Fatty acids of <Emphasis Type="Italic">Stachys milanii</Emphasis> seeds

For the first time the total acid composition of the seeds of Stachys milanii Petrovic, a species endemic to the central and eastern Balkan Peninsula and growing spontaneously at two localities, was examined by GC and GC/MS. The major fatty acid was linoleic or oleic acid. Besides fatty acids that represent the usual seed oil constituents, 6-octadecynoic and 9-oxononanoic acid were identified in considerable amounts. The high content of 6-octadecynoic acid makes S. milanii seed oil a good potential source for the active substance in antifungal preparations. From the chemotaxonomical point of view, small amounts of octadecatrienoic acid detected in the seeds suggested classification of S. milanii as an archaic taxon. Published in Khimiya Prirodnykh Soedinenii, No. 4, pp. 315–317, July–August, 2007.     

Fatty acid,tocopherol, and sterol content of three <Emphasis Type="Italic">Teucrium</Emphasis> species from Tunisia

In the present study analyses concerning the composition of vitamin E, sterols, triglycerides, and fatty acids of three Teucrium species (Teucrium alopecurus, T. nabli, and T. polium) seed oil were performed. Linoleic, linolenic, and palmitic were the major fatty acids. The oil was characterized by a high amount of phytosterol, wherein clerosterol, sitosterol, and stigmasterol are the main constituents. The amount of tocopherol is nearly 550 mg/kg of oil, with α-tocopherol as the major isomer. Information concerning the composition of Teucrium seed oil is very important for evaluating the therapeutic effect of this oil. Published in Khimiya Prirodnykh Soedinenii, No. 3, pp. 261–264, May–June, 2009.     

Trichobitacin — a new ribosome-inactivating protein I. The isolation,physicochemical and biological properties of trichobitacin

From the press-residue of the fresh root tuber of Trichosanthes kirilowii Maim (Cucurbitaceae), a new ribosome-inactivating protein (RIP), trichobitacin, was isolated. It has the activity of RNA N-glycosidase and can inhibit the growth of human placental trophoblastic cells. Its molecular weight is 27,228 Da (ES-MS) and pI 9.6. It is a single chain basic RIP. Its amino acid composition was determined. It is a new RIP. It consists of 0.7～0.9% galactose and may be a glycoprotein. Its N- and C-terminal amino acid is Asp and Ala, respectively. Its N-terminal preliminary amino acid sequence has been determined.     

Characterization of <Emphasis Type="Italic">Opuntia ficus indica</Emphasis> seed oil from Tunisia

The lipid fraction of Opuntia ficus indica seeds was extracted and analyzed for its chemical and physical properties such as acid value, free fatty acid percentage (% FFA), iodine index, peroxide value, and saponification value as well as refractive index and density. The yield of seed oil was calculated as 11.75%. The acid and free fatty acid values indicated that the oil has a fairly low acidity. The triacylglycerols, fatty acids, sterols, and tocopherols were identified and their concentrations determined. The main TAGs were LLL (25.60%), OLL (21.53%), PLL (15.53%), and POL + SLL (12.73%). Linoleic acid (60.69%) was the dominant fatty acid, followed by oleic (21.42%) and palmitic (12.76%) acids, respectively. A high level of sterols making up 16.06 g/kg seed oil was present. The sterol marker, β-sitosterol, accounted for 71.60% of the total sterol content in the seed oil. Among the tocopherols present in the oil, γ-tocopherol (421.08 mg/kg) was the main constituent.     

Composition of essential oils from <Emphasis Type="Italic">Salvia anatolica</Emphasis>, a new species endemic from Turkey

The component composition of essential oils produced by steam distillation from flower heads, leaves, and stems of Salvia anatolica (Lamiaceae), a recently described new species endemic from Turkey, was studied by GC/FID and GC/MS. A total of 127 volatile components representing 96% of the oil was identified in essential oil from flower heads and leaves. It was found that the principal oil components of flower heads and leaves were α-pinene (10.9%), β-pinene (6.7%), α-copaene (6.3%), heptacosane (6.2%), and hexadecanoic acid (5.0%). A total of 109 volatile compounds representing 87.9% of the oil was characterized in essential oil isolated from stems. The principal oil components of stems were identified as hexadecanoic acid (27.2%), tetradecanoic acid (15.2%), dodecanoic acid (5.5%), and α-copaene (5.0%). __________ Translated from Khimiya Prirodnykh Soedinenii, No. 6, pp. 552–555, November–December, 2007.     

The chemical composition of fruits of <Emphasis Type="Italic">Pistacia atlantica</Emphasis> desf. subsp. <Emphasis Type="Italic">atlantica</Emphasis> from Algeria

The chemical composition of the fruits of the north algerian ecotype Pistacia atlantica subsp. atlantica was determined and compared to other fruits of different species in the genus growing in south Algeria and other Mediterranean regions. These fruits were analyzed for their dry matter, protein, crude oil, ash, fatty acids, and phytosterol content. The main fatty acids identified by gas chromatography were oleic (54.15%), linoleic (28.84%), and palmitic (12.21%) acids. The fruits of the north ecotype were found to be rich in protein, oil, fiber, and unsaturated fatty acids, suggesting that they may be valuable for food uses. The sterols isolated were campesterol, stigmasterol, β-sitosterol, and Δ⁵-avenasterol with β-sitosterol as the major constituent (85%±0.85). The biochemical data indicated an elevated MUFA rate (∼56%) in pistacia oil which may be important against certain pathologies for its nutritional and preventive virtues. Published in Khimiya Prirodnykh Soedinenii, No. 2, pp. 103–105, March–April, 2007.     

Change on storage of biological activity of <Emphasis Type="Italic">Viburnum opulus</Emphasis> seed components

Data for changes on storage under natural conditions for 11 months of moisture, oil content, acid and hydroperoxide number, and protein content of ground seed of Viburnum opulus were reported. The amino-acid and fatty-acid compositions were determined.Translated from Khimiya Prirodnykh Soedinenii, No. 5, pp. 349–351, September–October, 2004.     

<Superscript>1</Superscript>H NMR spectra of water contained in solutions of poly(<Emphasis Type="Italic">N</Emphasis>-vinylpyrrolidone) and its modification products in CDCl<Subscript>3</Subscript>

¹H NMR spectra in CDCl₃ of poly(N-vinylpyrrolidone), epoxidized poly(N-vinyl-pyrrolidone), and products derived from the latter by modification with amino acids (glycine, β-alanine, γ-aminobutyric acid, and ε-aminocaproic acid) were examined. The ¹H NMR spectra of the modified polymers contain signals for water protons due to different centers of water sorption. These signals differ in chemical shift and integral intensity and indicate a changed spatial packing of the polymer as the result of its modification. Published in Russian in Izvestiya Akademii Nauk. Seriya Khimicheskaya, No. 10, pp. 2377–2380, October, 2005.     

Flow-injection solid surface lanthanide-sensitized luminescence sensor for determination of <Emphasis Type="Italic">p</Emphasis>-aminobenzoic acid

A single optosensing device based on lanthanide-sensitized luminescence was developed for determination of p-aminobenzoic acid (PABA). The method is based on the formation of a complex between PABA and Tb(III) immobilized on the solid phase (QAE A-25 resin) placed inside the flow cell. NaCl (1 M) was used as carrier solution and HCl (0.05 M) as eluent. The sample solutions of PABA (100 μL) containing Tb(III) and buffered at pH = 6.0 were injected into the carrier stream and the luminescence was measured at λ _ex = 290 nm and λ _em = 546 nm. The method shows a linear range from 0.2 to 6.0 μg mL⁻¹ with an RSD of 1.2% (n = 10) and a sampling frequency of 22 h⁻¹. A remarkable characteristic of the method is its high selectivity which allows it to be satisfactorily applied to the analysis of PABA in pharmaceutical samples without prior treatment. Figure Typical emission bands of Tb(III) in a solid-phase PABA–Tb(III) luminescence spectrum     

Study on Inclusion Complex Formation of <Emphasis Type="Italic">m</Emphasis>-Aminobenzoic Acid with Native and Substituted <Emphasis Type="Italic">β</Emphasis>-Cyclodextrins

The complex formation of native and substituted β-cyclodextrins with m-aminobenzoic acid in water was characterized by calorimetry, ¹H NMR and UV spectroscopic studies. These studies showed that β-, hydroxypropyl-β- and methyl-β-cyclodextrins form 1:1 inclusion complexes with m-aminobenzoic acid. The thermodynamic properties of complex formation (K,Δ_c G ^o,Δ_c H ^o,Δ_c S ^o) were calculated. It was found that the processes of complexation are mainly favorable entropically. Introduction of hydroxypropyl- and methyl-substituents into the β-CD molecule results in negligible enhancement of stability of the complexes formed. The structure of these substituents has no influence on the stability constant values. The insertion of the carboxylic group of m-aminobenzoic acid into the cyclodextrin cavity was confirmed by ¹H NMR data.     

Biosynthetic production of universally <Superscript>13</Superscript>C-labelled polyunsaturated fatty acids as reference materials for natural health product research

Long-chain polyunsaturated fatty acids (LCPUFA) including eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) have become important natural health products with numerous proven benefits related to brain function and cardiovascular health. Not only are omega-3 fatty acids available in a plethora of dietary supplements, but they are also increasingly being incorporated as triglycerides into conventional foods, including bread, milk, yoghurt and confectionaries. Recently, transgenic oil seed crops and livestock have been developed that enhance omega-3 fatty acid content. This diverse array of matrices presents a difficult analytical challenge and is compounded further by samples generated through clinical research. Stable isotope ¹³C-labelled LCPUFA standards offer many advantages as research tools because they may be distinguished from their naturally abundant counterparts by mass spectrometry and directly incorporated as internal standards into analytical procedures. Further, ¹³C-labelled LCPUFAs are safe to use as metabolic tracers to study uptake and metabolism in humans. Currently, ¹³C-labelled LCPUFAs are expensive, available in limited supply and not in triglyceride form. To resolve these issues, marine heterotrophic microorganisms are being isolated and screened for LCPUFA production with a view to the efficient biosynthetic production of U-¹³C-labelled fatty acids using U-¹³C glucose as a carbon source. Of 37 isolates obtained, most were thraustochytrids, and either DHA or omega-6 docosapentaenoic acid (22:5n-6) were produced as the major LCPUFA. The marine protist Hyalochlorella marina was identified as a novel source of EPA and omega-3 docosapentaenoic acid (22:5n-3). As proof of principle, gram-level production of ¹³C-labelled DHA has been achieved with high chemical purity ( >99%) and high ¹³C incorporation levels (>90%), as confirmed by NMR and MS analyses. Finally, U-¹³C-DHA was enzymatically re-esterified to glycerol to yield a ¹³C-labelled tridocosahexaenoin.     

Chemical Constituents of Brown Rice Grain (<Emphasis Type="Italic">Oryza sativa</Emphasis>)

One new compound 3,7,11,15,19-pentamethyl-9α,10α,11α,17α,18α-pentahydroxy-n-tetracosan-1-oxy-p-hydroxycaffeoate (oryzaterpenyl caffeoate) (1), together with three known fatty acids linoleic acid, stearic acid and myristic acid were isolated and identified from the rice grain of Oryza sativa. The structure of the new compound was elucidated by 1D and 2D NMR spectroscopic techniques (¹H-¹HCOSY, ¹H-¹³C HETCOR) aided by EI-MS, and IR spectra. __________ Published in Khimiya Prirodnykh Soedinenii, No. 6, pp. 535–537, November–December, 2005.     

The influence of vegetable oils on biosurfactant production by Serratia marcescens

The production of biosurfactant, a surface-active compound, by two Serratia marcescens strains was tested on minimal culture medium supplemented with vegetable oils, considering that it is well known that these compounds stimulate biosurfactant production. The vegetable oils tested included soybean, olive, castor, sunflower, and coconut fat. The results showed a decrease in surface tension of the culture medium without oil from 64.54 to 29.57, with a critical micelle dilution (CMD⁻¹) and CMD⁻² of 41.77 and 68.92 mN/m, respectively. Sunflower oil gave the best results (29.75 mN/m) with a CMD⁻¹ and CMD⁻² of 36.69 and 51.41 mN/m, respectively. Sunflower oil contains about 60% of linoleic acid. The addition of linoleic acid decreased the surface tension from 53.70 to 28.39, with a CMD⁻¹ of 29.72 and CMD⁻² of 37.97, suggesting that this fatty acid stimulates the biosurfactant production by the LB006 strain. In addition, the crude precipitate surfactant reduced the surface tension of water from 72.00 to 28.70 mN/m. These results suggest that the sunflower oil’s linoleic acid was responsible for the increase in biosurfactant production by the LB006 strain.     

Citric acid production from raw glycerol by acetate mutants of Yarrowia lipolytica

Three acetate mutants of the yeast species Yarrowia lipolytica were screened using batch cultivation. The strain Y. lipolytica 1.31 was found to be the most suitable for citric acid production from raw glycerol, a by-product of biodiesel production from rapeseed oil. At the initial concentration of glycerol of 200 g dm⁻³, the citric acid production of 124.5 g dm⁻³, yield of 0.62 g g⁻¹, and productivity of 0.88 g dm⁻³ h⁻¹ were achieved. Presented at the 33rd International Conference of the Slovak Society of Chemical Engineering, Tatranské Matliare, 22–26 May 2006.     

<Emphasis Type="Italic">Rhizopus stolonifer</Emphasis> LAU 07: a novel source of fructosyltransferase

Increasing awareness of the importance of fructooligosaccharides (FOS) as ingredients of functional foods has led to intensive search of new sources of fructosyltransferases (FTase), enzymes responsible for the conversion of sucrose to fructooligosaccharides. A local strain of Rhizopus stolonifer isolated from spoilt orange fruit with high fructosyltransferase activity (U _t) of 12.31–45.70 U mL⁻¹ during a fermentation period of 24–120 h is herein reported. It showed low hydrolytic activity (U _h) in the range of 0.86–1.78 U mL⁻¹ during the same period. FOS yield of 34 % (1-kestose, GF₂, nystose, GF₃) was produced by FTase obtained from a 72 h-old culture using 60 g of sucrose per 100 mL of the substrate. When the isolate was grown in a defined submerged medium, its pH dropped sharply from the intial value of 5.5 to 1.0 within 24 h, and this value was maintained throughout the fermentation. The biomass content ranged from 8.8 g L⁻¹ at 24 h of fermentation to reach the maximum of 10 g L⁻¹ at 72 h. It was reduced to 5.6 g L⁻¹ at the end of 120 h of fermentation. This report represents the first reference to a strain of Rhizopus as a source of FTase for the production of FOS. The high U _t/U _h ratio shown by this isolate indicates that it may be a good strain for the industrial and commercial production of FOS. However, there is a need of further optimization of the bioprocess to increase the conversion efficiency of sucrose to FOS by the enzyme.     

Chemical characterization of unconventional palm oils from Hyophorbe indica and two other endemic Arecaceae species from Reunion Island

Abstract

Chemical characteristics of novel seed oils, yet not investigated, from three endemic Arecaceae (palm) species from Reunion Island are described. Fatty acid profiles are performed using two-dimensional gas chromatography-mass spectrometry. Carotenoid contents are determined by high performance liquid chromatography-mass spectrometry. The results of the investigations emphasize the particular composition of the unconventional red seed oil from Hyophorbe indica. Characteristic features of this oil reveal a high degree of unsaturation (50% of polyunsaturated fatty acids, with a high content (17%) of omega-3), which is possibly a unique fatty acid composition in the Arecaceae family. The two other palm oils from Dictyosperma album and Latania lontaroides contain high level of saturated fatty acids very similar to that of the edible palm oil. H. indica oil is also very rich in valuable carotenoids; in particular, lutein, β-carotene and lycopene are detected in a high content (respectively 45, 23 and 35?mg.kg^?1 in oil).     

Lipids from <Emphasis Type="Italic">Crambe amabilis</Emphasis> and <Emphasis Type="Italic">C. kotschyana</Emphasis> seeds grown with saline irrigation water

Lipids from Crambe amabilis Butk. et Majlun and C. kotschyana Boiss. grown in the open in Uzbekistan in soil imported from the Aral Sea region were studied. It was found that the seed oil content decreased with increasing salinity of irrigation water with EC 1.5 up to 9.0 dS/m. The content of polar lipids increased. The content of unsaturated acids from C. amabilis neutral lipids was 94.74–97.46%; C. kotschyana, 95.76– 96.78%.     

New glycosidic and other constituents from hulls of <Emphasis Type="Italic">Oryza sativa</Emphasis>

Three new compounds, 4-hydroxymethylene-7-(9,9,13-trimethylcyclohexyl)-heptanyl-3′,7′,7′-trimethylcyclohexa-2′,4′-dien-1′-oate (1), 1-(n-hexadec-7-enoxy)-6-(n-octadecanoxy)-β-D-glucopyranoside (2), and (Z)-12-hydroxy-9-octadecenoic acid-12-β-D-glucopyranoside (3), along with the known compound hexacosanoic acid (4), were isolated and identified from the rice hulls of Oryza sativa. Their structures were elucidated by 1D and 2D NMR spectroscopic techniques (¹H-¹H COSY, ¹H-¹³C HETCOR, DEPT) aided by EIMS, FABMS, HRFABMS, and IR spectra. Published in Khimiya Prirodnykh Soedinenii, No. 4, pp. 344–347, July–August, 2007.     

Intensification of Biocatalytical Processes by Synergistic Substrate Conversion. Fungal Peroxidase Catalyzed <Emphasis Type="Italic">N</Emphasis>-Hydroxy Derivative Oxidation in Presence of 10-Propyl Sulfonic Acid Phenoxazine

Many industrial pollutants, xenobiotics, and industry-important compounds are known to be oxidized by peroxidases. It has been shown that highly efficient peroxidase substrates are able to enhance the oxidation of low reactive substrate by acting as mediators. To explore this effect, the oxidation of two N-hydroxy derivatives, i.e., N-hydroxy-N-phenyl-acetamide (HPA) and N-hydroxy-N-phenyl-carbamic acid methyl ester (HPCM) catalyzed by recombinant Coprinus cinereus (rCiP) peroxidase has been studied in presence of efficient substrate 3-(4a,10a-dihydro- phenoxazin-10-yl)-propane-1-sulfonic acid (PPSA) at pH 8.5. The bimolecular constant of PPSA cation radical reaction with HPA was estimated to be (2.5 ± 0.2)·10⁷ M⁻¹ s⁻¹ and for HPCM was even higher. The kinetic measurements show that rCiP-catalyzed oxidation of HPA and HPCM can increase up to 33,000 times and 5,500 times in the presence of equivalent concentration of high reactive substrate PPSA. The mathematical model of synergistic rCiP-catalyzed HPA–PPSA and HPCM–PPSA oxidation was proposed. Experimentally obtained rate constants were in good agreement with those calculated from the model confirming the synergistic scheme of the substrate oxidation. In order to explain the different reactivity of substrates, the docking of substrates in the active site of the enzyme was calculated. Molecular dynamic calculations show that the enzyme–substrate complexes are structurally stable. The high reactive PPSA exhibited higher affinity to enzyme active site than HPA and HPCM. Furthermore, the orientation of HPA and HPCM was not favorable for proton transfer to the distal histidine, and different substrate reactivity was explained by these diversities.     

A kinetic method for <Emphasis Type="Italic">para</Emphasis>-nitrophenol determination based on its inhibitory effect on the catalatic reaction of catalase

The inhibitory effect of para-nitrophenol on the catalytic reaction of catalase was investigated. Michaelis-Menten kinetic parameters were determined from Lineweaver-Burk plots obtained in the absence or in the presence of the inhibitor. The inhibitor pattern, revealed by the Lineweaver-Burk plots, suggested a fully mixed inhibition mechanism. Spectrophotometric monitoring of the indicator reaction: in conjunction with initial rate measurements was employed for the kinetic determination of the inhibitor. Calibration plots of initial rate vs. para-nitrophenol concentration were linear in the concentration range 0.9·10⁻⁵–2.5·10⁻⁵ mol/L and the detection limit was 3·10⁻⁶ mol/L (417 μg/L) para-nitrophenol. Interferences from other phenolic compounds like orto-cresole, meta-and orto-nitrophenol were observed.