Infraspecific chemical variability of the leaf essential oil of Juniperus phoenicea var. turbinata from Portugal

The essential oil composition of 68 individual plants of Juniperus phoenicea from Portugal was investigated by GC, GC-MS and ¹³C NMR. -Pinene, β-phellandrene, -terpinyl acetate and myrcene were found to be the main constituents. Botanical and chemical data as well as phytogeographical distribution indicate J. phoenicea var. turbinata as the unique subspecies occurring in Portugal. Nevertheless, this taxon exhibits chemical polymorphism. The results of the oil compositions were processed by hierarchical clustering and principal component analysis allowing to establish three groups of essential oils differentiated by the content of -pinene, β-phellandrene and -terpinyl acetate.     

Use of gas chromatograms of essential leaf oils to compare eight taxa of genus Angophora (Myrtaceae): possible relationships to the genus Eucalyptus

Essential oils were extracted from leaves of eight taxa of the genus Angophora, and then analysed. As expected the individual components of these oils were essentially the same as those found in the Eucalyptus species of our earlier studies (Bignell et al., 1997b. Flavour Fragrance J. 12, 423-432). In addition, as is also the case with the bloodwood eucalypts, only relatively low yields of oil were obtained. In all cases the Cineole component was extremely small, but the oils of six of the eight contained very large concentrations of the sesquiterpene Bicyclogermacrene. A table of the 52 major oil components is included. Principal components analysis (PCA) was performed on the gas chromatograms (GC) of the essential oils and the resulting scores plots compared with the cladistic classification of Thiele and Ladiges (1988). Cladistics 4, 23–42). Because of the close relationship between genus Angophora and Eucalyptus ‘subgenus’ Corymbia, the GC data for the eight Angophora taxa were combined with corresponding data for eleven randomly chosen taxa from ‘subgenus’ Corymbia (Bignell et al., 1996b, Flavour Fragrance J. 11, 339–347; 1997a, Flavour Fragnance J. 12, 277–284) and a PCA performed on the total system.     

Essential oil composition of two endemic Centaurea species from Turkey, Centaurea mucronifera and Centaurea chrysantha, collected in the same habitat

The essential oils of two endemic Centaurea species from Turkey, C. mucronifera and C. chrysantha, collected in the same habitat, have been studied. The main compounds of the former were germacrene D (29.3%), β-eudesmol (17.4%) and β-caryophyllene (7.3%), while in the latter germacrene D (27.4%), caryophyllene oxide (9.5%) and bicyclogermacrene (5.4%) were detected among its major constituents. The two species produced many similar compounds in their essential oils that could be justified by the similar ecological conditions of their habitat, but also many differences were found that could confirm their taxonomic separation.     

Diversity in cyclic sesquiterpene production by Gossypium hirsutum

Major sesquiterpene components of oil of Texas Race Stock 810 of Gossypium hirsutum were - and β-selinene. This is the seventh cyclic terpene type found to date in this genus. Both - and β-selinene, along with aromadendrene, were found but only as minor components of extracts of several domestic cultivars of G. hirsutum.     

Composition and infraspecific variability of essential oil from Thymus herba barona Lois

The composition of the essential oils of individual plants of Thymus herba barona Lois. growing wild in Corsica was investigated by GC, GC/MS an carbon-13 NMR. Eight groups of essential oil were distinguished: (i) thymol, (ii) carvacrol, (iii) linalool, (iv) geraniol, (v) -terpenyl acetate, (vi) terpinen-4-ol, (vii) carvone and cis-dihydrocarvone. Three chemotypes -thymol, carvacrol and linalool – are common in the genus Thymus, two others – geraniol, -terpenyl acetate – are scarce, while the latest three ones – terpinen-4-ol, carvone and cis-dihydrocarvone are quite original. It is the first time that the cis-dihydrocarvone chemotype is described in the genus Thymus     

Stearidonic acid (18:4ω3) in Primula florindae

Stearidonic acid (18:4ω3), which is reported to be of rare occurrence in the plant kingdom and which is of considerable dietary and pharmaceutical interest has been found in three closely related Primula species. It occurs, together with γ-linolenic acid (3–4% of the seed oil total fatty acids), in significant percentages in Primula florindae (11%), P. sikkimensis (14%) and P. alpicola (14%). 18:4(ω3 may also be of chemotaxonomic interest in the genus Primula, as high levels may be typical for section Sikkimensis. The only commercial plant source of stearidonic acid known so far is the seed oil of Ribes nigrum.     

Antimicrobial and insecticidal activities of essential oil isolated from Turkish Salvia hydrangea DC. ex Benth.

The hydrodistillated essential oil of Salvia hydrangea was analyzed by GC–MS. Fifty-four different components representing 95.9% of the compounds in the oil were identified. Camphor (54.2%), -humulene (4.0%), cis-sesquisabinene hydrate (2.8%), myrtenol (2.6%), β-bisabolol (2.2%) and 1,8-cineole (2.1%) were found to be predominant components. The oil was also characterized by relatively high amount of oxygenated monoterpenes (69.6%). The oil was tested for fungitoxic effects against 33 agricultural pathogenic fungi using in vitro microbial growth inhibition assays. The oil exhibited considerable antifungal activity against a broad spectrum of tested fungi. Antibacterial activity of the oil was determined against 30 bacterial strains using the disc diffusion method. The oil had a very wide spectrum of antibacterial activity. However, it was not as active as penicillin. The oil showed 68.3–75.0% mortality against adults of Sitophilus granarius and Tribolium confusum, the major pests of wheat and wheat products, respectively. It can be concluded that the oil of S. hydrangea has a potential against agricultural pathogenic fungi and two stored pests, S. granarius and T. confusum.     

Inhibitory effects of different quantitative compositions of Hymenaea leaf resins on a generalist herbivore Spodoptera exigua

The hypotheses that quantitative variation in leaf resins in the leguminous genus Hymenaea may partially be a response to insect predation was tested in feeding experiments with the generalist herbivore, the beet armyworm, Spodoptera exigua Hübn (Noctuidae). Leaf resins of all Hymenaea species are comprised of essentially the same suite of sesquiterpene hydrocarbons, but discrete quantitative patterns have been classified as Compositional Types based on the amounts of major components. In this work pure extracts of leaf resins of Type I (high - and β-selinene), Type II (intermediate amounts of -and β-selinene and caryophyllene) and Type III (high caryophyllene) were incorporated into an artificial diet for the insects at 1 and 3.2% (dry wt). Resin Type treatments produced differential dose-dependent effects on growth rate (lower larval weights and increased time to pupation) and in mortality. More significant inhibitory effects occurred in Compositional Types with a predominance of a single compared (i.e. Types I and III). Significantly higher mortality occurring in Type III treatments suggests that caryophllene may have higher potential toxicity than - and β-selinene for S. exigua. These experiments indicate that feeding by generalist herbivores could be a factor determining quantitative compositional variation: (a) among populations; (b) during the development of leaves; (c) in the spatial distribution within the leaves; and (d) between parent tree and seedling progeny.     

Chemical composition of essential oils of seven species of Eugenia from Monteverde, Costa Rica

The leaf essential oils of seven species of Eugenia from Monteverde, Costa Rica (Eugenia austin-smithii, Eugenia cartagensis, Eugenia haberi, Eugenia monteverdensis, Eugenia zuchowskiae, Eugenia sp. A aff. haberi, and Eugenia sp. B aff. oerstediana) have been obtained by hydrodistillation and analyzed by GC–MS. The seven species were compared to determine the similarities and differences among their volatile chemical compositions. The major component in each of the seven species was as follows: E. austin-smithii and E. cartagensis was trans-2-hexenal, E. haberi and E. zuchowskiae was -pinene, E. monteverdensis was linalool, Eugenia sp. A was zingiberene, and Eugenia sp. B was 1,8-cineole. The following six components were present in all seven species: -copaene, β-caryophyllene, -humulene, δ-cadinene, trans-nerolidol, and torreyol. The complex array and differing abundances of these compounds among the Eugenia species studied suggest that they may provide useful characters in understanding the phylogenetic relationships among closely related species.     

Antiviral activity of the volatile oils of Melissa officinalis L. against Herpes simplex virus type-2

Melissa officinalis L. (Lamiaceae) has been used in a variety of practical applications in medical science. Our objective in the current study was to determine the effects of the volatile oil components of M. officinalis on Herpes simplex virus type 2 (HSV-2) replication in HEp-2 cells. Four different concentrations (25, 50, 100, 150 and 200 μg/ml) of volatile oils were examined. Experiments were carried out using HEp-2 cells. M. officinalis volatile oil was found to be non-toxic to HEp-2 cells up to a concentration of 100 μg/ml. It was, however, found to be slightly toxic at a concentration over of 100 μg/ml. The antiviral activity of non-toxic concentrations against HSV-2 was tested. The replication of HSV-2 was inhibited, indicating that the M. officinalis L. extract contains an anti-HSV-2 substance.     

Phytosterols from Murraya exotica

From the cyclohexane extract of the leaves of Murraya exotica, five novel phytosterols: (23S)-23-ethyl-24-methyl-cycloart-24(24¹)-en-3β-ol; 3β-methoxy-(23S)-23-ethyl-24-methyl-cycloart-24(24¹)-en-3β-ol; (23 S)-23-ethyl-24-methyl-cycloart-24(24¹)-3β-yl acetate; (23ξ)-23-isopropyl-24-methyl-cycloart-25-en-3β-ol and (23ξ)-23-isopropyl-24-methyl-cycloart-25-en-3β-yl acetate have been isolated. Structural elucidation of the isolated compounds is based on physical, chemical and spectral analysis including IR, ¹H and ¹³CNMR and mass spectrometry.     

Fcγ receptor IIA and IIIB polymorphisms are associated with susceptibility to cerebral malaria

Human FcγRIIA and FcγRIIIB exhibit genetic polymorphisms, FcγRIIA-131H/R and FcγRIIIB-NA1/NA2, coding for different capacities for IgG binding and phagocytosis. Recently, FcγRIIA-131R was reported to be associated with protection against high-density Plasmodium falciparum infection in Kenya. Furthermore, FcγRIIIB-NA1/NA2 polymorphism was shown to influence FcγRIIA function in an allele-specific manner. In this study, we examined a possible association of FcγRIIA-131H/R and FcγRIIIB-NA1/NA2 polymorphisms with malaria severity in 107 cerebral malaria patients, 157 non-cerebral severe malaria patients, and 202 mild malaria controls living in northwest Thailand. This study reveals that, with the FcγRIIIB-NA2 allele, the FcγRIIA-131H/H genotype is associated with susceptibility to cerebral malaria (OR 1.85, 95% CI 1.14–3.01; P=0.012), although these polymorphisms are not individually involved in the disease severity. Our results suggest that FcγRIIA-131H/R and FcγRIIIB-NA1/NA2 polymorphisms have an interactive effect on host defense against malaria infection.     

Oxidation of 1,4-alkanediols into γ-lactones via γ-lactols using Rhodococcus erythropolis as biocatalyst

Whole cells of Rhodococcus erythropolis DSM 44534 grown on ethanol, (R)- and (S)-1,2-propanediol were used for biotransformation of racemic 1,4-alkanediols into γ-lactones. The cells oxidized 1,4-decanediol (1a) and 1,4-nonanediol (2a) into the corresponding γ-lactones 5-hexyl-dihydro-2(3H)-furanone (γ-decalactone, 1c) and 5-pentyl-dihydro-2(3H)-furanone (γ-nonalactone, 2c), respectively, with an EE(R) of 40–75%. The transient formation of the γ-lactols 5-hexyl-tetrahydro-2-furanol (γ-decalactol, 1b) and 5-pentyl-tetrahydro-2-furanol (γ-nonalactol, 2b) as intermediates was observed by GC–MS. 1,4-Pentanediol (3a) was transformed into 5-methyl-dihydro-2(3H)-furanone (γ-valerolactone, 3c) whereas (R)- and (S)-2-methyl-1,4-butanediol (4a) was converted to the methyl-substituted γ-butyrolactones 4-methyl-dihydro-2(3H)-furanone (4c₁) and 3-methyl-dihydro-2(3H)-furanone (4c₂) in a ratio of 80:20 with a yield of 55%. Also cis-2-buten-1,4-diol (5a) was transformed resulting in the formation of 2(5H)-furanone (γ-crotonolactone, 5c). At the higher pH values of 8.8 the yield of lactone formed was improved; however, the enatiomeric excesses were slightly higher at the lower pH of 5.2.     

Cellulase production during growth of Talaromyces emersonii CBS 814.70 on lactose containing media

The thermophilic fungus Talaromyces emersonii CBS 814.70 is capable of growth on lactose containing media. The cell protein produced towards the end of growth on that substrate is similar to those levels produced during growth of the organism on cellulose. During growth of the organism on lactose, cellulase is secreted into the medium. Analysis of the components of the cellulase system shows that both β-glucosidase and endoglucanase enzymes are produced. Levels of β-glucosidase produced during growth of the organism on lactose are well in excess of levels of that enzyme produced at any time during growth of the organism on cellulose, and we have shown that the form of that enzyme produced during growth on lactose is β-glucosidase III (BG-III). Analysis of the forms of endoglucanase indicates that not all forms of enzyme produced during growth on cellulose are produced during growth on lactose. β-Galactosidase activity was found to be present in the mycelial associated fraction, though our evidence suggests that this may simply be an incidental activity of the cell associated form of β-glucosidase IV (BG-IV).     

Botryosphaeran, a new substrate for the production of β-1,3-glucanases by Botryosphaeria rhodina and Trichoderma harzianum Rifai

Botryosphaeria rhodina and Trichoderma harzianum Rifai were grown on botryosphaeran (an exopolysaccharide (EPS) of the β-1,3;1,6-d-glucan type produced by B. rhodina) as sole carbon source with the objective of producing β-glucanases of the 1,3-type. Conditions for β-1,3-glucanase production by T. harzianum were examined by a statistical response surface method, and showed maximal enzyme production at 5 days growth in media containing 1.5 g/l of EPS. Good agreement was obtained between the experimental values of β-1,3-glucanase activity and the corresponding values predicted by the mathematical model. The crude β-1,3-glucanase preparations were active towards a number of different β-1,3-glucans and β-glucosides. The mycelium of B. rhodina also proved to be a good substrate for β-1,3-glucanase production by both fungal species.     

Terpenes from the essential oil of Cymbopogon distans

A GC/MS study of the hydrocarbon fraction and the fraction containing oxygenated compounds showed the presence of 12 monoterpene hydrocarbons (28.4%), 13 sesquiterpene hydrocarbons (32.8%), 3 sesquiterpene alcohols (27.2%), 2 esters (7.2%) and 3 carbonyl compounds (4.4%) in the essential oil of Cymbopogon distans. Of these, 27 compounds have been identified.     

Furanoeremophilanes from Gynoxys species

Five new furanoeremophilanes were isolated from three Gynoxys species. The structures of three 3,6β-diacyloxyfuranoeremophilanes previously isolated from G. sancto-antonii have to be revised to the corresponding 1β,6β derivatives.     

Catalytic function and affinity purification of site-directed mutant β-cyclodextrin glucanotransferase from alkalophilic Bacillus firmus var. alkalophilus

Subsites −3 and −7 in the active site of β-cyclodextrin glucanotransferase (β-CGTase) from alkalophilic Bacillus firmus var. alkalophilus were modified through site-directed mutagenesis to obtain novel mutant CGTases. Four mutant CGTases, H59Q, Y96M, 90-PPI-92, and Δ(154–160) were constructed and produced using a recombinant E. coli with a secretive expression system extracellularly. The secreted mutant β-CGTases were purified by one-step affinity adsorption chromatography using a β-cyclodextrin (CD) polymer as an adsorbent to nearly homogeneous purity. The catalytic activities were modified significantly compared to the wild-type. In particular, the Y96M and Δ(154–160) mutants increased cyclization activity around 1.5 times without any significant reduction of coupling and hydrolyzing activities. Meanwhile, the Y96M and Δ(154–160) mutants exhibited a much higher conversion yield into CDs from 28.6 to 39% without any recognizable change in the CD ratio. The conversion yield into linear maltooligosaccharides was also significantly reduced. The catalytic functions of subsites −3 and −7 in the active site of β-CGTase would appear to be related to the overall productivity of CDs rather than the product specificity.     

Organization of 3β-hydroxysteroid dehydrogenase/isomerase and cytochrome P450scc into a catalytically active molecular complex in bovine adrenocortical mitochondria

We have previously reported the co-localization [Cherradi et al., Endocrinology 134 (1994) 1358–1364] of 3β-hydroxysteroid dehydrogenase/isomerase (3β-HSD) and cytochrome P450_scc (cyt. P450_scc) in the inner membrane and in the intermembrane contact sites of adrenocortical mitochondria. This observation raises the question of a possible functional association between the two proteins. Isolated bovine adrenocortical mitochondria are able to convert cholesterol to progesterone without the need of exogenous cofactors. An association of 3β-HSD and cyt. P450_scc is observed during the purification of 3β-HSD from mitochondria. The behaviour of 3β-HSD on a column of Heparin-Sepharose is modified by the presence of cyt. P450_scc. Immunoprecipitations from mitochondria with either anti-cyt. P450_scc or anti 3β-HSD antibodies result in a co-precipitation of the two proteins. Both proteins engaged in these immunocomplexes are catalytically active. The interaction was further demonstrated by the surface plasmon resonance method using purified components. An affinity constant of 0.12 μM between 3β-HSD and P450_scc was obtained. These observations suggest that P450_scc and 3β-HSD may associate into a molecular complex in the mitochondrial compartment and may constitute a functional steroidogenic unit, thus opening new possibilities in the regulation of the production of progesterone and its flow in the adrenocortical cell.