Individual Compounds (individual + compound)

Distribution by Scientific Domains

Selected Abstracts

Berry anthocyanins: isolation, identification and antioxidant activities,

Marja P Kähkönen
Abstract Anthocyanins from bilberry, blackcurrant and cowberry were isolated for antioxidant evaluation. Individual compounds were identified and quantified using HPLC and HPLC/ESI,MS techniques. Antioxidant and radical-scavenging capacities of the isolates were studied in emulsified methyl linoleate and human low-density lipoprotein (LDL) in vitro and in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) test. The total anthocyanin contents in the phenolic extracts of bilberry, blackcurrant and cowberry were 6000, 2360 and 680 mg kg,1 fresh weight respectively. There were four dominant compounds in blackcurrant (glucosides and rutinosides of cyanidin and delphinidin), three in cowberry (monoglycosides of cyanidin) and 15 in bilberry (monoglycosides of cyanidin, delphinidin, malvidin, peonidin and petunidin). Quantification as cyanidin-3-glucoside equivalents gave markedly lower results regarding the total anthocyanin concentration and the content of individual delphinidin and malvidin compounds compared with quantification based on corresponding standard compounds. Berry anthocyanins were highly active radical scavengers in the DPPH test and effective antioxidants in emulsion and human LDL. Copyright © 2003 Society of Chemical Industry [source]

Phenolic compounds, lycopene and antioxidant activity in commercial varieties of tomato (Lycopersicum esculentum)

Isabel Martķnez-Valverde
Abstract Nine commercial varieties of tomato (Rambo, Senior, Ramillete, Liso, Pera, Canario, Durina, Daniella and Remate) produced in Spain were analysed for their lycopene content, content of phenolic compounds and antioxidant capacity. The phenolic compounds were characterised as flavonoids (quercetin, kaempferol and naringenin) and hydroxycinnamic acids (caffeic, chlorogenic, ferulic and p -coumaric acids). Antioxidant activity was measured using the DPPH and ABTS assays. The concentrations of lycopene and the various phenolic compounds as well as the antioxidant activity were significantly influenced by the tomato variety. Quercetin, the most abundant flavonoid, was found in concentrations ranging between 7.19 and 43.59,mg,kg,1 fresh weight, while naringenin levels were lower than 12.55,mg,kg,1. The most abundant hydroxycinnamic acid was chlorogenic acid, with values ranging from 14 to 32,mg,kg,1 fresh weight, followed by caffeic acid, while p -coumaric and ferulic acids showed similar concentrations lower than 5,mg,kg,1. The highest content of lycopene was found in Ramillete, Pera and Durina (>50,mg,kg,1 fresh weight), while the concentration in the other varieties was between 50 and 30,mg,kg,1, with the exception of Liso (less than 20,mg,kg,1). The antioxidant activity of tomato extracts varied with the tomato variety and the assay method used. Individual compounds found to be significantly related to antioxidant capacity were lycopene and ferulic and caffeic acids, but not quercetin and chlorogenic acid. © 2002 Society of Chemical Industry [source]

Bacterial and mammalian-cell genotoxicity of mixtures of chlorohydroxyfuranones, by-products of water chlorination

Jorma Mäki-Paakkanen
Abstract The genotoxic responses of mixtures of four chlorohydroxyfuranones (CHFs), 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX), 3,4-dichloro-5-hydroxy-2(5H)-furanone (MCA), 3-chloro- 4-(chloromethyl)-5-hydroxy-2(5H)-furanone (CMCF) and 3-chloro-4-methyl-5-hydroxy-2(5H)-furanone (MCF), were compared with the genotoxicity of the individual compounds. Genotoxicity was evaluated in the Salmonella reversion assay (Ames test), the in vitro Chinese hamster ovary (CHO) cell Hprt mutation assay, and in the CHO chromosome aberration test. When tested individually, the concentrations of the chemicals that were chosen for the mixtures induced no or only a modest increase in the genotoxic effects, and caused little or no cytotoxicity. In the Ames test, the genotoxic responses caused by the mixtures of CHFs did not follow simple additivity. Synergism was observed with strains TA97 and TA98, and antagonism with strain TA100. In the CHO/Hprt mutation assay, the mutagenic response of the mixtures was inconsistent, with near additivity seen with a mixture of CHFs that resulted in 12% cell survival. In contrast, the four CHFs together consistently caused more structural chromosome damage (mainly chromatid-type breaks and exchanges) compared to the sum of net effects of the four CHFs tested alone. Also, a potentiating effect was consistently seen for the cytotoxicity of the CHF mixtures both in the CHO/Hprt mutation assay and the chromosome aberration test. The present results indicate that the genotoxic effects of CHF mixtures can be greater than additive. Such effects may be worth considering in the cancer risk assessment of chlorinated drinking water. Environ. Mol. Mutagen. 43:217,225, 2004. © 2004 Wiley-Liss, Inc. [source]

Altered reproduction in fish exposed to pulp and paper mill effluents: Roles of individual compounds and mill operating conditions

L. Mark Hewitt
Abstract For the last 20 years, studies conducted in North America, Scandinavia, and New Zealand have shown that pulp and paper mill effluents affect fish reproduction. Despite the level of effort applied, few leads are available regarding the factors responsible. Effluents affect reproduction in multiple fish species, as evidenced by decreased gonad size, decreased circulating and gonadal production of reproductive steroids, altered expression of secondary sex characteristics, and decreased egg production. Several studies also have shown that effluent constituents are capable of accumulating in fish and binding to sex steroid receptors/binding proteins. Studies aimed at isolating biologically active substances within the pulping and papermaking process have provided clues about their source, and work has progressed in identifying opportunities for in-mill treatment technologies. Following comparisons of manufacturing processes and fish responses before and after process changes, it can be concluded that effluent from all types of mill processes are capable of affecting fish reproduction and that any improvements could not be attributed to a specific process modification (because mills normally performed multiple modifications simultaneously). Improved reproductive performance in fish generally was associated with reduced use of molecular chlorine, improved condensate handling, and liquor spill control. Effluent biotreatment has been effective in reducing some effects, but biotreated effluents also have shown no difference or an exacerbation of effects. The role of biotreatment in relation to effects on fish reproduction remains unclear and needs to be resolved. [source]

Can the choice of endpoint lead to contradictory results of mixture-toxicity experiments?

Nina Cedergreen
Abstract Theoretically, the effect of two independently acting compounds in a mixture will depend on the slope of the dose-response curves of the individual compounds if evaluated in relation to the model of concentration addition (CA). In the present study, we explored development of the shape of the dose-response relationship for four different recommended endpoints (surface area, frond number, fresh weight-specific, and dry weight-specific relative growth rates [RGRA, RGRF, RGRFW, and RGRDW, respectively]) and for two differently acting herbicides (metsulfuron-methyl and terbuthylazine) over time (3,15 d) on the standard test plant Lemna minor to identify endpoints and experiment times for which predictions of independent action (IA) would depart the most from those of CA. After a test time of 6 d, predictions of IA based on RGRA and RGRFW showed antagonism in relation to CA. Based on RGRDW, synergy was predicted, whereas IA based on RGRF was indistinguishable from CA. To test the prediction of choice of endpoint giving different results in mixture-toxicity experiments, three endpoints and six combinations of independently acting herbicides were evaluated using isobolograms. The experiments showed that in four of six herbicide combinations, different conclusions were reached depending on endpoint. The contradictory isoboles did not follow the theory of IA and, therefore, are more likely to be related to differences in susceptibility of the physiological processes affecting each endpoint than to the shape of the dose-response curve. [source]

Cytochrome P4501A induction potencies of polycyclic aromatic hydrocarbons in a fish hepatoma cell line: Demonstration of additive interactions

Karl Fent
Abstract The relative CYP1A induction potencies, determined as ethoxyresorufin- O -deethylase (EROD) activity, and the cytotoxicities of 19 compounds with one to six benzene rings, mixtures of polycyclic aromatic hydrocarbons (PAHs), and contaminated landfill leachates have been determined in the permanent fish hepatoma cell line PLHC-1. No CYP1A induction was observed with benzene, naphthalene, anthracene, acenaphthene, benzo[g,h,i]perylene, and fluorene and low induction was found with fluoranthene and phenanthrene. All other PAHs with three and more benzene rings led to a concentration-related induction of CYP1A, with rebound decreases at high concentrations resulting in bell-shaped concentration,activity curves. Fish-related induction equivalency factors (IEFs) were estimated for all PAHs on the basis of EC50 values of their EROD activities and are reported here for the first time. The following order of decreasing IEFs was found: dibenz[a,h]anthracene > dibenzo[a,i]pyrene > benzo[k]fluoranthene > 3-methylcholanthrene > benzo[a]pyrene > benzo[e]pyrene > chrysene > 7,12-dimethylbenz[a]anthracene > perylene > benz[a]anthracene > pyrene. In contrast to the EROD activity, the immunodetectable protein content determined by ELISA showed a concentration-dependent increase. The interaction of PAHs in mixtures of up to eight individual compounds was additive based on their EROD activities. In landfill leachates, determined induction equivalents (IEQ) were significantly higher than calculated IEQs based on analytical measurements, which indicates additional unknown inducing compounds present in leachates. This study shows that the PLHC-1 cell in vitro system serves as an integrative bioanalytical tool in the ecotoxicological evaluation of aquatic environmental samples contaminated with CYP1A-inducing compounds. [source]

Influence of carbonation on aroma release from liquid systems using an artificial throat and a proton transfer reaction,mass spectrometric technique (PTR,MS)

Maria Įngeles Pozo-Bayón
Abstract To determine whether carbonation affects aroma release from liquid systems, carbonated and non-carbonated flavoured model systems were prepared and volatile release was determined under static (equilibrium) and dynamic conditions. A model flavour system was added as a single compound or as a mixture of the six aroma compounds used in this study. Volatile release under dynamic conditions involved using a home-made device simulating an artificial throat, coupled to a proton transfer mass spectrometer (PTR,MS). The results showed that carbonation increased the release of most of the aroma compounds in both static and in dynamic testing conditions. The extent of this effect depended, however, on the physicochemical characteristics of the aroma compounds (the most volatile and most hydrophobic compounds were affected more). Release was also increased if the aroma compounds were added as a mixture rather than as individual compounds. CO2 appears to be a key factor responsible for the enhanced release of flavourings from carbonated liquid systems. Copyright © 2009 John Wiley & Sons, Ltd. [source]

Determination of linear response in the detection of mixtures of aroma compounds by atmospheric pressure ionization,mass spectrometry (API,MS)

Roberto A. Buffo
Abstract The linearity of atmospheric pressure ionization,mass spectrometry (API,MS) response in mixtures of aroma compounds was determined for a ,homogeneous' system (four esters) and a ,heterogeneous' system (five compounds of different chemical functionality). All combinations analysed within each system followed a saturation pattern as concentration increased. Total linear responses were determined by the linear behaviour of individual compounds. Ionization patterns within each mixture were explained by proton affinity data (i.e. the species' ability to accept protons), molecular reaction rates (defined by molecular weight), and product ion distribution (according to fragmentation of the corresponding parent ion). Copyright © 2007 John Wiley & Sons, Ltd. [source]

Phosphorylation of derivatives of ,-dialkyaminocrotonitriles with phosphorus(III) halides

Aleksandr N. Kostyuk
The reaction of ,-dialkylaminocrotontriles with phosphorus(III) halides has been investigated. The basicity of the dialkylamino group influences the phosphorylation markedly, with pyrrolidine being the amine of choice. It was found that a solvent and the ratio of triethylamine play a significant role in phosphorylation. Although chloro- and dichlorophosphine derivatives proved impossible to separate as individual compounds; their solutions can be successfully used for further transformations. © 2009 Wiley Periodicals, Inc. Heteroatom Chem 20:194,201, 2009; Published online in Wiley InterScience ( DOI 10.1002/hc.20532 [source]

Formation and decay of the ABTS derived radical cation: A comparison of different preparation procedures

Carola Henriquez
Bleaching of a preformed solution of the blue-green radical cation 2,2,-azinobis (3-ethylbenzothizoline-6-sulfonic acid) (ABTS+·) has been extensively used to evaluate the antioxidant capacity of complex mixtures and individual compounds. The reaction of the preformed radical with free-radical scavengers can be easily monitored by following the decay of the sample absorbance at 734 nm. The ABTS radical cation can be prepared employing different oxidants. Results obtained using MnO2 as oxidant show that the presence of manganese ions increases the rate of [ABTS]+· autobleaching in a concentration-dependent manner. The radicals can also be obtained by oxidizing ABTS with 2,2, -azobis(2-amidinopropane)hydrochloride (AAPH) or peroxodisulfate (PDS). The oxidation by AAPH takes place with a large activation energy and a low reaction order in ABTS. The data support a mechanism in which the homolysis of AAPH is the rate-limiting step, followed by the reaction of ABTS with the peroxyl radicals produced after the azocompound thermolysis. On the other hand, the low activation energy measured employing PDS, as well as the kinetic law, are compatible with the occurrence of a bimolecular reaction between the oxidant and ABTS. Regarding the use of ABTS-based methodologies for the evaluation of free radical scavengers, radical cations obtained employing AAPH as oxidant can be used only at low temperatures, conditions where further decomposition of the remaining AAPH is minimized. The best results are obtained with ABTS derived radicals generated in the reaction of PDS with an ABTS/PDS concentration ratio equal (or higher) to two. However, even with radicals prepared by this procedure, stoichiometric coefficients considerably larger than two are obtained for the consumption of the radical cation employing tryptophane or p -terbutylphenol as reductants. This casts doubts on the use of ABTS-based procedures for the estimation of antioxidant capacities. © 2002 Wiley Periodicals, Inc. Int J Chem Kinet 34: 659,665, 2002 [source]

Evaluation of semiochemicals potentially synergistic to ,-pinene for trapping the larger European pine shoot beetle, Tomicus piniperda (Col., Scolytidae)

T. M. Poland
Abstract:, The pine shoot beetle, Tomicus piniperda (L.) (Col., Scolytidae) is an exotic pest of pine, Pinus spp., in North America. It is attracted strongly to host volatiles (±)- , -pinene, (+)-3-carene, and , -terpinolene. Attraction to insect-produced compounds is less clear. Other potential attractants include trans -verbenol, myrtenol, myrtenal, nonanal and , -pinene oxide. We conducted a series of field experiments to determine if any of these compounds would increase attraction of T. piniperda to , -pinene, either individually or in various combinations. None of the individual compounds increased attraction. Although several combinations that included trans -verbenol, nonanal, myrtenol, or myrtenal increased attraction, results were variable between experiments. [source]

Research Article: Fingerprinting Analysis of Saposhnikovia divaricata using 1H Nuclear Magnetic Resonance Spectroscopy and High Performance Liquid Chromatography

Yue-Yang Xin
The 1H nuclear magnetic resonance (1H NMR) fingerprints of fractionated non-polar and polar extracts (control substance for plant drug [CSPD] A and B) from the roots of 12 specimens of Saposhnikovia divaricata (Turcz.) Schischk were achieved with Fourier Transform (FT)-NMR spectrometer and assigned by comparison to each other and to the 1H NMR spectra of the isolated individual compounds. These fingerprints were found to be uniform in terms of the specificity for the implication of all 12 specimens being systematically of the same origin. The uniformity was further affirmed by high performance liquid chromatography (HPLC), which also revealed exactly identical specificity for the identified S. divaricata species with the 1H NMR appearances of corresponding CSPD on the part of the composition of characteristic constituents when comparing to corresponding individual compounds. This investigation unambiguously shows that the specific signals from the chemotaxonomically significant compounds of chromones and coumarins in S. divaricata are exhibited distinctively in the composite features of both 1H NMR fingerprints and HPLC profiles. The 1H NMR and HPLC profiles established can successfully be used as reference for the authentication of the origin of S. divaricata species as well as for chemotaxonomic studies. [source]

Study of carvedilol by combined Raman spectroscopy and ab initio MO calculations

M. P. M. Marques
The novel cardioprotective drug carvedilol was studied by both Raman spectroscopy and ab initio molecular orbital methods (using the density functional theory approach). The spectra, acquired both for the solid samples and DMSO solutions as a function of pH, were assigned in view of the calculated wavenumbers and intensities, and also based on the experimental data obtained for individual compounds which comprise the molecule, namely carbazole and 1,2-dimethoxybenzene. The pH dependence of the Raman pattern of carvedilol was studied, and the pKa value of its secondary amine group was determined (pKa = 8.25) through pH titration experiments. This kind of information is of great significance for the understanding of the biochemical role of carvedilol, which is strongly determined by the acid,base behaviour of the molecule. Copyright © 2002 John Wiley & Sons, Ltd. [source]

In vitro studies on antioxidant activity of lignans isolated from sesame cake extract

Kizhiyedathu Polachira Suja
Abstract The antioxidant activity of compounds isolated from a methanolic extract of commercial sesame cake was studied using a peroxidation model and a radical-scavenging method. Pure compounds were isolated from the extract by preparative high-performance liquid chromatography (HPLC) and identified and confirmed as sesamol, sesamin, sesamolin, sesaminol diglucoside and sesaminol triglucoside by HPLC, infrared, nuclear magnetic resonance and mass spectrometry. When the rate of inhibition of lipid peroxidation and the superoxide radical-scavenging power of the individual compounds were evaluated, the compounds showed antioxidant activity to different extents. The antioxidant activity of compounds by the ,-carotene-bleaching assay followed the order sesamol > sesamolin , sesamin > butylated hydroxytoluene (BHT) > sesaminol triglucoside > sesaminol diglucoside. By the thiocyanate method the inhibition of linoleic acid peroxidation shown by sesamol, sesamin, sesamolin, sesaminol triglucoside, sesaminol diglucoside and BHT at 200 mg l,1 was 77, 60, 69, 32, 25 and 49% respectively. A concentration,dependent superoxide,scavenging effect was also shown by these compounds. Sesamolin had an appreciable effect at 300 and 500 mg l,1, while the other compounds were more effective at 100 mg l,1. The study also established the occurrence of sesamol in the methanolic extract of defatted sesame cake for the first time. Copyright © 2005 Society of Chemical Industry [source]

Web server suite for complex mixture analysis by covariance NMR

Fengli Zhang
Abstract Elucidation of the chemical composition of biological samples is a main focus of systems biology and metabolomics. Their comprehensive study requires reliable, efficient, and automatable methods to identify and quantify the underlying metabolites. Because nuclear magnetic resonance (NMR) spectroscopy is a rich source of molecular information, it has a unique potential for this task. Here we present a suite of public web servers (, termed COLMAR, which facilitates complex mixture analysis by NMR. The COLMAR web portal presently consists of three servers: COLMAR covariance calculates the covariance NMR spectrum from an NMR input dataset, such as a TOCSY spectrum; COLMAR DemixC method decomposes the 2D covariance TOCSY spectrum into a reduced set of nonredundant 1D cross sections or traces, which belong to individual mixture components; and COLMAR query screens the traces against a NMR spectral database to identify individual compounds. Examples are presented that illustrate the utility of this web server suite for complex mixture analysis. Copyright © 2009 John Wiley & Sons, Ltd. [source]

Solid-Phase Preparation of a Library Based on a Phenylalanine Scaffold

Aina Colombo
Abstract A convenient strategy (preliminary study, preproduction and production) for the solid-phase preparation of a library using 4-iodophenylalanine as a scaffold is described. The aromatic ring was first modified via the Suzuki reaction and the amino position was subsequently derivatized into amides, sulfonamides, amines, carbamates and ureas. The scope and limitations of all of the reactions carried out in parallel are discussed. The solid-phase synthesis of a library of 315 individual compounds was attempted by using seven boronic acids and nine representative compounds from each of the following classes: carboxylic acids, sulfonyl chlorides, aldehydes, alcohols and isocyanates. Owing to the failure of the amine derivatization, 297 compounds were ultimately obtained. [source]

Chemical ecology of obligate pollination mutualisms: testing the ,private channel' hypothesis in the Breynia,Epicephala association

Glenn P. Svensson
Summary ,Obligate mutualisms involving actively pollinating seed predators are among the most remarkable insect,plant relationships known, yet almost nothing is known about the chemistry of pollinator attraction in these systems. The extreme species specificity observed in these mutualisms may be maintained by specific chemical compounds through ,private channels'. Here, we tested this hypothesis using the monoecious Breynia vitis-idaea and its host-specific Epicephala pollinator as a model. ,Headspace samples were collected from both male and female flowers of the host. Gas chromatography with electroantennographic detection (GC-EAD), coupled gas chromatography,mass spectrometry, and olfactometer bioassays were used to identify the floral compounds acting as the pollinator attractant. ,Male and female flowers of B. vitis-idaea produced similar sets of general floral compounds, but in different ratios, and male flowers emitted significantly more scent than female flowers. A mixture of 2-phenylethyl alcohol and 2-phenylacetonitrile, the two most abundant compounds in male flowers, was as attractive to female moths as the male flower sample, although the individual compounds were slightly less attractive when tested separately. ,Data on the floral scent signals mediating obligate mutualisms involving active pollination are still very limited. We show that system-specific chemistry is not necessary for efficient host location by exclusive pollinators in these tightly coevolved mutualisms. [source]

Insecticidal, antifeedant and oviposition deterrent effects of the essential oil and individual compounds from leaves of Chloroxylon swietenia DC.

S Ravi Kiran
Abstract Essential oil from the leaves of Chloroxylon swietenia DC. was obtained by hydrodistillation and cold extraction, and the chemical composition was determined by gas chromatography and gas chromatography-mass spectrometry. The major identified components were limonene, geijerene, pregeijerene, germacrene D and trans-,-ocimene. Laboratory bioassays of the essential oil and four constituents of essential oil isolates were evaluated for insecticidal, antifeedant and oviposition deterrent effects on tobacco cutworm, Spodoptera litura (F.). Toxicity was determined by topical application of the isolates at varying concentrations. Pure oil, geijerene and pregeijerene were found to be more toxic, with LD50 values of 28.6, 35.4 and 40.7 µg per larva respectively. Maximum feeding deterrence was noted for geijerene and pregeijerene, with DC50 values of 82.5 and 95.1 µg cm,2 respectively. Furthermore, pure oil, geijerene and pregeijerene displayed oviposition deterrence, even at low concentrations. These results indicate that these natural products may lead to useful, biodegradable, environmentally safe insect control agents. Copyright © 2006 Society of Chemical Industry [source]

Insecticidal 2-hydroxy-3-alkyl-1,4-naphthoquinones: correlation of inhibition of ubiquinol cytochrome c oxidoreductase (complex III) with insecticidal activity

Philip J Jewess
Abstract The insecticidal and in vitro activities of four homologous series of 2-hydroxy and acetoxy-3-substituted-1,4-naphthoquinones have been measured and correlated with their (Log) octanol/water partition coefficients (Log Ko/w). In vitro activity against mitochondrial complex III was only exhibited by 2-hydroxy-3-alkyl-1,4-naphthoquinones, indicating that the 2-acetoxy compounds act as pro-insecticides. Good correlation was observed between in vivo activity against the two-spotted spider mite, Tetranychus urticae and inhibition of complex III isolated from blowfly flight muscle. Both hydroxy and acetoxy analogues of individual compounds exhibited similar levels of in vivo activity with optimum activity for analogues with Log Ko/w values of 7,8. In contrast, the acetoxy derivatives showed superior in vivo activity against the tobacco whitefly, Bemisia tabaci. Complex III isolated from whitefly was optimally inhibited by hydroxy analogues with lower Log Ko/w values (6.0,6.5) and was also more sensitive than the blowfly enzyme to all the compounds tested. © 2002 Society of Chemical Industry [source]

Determination of isokinetic ratios necessary for equimolar incorporation of carboxylic acids in the solid-phase synthesis of mixture-based combinatorial libraries

BIOPOLYMERS, Issue 1 2002
Achyuta N. Acharya
Abstract The methods used to study the relative reaction rates of 45 different aliphatic and aromatic carboxylic acids when coupled to resin-bound amino acid amides is described. Competition experiments involving the coupling of incoming carboxylic acids to resin-bound amino acid amides were performed. The relative composition of each N-acylated amino acid amide in the resulting mixtures was compared to controls prepared by physically mixing equal aliquots of individual compounds in order to study the relative reaction rates of the incoming carboxylic acids. The ratios of the incoming carboxylic acids were then iteratively adjusted to yield as close to equimolar products as possible. As expected, the steric and electronic nature of the incoming carboxylic acids was found to influence their relative reaction rates. The steric hindrance of the resin-bound amino acid appears to have a proportional effect on the reaction rates of the incoming carboxylic acids. N-acylated amino acid amides in the final mixtures, prepared using the final isokinetic ratios, were found to be approximately equimolar. © 2002 Wiley Periodicals, Inc. Biopolymers 65: 32,39, 2002 [source]

Transient Effects of Overexpressing Anthranilate Synthase , and , Subunits in Catharanthusroseus Hairy Roots

Christie A. M. Peebles
Catharanthus roseus produces two economically valuable anticancer drugs, vinblastine and vincristine. These drugs are members of the terpenoid indole alkaloids and accumulate in small quantities within the plant; thus these two drugs are expensive to produce. Metabolic engineering efforts have focused on increasing the alkaloids in this pathway through various means such as elicitation, precursor feeding, and gene overexpression. Recently we successfully expressed Arabidopsis genes encoding a feedback-insensitive anthranilate synthase , subunit under the control of the glucocorticoid-inducible promoter system and the anthranilate synthase , subunit under the control of a constitutive promoter in C. roseus hairy roots. In this work we look at the transient behaviors of terpenoid indole alkaloids over a 72 h induction period in late exponential growth phase cultures. Upon induction, the tryptophan, tryptamine, and ajmalicine pools accumulated over 72 h. In contrast, the lochnericine, hörhammericine, and tabersonine pools decreased and leveled out over the 72 h induction period. Visible changes within the individual compounds usually took from 4 to 12 h. [source]

Effect of Binary Combinations of Selected Toxic Compounds on Growth and Fermentation of Kluyveromyces marxianus

Jose M. Oliva
The inhibitory effects of various lignocellulose degradation products on glucose fermentation by the thermotolerant yeast Kluyveromycesmarxianus were studied in batch cultures. The toxicity of the aromatic alcohol catechol and two aromatic aldehydes (4-hydroxybenzaldehyde and vanillin) was investigated in binary combinations. The aldehyde furfural that usually is present in relatively high concentration in hydrolyzates from pentose degradation was also tested. Experiments were conducted by combining agents at concentrations that individually caused 25% inhibition of growth. Compared to the relative toxicity of the individual compounds, combinations of furfural with catechol and 4-hydroxybenzaldehyde were additive (50% inhibition of growth). The other binary combinations assayed (catechol with 4-hydroxybenzaldehyde, and vanillin with catechol, furfural, or 4-hydroxybenzaldehyde) showed synergistic effect on toxicity and caused a 60,90% decrease in cell mass production. The presence of aldehydes in the fermentation medium strongly inhibited cell growth and ethanol production. Kluyveromyces marxianusreduces aldehydes to their corresponding alcohols to mitigate the toxicity of these compounds. The total reduction of aldehydes was needed to start ethanol production. Vanillin, in binary combination, was dramatically toxic and was the only compound for which inhibition could not be overcome by yeast strain assimilation, causing a 90% reduction in both cell growth and fermentation. [source]

Biosynthesis of New Indigoid Inhibitors of Protein Kinases Using Recombinant Cytochrome P450 2A6

Zhongliu-Liu Wu
Glycogen synthase kinase-3 (GSK-3) is a potential drug target for a number of human diseases. Some indigoids have been found to be potent inhibitors of GSK-3, and individual compounds with better activity, specificity, and solubility are desired. In this work, a new disubstituted indigoid generation system was developed with a tryptophanase-deficient Escherichia coli strain as a host to express the human cytochrome P450 2A6 mutant L240C/N297Q, which catalyzes the oxidation of indole to isatin and indoxyl, which in turn react to generate indigoids. Forty-five substituted 1H -indoles from commercial sources were used as substrates in the system, and indigoid mixtures were tested as potential inhibitors of GSK-3. After preliminary screening, cell extracts with high inhibitory activity towards GSK-3,/, were fractionated, and the IC50 values of twelve individual indigoids were measured for GSK-3,/, as well as the protein kinases CDK1/cyclinB and CDK5/p25. Several indigoids, including an indigo, showed stronger inhibition than found in previous work. The most potent towards GSK-3,/,, dimethyl indirubin 5,5,-dicarboxylate (IC50 of 51,nM), was modified by chemical reactions. One product, indirubin 5,5,-dicarboxylic acid 5-methyl ester, inhibited GSK-3,/, with an IC50 of 14,nM and selectivity nearly 40-fold over CDK1 and CDK5. Indirubin-5-5,-dicarbonitrile was also modified to the corresponding 3,-oxime, which had low specificity but showed very high inhibition of all three kinases with IC50 values of 5, 13, and 10,nM towards GSK-3,/,, CDK1, and CDK5, respectively. Thus, this system has the potential to generate new indigoids with therapeutic potential. [source]