Microbial transformation of 12-hydroxyoctadecanoic acid to 5-n-hexyl-tetrahydrofuran-2-acetic acid

Stationary-phase cells of a corynebacterium (FUI-2) and a bacillus (NRRL B-14864) isolate, when grown aerobically in 1% YE medium at 25°C, converted 12-hydroxystearic acid to a major compound, 5-n-hexyl-tetrahydrofuran-2-acetic acid, and other intermediate and minor compounds (6-hydroxydodecanoic acid, 4-hydroxydecanoic acid, 4-ketodecanoic acid and γ-decanolactone). The yields of 5-n-hexyl-tetrahydrofuran-2-acetic acid, 4-hydroxydecanoic acid, and γ-decanolactone, byBacillus lentus NRRL B-14864 were 43%, 18% and 5%, respectively, after 2.5 d of incubation. Presented in part at the 3rd Annual Student Research Conference, Board of Governors Universities, University Park, Illinois, April 1992; in part at the 92nd American Society for Microbiology General Meeting, New Orleans, Louisiana, May 1992; and in part at the 35th West Central States Biochemistry Conference Annual Meeting, Manhattan, Kansas, October 1992.     

Biotransformation of 12-hydroxyoctadecanoic acid to 12-hydroxyoctadecanamide by Bacillus cereus 50

A new microbial isolate (Bacillus cereus 50) transformed 12-hydroxyoctadecanoic acid to 12-hydroxyoctadecanamide when grown aerobically in 1% yeast extract medium at 30°C, shaken at 250 rpm for 2 to 5 d. The compound was purified by thin-layer chromatography and characterized by infrared, gas chromatography, mass spectrometry, and nuclear magnetic resonance. The yields of 12-hydroxyoctadecanamide were 9.1 and 21.5% after 2 and 5 d, respectively.     

A pronounced catalytic activity of heteropoly compounds supported on dealuminated USY for liquid-phase esterification of acetic acid with n-butanol

12-Phosphotungstic acid and its cesium salts supported on a dealuminated ultra-stable Y zeolite were prepared, and showed the high catalytic activity in the liquid-phase esterification of acetic acid with n-butanol. The supported Cs_2.5H_0.5PW₁₂O₄₀ catalyst gave a high conversion of n-butanol of 94.6% and a selectivity for n-butyl acetate of 100%, accompanying the high water-tolerance and catalytic reusability without regeneration.     

Synthesis and surface properties of chemodegradable anionic surfactants: Sodium carboxylates of cis-1,3-dioxane derivatives

The 2-n-alkyl-5-carboxy-5-methyl-1,3-dioxanes were obtained in good yield from the reaction of aliphatic aldehydes with 2,2-bis(hydroxymethyl)propionic acid in dichloromethane solution, catalyzed by p-toluenesulfonic acid monohydrate. Nuclear magnetic resonance analysis indicated that they were pure cis-isomers with the axial configuration of the carboxylic group at the C-5 carbon atom of the 1,3-dioxane ring. The acids were converted, with retention of the configuration, to their sodium salts by reaction with sodium methoxide or sodium hydroxide in methanol. The physicochemical properties of the acids and sodium salts, as well as their surface properties at the aqueous solution-air interface, were determined. Critical micelle concentration, surface excess concentration, surface area demand per molecule of sodium salts at the monomolecular surface layer, and standard free energy of micellization were determined based on surface tension measurements. Part XXXIV in the series: Chemical Structure and Surface Activity. Part XXXIII: A. Piasecki, A. Sokolowski, B. Burczyk, R. Gancarz, and U. Kotlewska, Synthesis, Surface Properties and Hydrolysis of Chemodegradable Anionic Surfactants: Diastereomerically Pure cis- and trans-2,5-Disubstituted-1,3-dioxanes. J. Colloid Interface Sci. 192:74–82 (1997).     

Synthesis and surface properties of chemodegradable anionic surfactants: Diastereomeric (2-n-alkyl-1,3-dioxan-5-yl) sulfates with monovalent counter-ions

Sodium, potassium and ammonium cis- and trans-(2-n-alkyl-1,3-dioxan-5-yl) sulfates 6–8 (alkyl: n-C₉H₁₉, 6a–8a, and n-C₁₁H₂₃, 6b–8b) were synthesized in a reaction of aliphatic aldehydes 1a,b with glycerol 2 followed by separation in high yields of individual geometric isomers of cis-and trans-2-n-alkyl-5-hydroxy-1,3-dioxanes, cis-3a,b and trans-3a,b, followed by sulfation with sulfur trioxide-pyridine complex, and finally neutralization with NaOH, KOH, and NH₄OH, respectively. Physical data of the compounds and some surface properties of 2-n-nonyl derivatives, such as critical micelle concentration (CMC), effectiveness of aqueous surface tension reduction (Π_CMC), surface excess concentration Γ_CMC, and the surface area demand per molecule (A_CMC), were determined. It was shown that the surface activity of these compounds is influenced both by their geometric structure and by the monovalent counter-ion.     

A novel technique for the preparation of secondary fatty amides II: The preparation of ricinoleamide from castor oil

The butyl amide of ricinoleic acid (N-n-butyl-12-hydroxy-(9Z)-octadecenamide) was prepared from a neat mixture of castor oil andn-butylamine (fatty ester/amine molar ratio, 1:1.3). No catalyst was required. The identity and purity of the amide was assessed by thin-layer chromatography and confirmed by elemental analysis and by infrared and C¹³ nuclear magnetic resonance spectroscopy. High product yields were achieved at 45 and 65°C in 48 and 20 h, respectively. The reaction was inhibited by the addition of trimethylpentane and dioxane, but not by water. An attempt was made to prepare the amide from methyl ricinoleate, rather than castor oil; even after 10 d only partial conversion was achieved. Attempts to prepare the amide from methyl-n-butylamine, rather thann-butylamine, were also unsuccessful. The ease with which secondary fatty amides can be produced from an oil that consists primarily of the glycerol esters of hydroxylated fatty acids indicates that the described procedure has industrial utility.     

Survey of the fatty acid composition of peanut (arachis hypogaea) germplasm and characterization of their epoxy and eicosenoic acids

Peanut (Arachis hypogaea) plant introductions (732) were analyzed for fatty acid composition. Palmitate varied from 8.2 to 15.1%, stearate 1.1 to 7.2%, oleate 31.5 to 60.2%, linoleate 19.9 to 45.4%, arachidate 0.8 to 3.2%, eicosenoate 0.6 to 2.6%, behenate 1.8 to 5.4%, and lignocerate 0.5 to 2.5%. The eicosenoate was shown to be cis-11-eicosenoate. In addition, epoxy fatty acids were found in many plant introductions in percentages ranging as high as 2.5%. These were tentatively identified as chiefly 9,10-epoxy stearate and coronarate with smaller amounts of vernolate. The percentage of palmitate was shown to be correlated positively with linoleate and negatively with oleate, eicosenoate, and lignocerate. Stearate was highly correlated with arachidate and negatively with eicosenoate and lignocerate. Oleate and linoleate, the two major fatty acids, were negatively correlated. Arachidate was negatively correlated with eicosenoate, and eicosenoate was positively correlated with lignocerate. Behenate and lignocerate were positively correlated. Epoxy esters were positively correlated with palmitate and negatively with oleate. Segregation of the plant introductions by axis flower, growth habit, and pod types showed significant differences that reflected the same fatty acid groupings revealed by the correlations.     

Production of 15-, 16- and 17-hydroxy-9-octadecenoic acids by bioconversion of oleic acid withBacillus pumilus

SeveralBacillus strains were tested for their ability to hydroxylate oleic acid. Two strains—BD-174 and BD-226—converted oleic acid into a trio of hydroxy-octadecenoic acids. Bioconversion in 72–120 hr produced 5–11% of hydroxy acids relative to oleic acid as measured by gas chromatography. These acids were identified as the 15-, 16- and 17-hydroxy-9-octadecenoic acids by gas chromatography-mass spectrometry of trimethyl silyl derivatives of the product acids and their hydrogenated counterparts.     

Synthesis and surface properties of chemodegradable anionic surfactants: Sodium (2-n-alkyl-1,3-dioxan-5-yl)sulfates

In the reaction of cis- and trans-2-n-alky-5-hydroxyl-1,3-dioxane mixtures with SO₃ · pyridine complex, followed by neutralization with sodium hydroxide or sodium carbonate, a new group of anionic surfactants, i.e., sodium cis- and trans-(2-n-alkyl-1,3-dioxan-5-yl)sulfates were obtained. The hydrophobic intermediates used in the sulfation reaction were obtained in high yields from four-component glycerol acetals by the process of transacetalization and selective crystallization of 1,3-dioxane derivatives. The physical data of the new compounds and some of their surface properties, such as critical micelle concentration (CMC), effectiveness of water surface tension reduction π_CMC, standard free energies of adsorption and micellization, ΔG _ads ^o and ΔG _CMC ^o , surface excess concentration, Γ_CMC, and the surface area demand per molecule, A_CMC, were determined. It was shown that the surface activity of the standard anionic surfactant sodium dodecyl sulfate should be similar to the surface activity of sodium (2-n-decyl-1,3-dioxan-5-yl)sulfate. Part XXX in the series: Chemical Structure and Surface Activity. Part XXIX: Sokołowski, A., B. Burczyk, and H.-R. Holzbauer, Adsorption of n-butyloligooxypropylene-pentaoxyethylene block copolymers at the aqueous solution-air interface, Collid Surfaces, in press.     

Biocatalytic production of 10-hydroxystearic acid, 10-ketostearic acid, and their primary fatty amides

The objective of this study was to develop scaleup bioprocesses for producing 10-hydroxystearic acid (10-HSA) and 10-ketostearic acid (10-KSA) as well as their primary amides for potential new uses. A reactor process was examined to obtain the mono-oxygenated FA using Sphingobacterium thalpophilum (NRRL B-14797) and Bacillus sphaericus (NRRL NRS-732), which solely produce 10-HSA and 10-KSA, respectively, from technical-grade oleic acid. By using an 8-h-old B-14797 culture grown in a manganese-containing WF6 medium, pH 7.3, at 28°C under 350 rpm agitation and 0–50% dissolved oxygen concentrations provided by a controlled sparger aeration, the production of 10-HSA reached 7 g/L with a 40% yield in 4 d. In using a 12-h-old NRS-732 culture grown in a pyruvate-containing PF6 medium, pH 6.5, at 30°C under 750 rpm agitation without any sparger aeration during the conversion reaction, 10-KSA production reached 7.9 g/L with a yield of more than 54% in 72 h. The scaleup reactor process provided crystalline 10-HSA and 10-KSA for producting new primary amides via a lipase-catalyzed amidation reaction with yields of 94 and 92%, respectively. The primary amides of 10-HSA and 10-KSA displayed m.p of 115 and 120°C, respectively.     

N-acyl-L-leucines of biologically active uncommon fatty acids: Synthesis and antibacterial activity

The effect of structural variation in the fatty acid chain on the antibacterial activity ofN-acyl-L-leucines was investigated. PureN-acyl leucines of some structurally different and biologically active uncommon fatty acids were synthesized for the first time and tested for antimicrobial activity.N-Stearoyl-,N-oleoyl- andN-ricinoleoyl leucines were also evaluated for comparison. TheN-acyl leucines exhibited greater activity in acid form than the methyl ester form and against gram positive bacteria than gram-negative bacteria. The presence of a cyclopropane or a hydroxy group or unsaturation in the acyl chain increased the antibacterial activity. Shifting the hydroxyl group toward the amide linkage resulted in a diminished effect on the antibacterial activity.     

Monooxygenase system of Bacillus megaterium ALA2: Studies on linoleic acid epoxidation products

Bacillus megaterium ALA2 produces many oxygenated FA from linoleic acid: 12,13-dihydroxy-9(Z)-octadecenoic acid; 12,13,17-trihydroxy-9(Z)-octadecenoic acid; 12,13,16-trihydroxy-9(Z)-octadecenoic acid; 12-hydroxy-13,16-epoxy-9 (Z)-octadecenoic acid; and 12,17;13,17-diepoxy-16-hydroxy-9 (Z)-octadecenoic acid. Recently, we studied the monooxygenase system of B. megaterium ALA2 by comparing its palmitic acid oxidation products with those of the well-studied catalytically self-sufficient P450 monooxygenase of B. megaterium ATCC 14581 (NRRL B-3712) and of B. subtilis strain 168 (NRRI B-4219). We found that their oxidation products are identical, indicating that their monooxygenase systems (hydroxylation) are similar. Now, we report that strain ALA2 epoxidizes linoleic acid to 12,13-epoxy-9(Z)-octadecenoic acid and 9,10-epoxy-12 (Z)-octadecenoic acid, the initial products in the linoleic acid oxidation. The epoxidation enzyme did not oxidize specific double bond of the linoleic acid. The epoxidation activity of strain ALA2 was compared with the above-mentioned two Bacillus strains. These two Bacillus strain also produced 12,13-expoxy-9 (Z)-octadecenoic acid and 9,10-epoxy-12(Z)-octadecenoic acid, indicating that their epoxidation enzyme systems might be similar. The ratios of epoxy FA production by these three strains (A1 A2, NRRI B-3712, and NRRI B-4219) were, respectively, 5.56∶0.66∶0.18 for 12,13-epoxy-9(Z)-octadecenoic acid and 2.43∶0.41∶0.57 for 9,10-epoxy-12(Z)-octadecenoic acid per 50 mL medium per 48 h.     

Solvothermal Synthesis and Crystal Structure of a Novel 3D Triorganotin Polymer Containing Polyfunctional Ligand 5-mercapto-1(H)-tetrazoleacetic acid

A novel 3D tri-n-butyltin complex [(n-Bu₃Sn)₂(C₂H₂N₄SCO₂)]_n (1) has been solvothermally synthesized and structurally characterized by elemental analysis, FT-IR, NMR (¹H, ¹³C, and ¹¹⁹Sn) spectra and X-ray crystallography. X-ray data of complex 1 reveals that it is a 3D tri-n-butyltin coordination polymer with a metal-organic framework (MOF) structure , significant N → Sn interactions plays an important role in the construction of this structure.     

Cryptolepis buchnani seed oil: A rich source of keto fatty acid

A keto fatty acid (9-oxo-cis-12-octadecenoic acid) has been isolated in appreciable amounts (45.9%) fromCryptolepis buchnani seed oil. The identification was based on chemical and spectroscopic methods.     

Double-bond location in monounsaturated fatty acids by dimethyl disulfide derivatization and mass spectrometry: Application to analysis of fatty acids in pheromone glands of four lepidoptera

A rapid analytical procedure for the determination of the position of double bonds in mixtures of monounsaturated fatty acid methyl esters has been developed. The method is based on direct capillary GC-MS-EI analysis of dimethyl disulfide adducts. The procedure was applied to mixtures of monounsaturated fatty esters from pheromone gland extracts of three tortricids from theChoristoneura genus,C. fumiferana, C. occidentalis, andC. pinus pinus, and one noctuid from the Plusiinae subfamily,Plusia chalcites. A correlation was found between the known major pheromone components in the four species with the corresponding fatty acids. Some of the additional fatty acids may be precursors to as yet unidentified minor pheromone components, present in extremely small quantities, in these species.On sabbatical leave from the Institute of Plant Protection, Volcani Center, Bet Dagan 50 250, Israel.     

Biosynthesis of tetrahydrofuranyl fatty acids from linoleic acid by <Emphasis Type="Italic">clavibacter</Emphasis> sp. ALA2

Clavibacter sp. ALA2 converts linoleic acid into many novel oxygenated products including hydroxy FA and tetrahydrofuranyl unsaturated FA (THFA). One of them was tentatively identified by GC-MS as 12,13,16-trihydroxy-9(Z)-octadecenoic acid (12,13,16-THOA) (Hou, C.T., H.W. Gardner, and W. Brown, J Am. Oil Chem. Soc. 78∶1167–1169, 2001). We have separated and purified 12,13,16-THOA from its isomer, 12,13,17-THOA, by silica gel column chromatography and by preparative TLC. Its structure was then confirmed by proton and ¹³C NMR analyses. Purified 12,13,16-THOA was used as a substrate to study the biosynthesis of THFA. Within 24 h of incubation, cells of strain ALA2 converted 12,13,16-THOA to both 12-hydroxy-13,16-epoxy-9(Z)-octadecenoic acid (12-hydroxy-THFA) and 7,12-dihydroxy-13,16-epoxy-9(Z)-octadecenoic acid (7,12-dihydroxy-THFA). The relative abundance of 7,12-dihydroxy-THFA increased with incubation time, whereas that of 12,13,16-THOA and of 12-hydroxy-THFA decreased. Therefore, the biosynthetic pathway of THFA from linoleic acid by strain ALA2 is as follows: linoleic acid→12,13-dihydroxy-9(Z)-octadecenoic acid→12,13,16-THOA→12-hydroxy-THEA→7,12-dihydroxy-THFA.     

Kinetics of the formation of symmetrical wax esters from the corresponding alcohols with the use of hydrobromic acid and hydrogen peroxide

The primary aliphatic alcohols n-octanol, n-decanol, and n-dodecanol have been converted to their corresponding symmetrical esters by using HBr and H₂O₂ in the absence of a solvent. The reaction was carried out at 30, 40, and 50°C and at mole ratios of alcohol to HBr of 1∶0.1, 1∶0.2, 1∶0.3, and 1∶0.5. The rate of the reaction was found to increase with increase in the reaction temperature and concentration of HBr. The maximal conversion of n-octanol was 72% at 40°C and a mole ratio of n-octanol to HBr of 1∶0.5. The kinetics of the reaction have been established, and the reaction was found to be first-order with respect to alcohol and bromine concentration in the organic phase, and second-order with respect to both. The second-order rate constants for n-octanol, n-decanol, and n-dodecanol are 27.08, 32.58, and 37.42 mL mol⁻¹ min⁻¹, respectively, at 40°C. The activation energy for the esterification reaction of n-octanol was found to be 16.32 kcal mol⁻¹.     

Keggin型配合物[（CH2）5NH2]5BW12O40的合成及其催化氧化合成苯甲酸的研究

以钨酸钠、硼酸为原料合成母体酸,以六氢吡啶为有机配体合成了有机／无机杂化电荷转移配合物[（CH2）5NH2]5BW12O40。通过红外光谱、X-射线衍射和热重分析袁征,确认所合成的化合物中多酸阴离子仍保留Keggin结构。将新合成的[（CH2）5NH2]5BW12O40杂多酸哌啶盐应用到苯甲醛氧化合成苯甲酸反应中,考察了催化剂用量、氧化责4H2O2（质量分数为30％）的用量、反应温度、反应时间等对苯甲酸收率的影响。确定最佳工艺条件为：n（催化剂）：n（苯甲醛）=1．880×10-3：1;n（H2O2）：n（苯甲醛）=4．760：1;反应温度80℃;反应时间4h,此条件下苯甲酸的收率可达到80%。     

Enzymatic synthesis of high-purity structured lipids with caprylic acid at 1,3-positions and polyunsaturated fatty acid at 2-position

We attempted to synthesize high-purity structured triacylglycerols (TAG) with caprylic acid (CA) at the 1,3-positions and a polyunsaturated fatty acid (PUFA) at the 2-position by a two-step enzymatic method. The first step was synthesis of TAG of PUFA (TriP), and the second step was acidolysis of TriP with CA. Candida antarctica lipase was effective for the first reaction. When a reaction medium of PUFA/glycerol (3∶1, mol/mol) and 5% immobilized Candida lipase was mixed for 24 h at 40°C and 15 mm Hg, syntheses of TAG of γ-linolenic, arachidonic, eicosapentaenoic, and docosahexaenoic acids reached 89, 89, 88, and 83%, respectively. In these reactions, the lipase could be used for at least 10 cycles without significant loss of activity. In the second step, the resulting trieicosapentaenoin was acidolyzed at 30°C for 48h with 15 mol parts CA using 7% of immobilized Rhizopus delemar lipase. The CA content in the acylglycerol fraction reached 40 mol%. To increase the content further, the acylglycerols were extracted from the reaction mixture with n-hexane and were allowed to react again with CA under conditions similar to those of the first acidolysis. After three successive acidolysis reactions, the CA content reached 66 mol%. The content of dicapryloyl-eicosapentaenoyl-glycerol reached 86 wt% of acylglycerols, and the ratio of 1,3-dicapryloyl-2-eicosapentaenoyl-glycerol to 1(3),2-dicapryloyl-3(1)-eicosapentaenoyl-glycerol was 98∶2 (w/w). In this reaction, the lipase could be used for at least 20 cycles without significant loss of activity. Repeated acidolysis of the other TriP with CA under similar conditions synthesized 1,3-dicapryloyl-2-γ-linolenoyl-glycerol, 1,3-dicapryloyl-2-arachidonoyl-glycerol, and 1,3-dicapryloyl-2-docosahexaenoyl-glycerol in yields of 58, 87, and 19 wt%, respectively.     

Oxidation of toluene to benzoic acid over modified V2O5-TiO2 catalyst with SeO2, TeO2 and Sb2O3

It was found that the addition of SeO₂, TeO₂ or Sb₂O₃ to a V₂O₅/TiO₂ catalyst greatly improved the catalytic activity in the vapor phase oxidation of toluene to selectively form benzoic acid.