with high surface area. These supported solid catalysts were used in the benzylation of benzene and substituted aromatics with benzyl alcohol (BnOH). The immobilization enhanced the catalytic performances and HPW supported on MCM‐41 exhibited the best activity for benzylation among the heteropoly acids, although dibenzyl ether (DBE) formation by the dehydration of BnOH also occurred. The mesoporous architecture of the silica enhances the activity of benzylation because of the high dispersion of HPW on the support with high surface area; however, no steric restriction by the pores of mesoporous silica was observed. The catalysts used in the present study retained their catalytic activity for five reaction cycles. The rate of benzylation of substituted benzenes and benzylating agents was influenced by the electronic nature of the substituent. Electron‐donating groups enhanced the rate of reaction; however, electron‐withdrawing groups retard the benzylation. These results show that the reactivity of benzene and benzyl alcohols was retarded by electron‐withdrawing groups. The formation of polybenzylated products was influenced by the reactivity of diphenylmethane products. The benzylation accompanies the DBE formation by the dehydration of BnOH, particularly in the initial stages, because the benzylation of aromatics with BnOH is not as rapid as the dehydration of BnOH. However, direct benzylation of benzene occurs with DBE and DBE participates in the benzylation of benzene via BnOH after its hydrolysis. This is further supported by the effect of water on the benzylation of benzene by DBE, although there is a possibility of direct benzylation of DBE with benzene. Copyright © 2006 Society of Chemical Industry     

Physicochemical and thermal characterization of seed oil from Mexican mamey sapote <Emphasis Type="Italic">(Pouteria sapota)</Emphasis>

The aim of this research was to perform a physicochemical characterization of native mamey sapote seed oil [Pouteria sapota (Jacq.) H.E. Moore and Stearn]. Mamey sapote seed oil showed good oxidative stability as it had low peroxide, free fatty acid, and p‐anisidine values. The main fatty acids present in the oil were palmitic, stearic and oleic acid, constituting five major triacylglycerides families: PLP, POP, StOO, POSt and StOSt. Crystallization and melting points of the oil were ?37.7 and 23.84 °C, respectively. The oil had higher SFC when the temperature was lower than 10 °C. X‐ray diffraction patterns showed that prolonged storage times lead to the formation of β crystals. Micrographs showed granular crystals (91–105 μm), with needle edges similar to cocoa butter. In addition, mamey sapote seed oil can be used in confectionery products or as a possible substitute for cocoa butter to improve and obtain good‐quality products.