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Secondary Metabolism (secondary + metabolism)
Selected AbstractsModification of Primary and Secondary Metabolism of Potato Plants by Nitrogen Application Differentially Affects Resistance to Phytophthora infestans and Alternaria solaniPLANT BIOLOGY, Issue 5 2006K. Mittelstraß Abstract: Potato plants (Solanum tuberosum L. cv. Indira) were grown at two levels of N supply in the greenhouse. Plants supplied with 0.8 g N per plant (high N variant) showed significantly increased biomass as compared to plants without additional N fertilisation (low N variant). C/N ratio was lower and protein content was higher in leaves of the high N variant. The concentration of chlorogenic acids and flavonols was significantly lower in leaves from the high N variant. Whereas resistance to Alternaria solani increased when plants were supplied with additional nitrogen, these plants were more susceptible to Phytophthora infestans. After infection with both pathogens, we found a strong induction of p-coumaroylnoradrenaline and p-coumaroyloctopamine, which are identified for the first time in potato leaves and are discussed as resistance factors of other solanaceous plants. [source] Relationship Between Growth, Secondary Metabolism, and Resistance of ApplePLANT BIOLOGY, Issue 2 2002S. Rühmann Abstract: The paper shows that N-induced vigorous shoot growth increases susceptibility of apple trees to Venturia inaequalis. This is due to a weakened defence in infected leaves of the high N cultures showing large lesions with excessive sporulation, whereas infected leaves from the low N cultures exhibited successful defence with only small chlorotic lesions and no sporulation. This might be explained by biosynthesis of phenylpropanoids in the young leaves of the resistant trees. A negative correlation between shoot growth of apple trees and the concentration of phenolic compounds in young leaves was found. Studies on in vitro shoot cultures revealed that the availability of sugars for the phenylpropanoid pathway is a strong regulatory factor. The ratio of sucrose and nitrogen in the medium influenced the total level of secondary products in the in vitro grown plantlets. Moreover, the relative deficiency of sugars was responsible for a metabolic block mainly at the level of glucosyl transferase and concomitant aglycone accumulation. [source] The Auxiliary Substrate Concept: From simple considerations to heuristically valuable knowledgeENGINEERING IN LIFE SCIENCES (ELECTRONIC), Issue 4 2009Wolfgang Babel Abstract Microorganisms are used in biotechnology. They are either (i) aim and purpose of a process, e.g. with the production of single cell proteins, or (ii) mean to an end insofar as they serve as a catalyst or "factory" for syntheses (e.g. of products of primary and secondary metabolism, of enzymes and antibiotics) or for the degradation and detoxification of harmful organics and inorganics. In all cases, the efficiency and velocity, finally the productivity, are parameters which essentially determine the economy of the processes. Therefore, search for approaches to optimize these processes is a permanent task and challenge for scientists and engineers. It is shown that the auxiliary substrate concept is suitable to increase the yield coefficients. It is based on the energetic evaluation of organics, on the knowledge that organics as sources of carbon and energy for growth are deficient in ATP and/or reducing equivalents, and says that it is possible to improve the carbon conversion efficiency up to the carbon metabolism determined upper limit. The latter is determined by inevitable losses of carbon along the way of assimilation and anabolism and amounts to about 85% for so-called glycolytic substrates, e.g. glucose, methanol, and to about 75% for gluconeogenetic substrates, e.g. C2 -substrates (acetic acid, hexadecane). The approach is explained and some experimental examples are presented. By simultaneous utilization of an extra energy source (auxiliary substrate) the yield coefficient can be increased (i) in glucose from about 0.5 to 0.7,g/g (by means of formate), (ii) in acetate from 0.34,0.4 to 0.5,0.65,g/g (by means of formate and thiosulfate, respectively), and (iii) in hexadecane from about 0.94 to 1.26,g/g (by means of formate). The precalculated yield coefficients and mixing ratios agree well with the experimentally attained ones. The approach is easily feasible and economically valuable. [source] Molecular biology of aromatic plants and spices.FLAVOUR AND FRAGRANCE JOURNAL, Issue 5 2010A review. Abstract In recent years, molecular tools have been used to help to elucidate some aspects of genetic diversity in aromatic species, the genetic relationships between different cultivars and comparisons of molecular marker analysis to the chemical composition of plants. In this review, an explanation of the most important techniques involving molecular markers is given. A literature survey on molecular markers is presented, with some examples from aromatic plants and spices. However, understanding what controls flavour and aroma production in plants is not an easy task to accomplish. Several aspects of plant secondary metabolism, in particular volatiles production in aromatic plants, are still unknown. The route from genomics to proteomics is not well documented, although some research with model plants has already been performed. To address the question of the synthesis of volatiles, two different approaches are possible and summarized in this review: first, the biochemical and genetic approach; and second, approaches involving functional genomics. Finally, a brief survey of bioinformatics resources is presented. Copyright © 2010 John Wiley & Sons, Ltd. [source] Effects of ultrasound on culture of Aspergillus terreusJOURNAL OF CHEMICAL TECHNOLOGY & BIOTECHNOLOGY, Issue 5 2008Nuria Sainz Herrán Abstract BACKGROUND: Fermentations of Aspergillus terreus are commercially used to produce lovastatin. How ultrasound might influence this fermentation is unknown. While high-intensity ultrasound is effective in disrupting microbial cells, ultrasound of low intensity is known to improve productivity of some fermentation processes without damaging cells. Mechanisms behind productivity improvements have not been clearly identified in earlier studies. This work reports on the effects of ultrasound on A. terreus fermentation for low (957 W m,3), medium (2870 W m,3) and high (4783 W m,3) values of sonication power input in a slurry bubble column sonobioreactor. RESULTS: Sonication at any power level did not affect biomass growth profiles in comparison with negative controls. In contrast, medium- and high-intensity sonication greatly reduced production of lovastatin and substantially altered the growth morphology. At medium and high intensity, ultrasound disrupted fungal pellets and caused the biomass to grow mainly as dispersed hyphae. Sonication affected broth rheology because rheology depends on the morphology of the suspended biomass. CONCLUSION: Sonication can be used to modify growth morphology and broth rheology without affecting growth of filamentous fungi. Sonication appears to influence the primary growth metabolism and secondary metabolism differently in different situations. Copyright © 2008 Society of Chemical Industry [source] Biocontrol and Plant Pathogenic Fusarium oxysporum -Induced Changes in Phenolic Compounds in Tomato Leaves and RootsJOURNAL OF PHYTOPATHOLOGY, Issue 7-8 2007Y. Panina Abstract The biocontrol fungus Fusarium oxysporum strain CS-20 was previously shown to reduce the incidence of Fusarium wilt of tomato through an uncharacterized host-mediated response. As phenolic compounds are involved in the defence response of tomato to pathogens and other stressors, this work was undertaken to determine whether biocontrol strains induced changes in phenolic compounds in leaves and roots of tomato seedlings in the presence and absence of pathogenic F. oxysporum f. sp. lycopersici. Roots of intact tomato seedlings were placed in water or aqueous fungal spore suspensions. Two biocontrol F. oxysporum strains [CS-20 (host-mediated mechanism) and 85SK-1 (control mechanism unknown)] and two plant pathogenic strains of F. oxysporum f. sp. lycopersici Race 1 were used. After 24 or 72 h exposure, phenolic compounds were extracted from leaves and roots before identification by HPLC. There were significant qualitative and quantitative differences between the two sampling times. Compared with the control treatment, strain CS-20 significantly altered (usually increasing) the ferulic, caffeic and vanillic acid contents, and concentrations once unidentified phenolic compounds recovered from leaves and roots. In another experiment, tomato seedlings growing in sterile sand were drenched with spores of strain CS-20 the day before treating them with varying concentrations of spores of the pathogen for 24 or 72 h. The amount of pathogen present did not significantly affect the plant phenolic response to the presence of strain CS-20. This work demonstrates that tomato responds within 24 h to the presence of the biocontrol strain CS-20 by alterations in secondary metabolism that are typical of resistance responses in tomato. [source] Accumulation of Mycosporine-like Amino Acids in Asparagopsis armata Grown in Tanks with Fishpond Effluents of Gilthead Sea Bream, Sparus aurataJOURNAL OF THE WORLD AQUACULTURE SOCIETY, Issue 5 2008Félix L. Figueroa Both the effects of total ammonium nitrogen (TAN) fluxes and the algal densities on MAA accumulation were investigated. MAAs increased with the TAN flux, but only until values lower than 100 ,M/h. Above this flux, the MAA content decreased, whereas algal yield increased. The content of individual MAAs was related to nitrogen (N) status, that is, shinorine percentage slightly decreased and palythine increased with increasing N fluxes. The decrease of MAAs at high flux of N (up to 100 ,M/h) is related to the decrease of water residence time and the decrease of the biofiltration efficiency. Under different TAN fluxes and algal densities, MAA content was negatively related to algal yield indicating that MAAs were accumulated only under a high ammonium-N availability. Thus, an energy allocation between growth (primary metabolism) and MAA accumulation (secondary metabolism) is regulated by the absorption capacity of inorganic N. In conclusion, A. armata, in addition to its high biofiltration capacity of nutrients, is a good source of MAAs as potential UV screen photoprotectors. [source] CPCR1, but not its interacting transcription factor AcFKH1, controls fungal arthrospore formation in Acremonium chrysogenumMOLECULAR MICROBIOLOGY, Issue 5 2005Birgit Hoff Summary Fungal morphogenesis and secondary metabolism are frequently associated; however, the molecular determinants connecting both processes remain largely undefined. Here we demonstrate that CPCR1 (cephalosporin C regulator 1 from Acremonium chrysogenum), a member of the winged helix/regulator factor X (RFX) transcription factor family that regulates cephalosporin C biosynthesis, also controls morphological development in the ,-lactam producer A. chrysogenum. The use of a disruption strain, multicopy strains as well as several recombinant control strains revealed that CPCR1 is required for hyphal fragmentation, and thus the formation of arthrospores. In a ,cpcR1 disruption strain that exhibits only hyphal growth, the wild-type cpcR1 gene was able to restore arthrospore formation; a phenomenon not observed for ,cpcR1 derivatives or non-related genes. The intracellular expression of cpcR1, and control genes (pcbC, egfp) was determined by in vivo monitoring of fluorescent protein fusions. Further, the role of the forkhead transcription factor AcFKH1, which directly interacts with CPCR1, was studied by generating an Acfkh1 knockout strain. In contrast to CPCR1, AcFKH1 is not directly involved in the fragmentation of hyphae. Instead, the presence of AcFKH1 seems to be necessary for CPCR1 function in A. chrysogenum morphogenesis, as overexpression of a functional cpcR1 gene in a ,Acfkh1 background has no effect on arthrospore formation. Moreover, strains lacking Acfkh1 exhibit defects in cell separation, indicating an involvement of the forkhead transcription factor in mycelial growth of A. chrysogenum. Our data offer the potential to control fungal growth in biotechnical processes that require defined morphological stages for optimal production yields. [source] amfR, an essential gene for aerial mycelium formation, is a member of the AdpA regulon in the A-factor regulatory cascade in Streptomyces griseusMOLECULAR MICROBIOLOGY, Issue 4 2003Haruka Yamazaki Summary In Streptomyces griseus, A-factor (2-isocapryloyl-3R -hydroxymethyl-,-butyrolactone) acts as a chemical signalling molecule that triggers morphological differentiation and secondary metabolism. A transcriptional activator, AdpA, in the A-factor regulatory cascade switches on a number of genes required for both processes, thus forming an AdpA regulon. amfR encoding a regulatory protein similar to response regulators of bacterial two-component regulatory systems and essential for aerial mycelium formation was found to be a member of the AdpA regulon. AdpA bound two sites at nucleotide positions approximately ,200 (site 1) and ,60 (site 2), with respect to the major transcriptional start point of amfR, and accelerated the transcription of amfR by assisting RNA polymerase in forming an open complex at an appropriate region including the transcriptional start point. Site 2 contributed more to the transcriptional activation of amfR by AdpA than site 1, although AdpA showed a much lower affinity to site 2 than to site 1. The amfR transcription enhanced by AdpA subsequently ceased at day 2 when aerial hyphae began to be formed in the wild-type strain, whereas in an adsA null mutant amfR was continuously transcribed even until day 3. This implied that amfR was repressed growth dependently by a gene product under the control of ,-AdsA. Transcription of the promoter upstream of amfT depended on amfR, which is consistent with the idea that AmfR serves as an activator for amfTSBA in the amf operon. The observations that the amfR gene contains a TTA codon, a potential target for bldA -mediated regulation, and a conserved Asp-54 residue, which might be phosphorylated by a sensor kinase, suggest that the amf operon is under transcriptional, translational and post-translational control systems. [source] Primary and secondary metabolism, and post-translational protein modifications, as portrayed by proteomic analysis of Streptomyces coelicolorMOLECULAR MICROBIOLOGY, Issue 4 2002A. R. Hesketh Summary The newly sequenced genome of Streptomyces coelicolor is estimated to encode 7825 theoretical proteins. We have mapped approximately 10% of the theoretical proteome experimentally using two-dimensional gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry. Products from 770 different genes were identified, and the types of proteins represented are discussed in terms of their anno-tated functional classes. An average of 1.2 proteins per gene was observed, indicating extensive post-translational regulation. Examples of modification by N-acetylation, adenylylation and proteolytic processing were characterized using mass spectrometry. Proteins from both primary and certain secondary metabolic pathways are strongly represented on the map, and a number of these enzymes were identified at more than one two-dimensional gel location. Post-translational modification mechanisms may therefore play a significant role in the regulation of these pathways. Unexpectedly, one of the enzymes for synthesis of the actinorhodin polyketide antibiotic appears to be located outside the cytoplasmic compartment, within the cell wall matrix. Of 20 gene clusters encoding enzymes characteristic of secondary metabolism, eight are represented on the proteome map, including three that specify the production of novel metabolites. This information will be valuable in the characterization of the new metabolites. [source] Requirement of spermidine for developmental transitions in Aspergillus nidulansMOLECULAR MICROBIOLOGY, Issue 3 2002Yuan Jin Summary Deletion of the spermidine synthase gene in the fungus Aspergillus nidulans results in a strain, ,spdA, which requires spermidine for growth and accumulates putrescine as the sole polyamine. Vegetative growth but not sporulation or sterigmatocystin production is observed when ,spdA is grown on media supplemented with 0.05,0.10 mM exogenous spermidine. Supplementation of ,spdA with , 0.10 mM spermidine restores sterigmatocystin production and , 0.50 mM spermidine produces a phenotype with denser asexual spore production and decreased radial hyphal growth compared with the wild type. ,spdA spores germinate in unsupplemented media but germ tube growth ceases after 8 h upon which time the spores swell to approximately three times their normal diameter. Hyphal growth is resumed upon addition of 1.0 mM spermidine. Suppression of a G protein signalling pathway could not force asexual sporulation and sterigmatocystin production in ,spdA strains grown in media lacking spermidine but could force both processes in ,spdA strains supplemented with 0.05 mM spermidine. These results show that increasing levels of spermidine are required for the transitions from (i) germ tube to hyphal growth and (ii) hyphal growth to tissue differentiation and secondary metabolism. Suppression of G protein signalling can over-ride the spermidine requirement for the latter but not the former transition. [source] The evolution of secondary metabolism , a unifying modelMOLECULAR MICROBIOLOGY, Issue 5 2000Richard D. Firn Why do microbes make secondary products? That question has been the subject of intense debate for many decades. There are two extreme opinions. Some argue that most secondary metabolites play no role in increasing the fitness of an organism. The opposite view, now widely held, is that every secondary metabolite is made because it possesses (or did possess at some stage in evolution) a biological activity that endows the producer with increased fitness. These opposing views can be reconciled by recognizing that, because of the principles governing molecular interactions, potent biological activity is a rare property for any molecule to possess. Consequently, in order for an organism to evolve the rare potent, biologically active molecule, a great many chemical structures have to be generated, most of which will possess no useful biological activity. Thus, the two sides of the debate about the role and evolution of secondary metabolism can be accommodated within the view that the possession of secondary metabolism can enhance fitness, but that many products of secondary metabolism will not enhance the fitness of the producer. It is proposed that secondary metabolism will have evolved such that traits that optimize the production and retention of chemical diversity at minimum cost will have been selected. Evidence exists for some of these predicted traits. Opportunities now exist to exploit these unique properties of secondary metabolism to enhance secondary product diversity and to devise new strategies for biotransformation and bioremediation. [source] Gene duplications and the time thereafter , examples from plant secondary metabolismPLANT BIOLOGY, Issue 4 2010D. Ober Abstract Gene duplications are regarded as one of the central mechanisms for the origin of new genes. Recent studies in plant secondary metabolism have provided several examples of genes that originated by duplication with successive diversification. In this review, the mechanisms of gene duplication are explained and several models discussed that suggest the way that gene duplicates develop into genes with new functions. Signatures of gene duplication and diversification processes are discussed using the biosynthesis of benzoxazinones and of pyrrolizidine alkaloids as examples. [source] Molecular Analysis of Brassinosteroid ActionPLANT BIOLOGY, Issue 3 2006C. Müssig Abstract: Brassinosteroids (BRs) are steroidal plant hormones with important regulatory roles in various physiological processes, including growth, xylem differentiation, disease resistance, and stress tolerance. Several components of the BR signal transduction pathway have been identified. The extracellular domains of receptor kinases such as BRI1 perceive BRs and transduce the signal via intracellular kinase domains. Within the cell further kinases and phosphatases determine the phosphorylation status of transcription factors such as BES1 and BZR1. These factors mediate major BR effects. Studies of BR-regulated genes shed light on the molecular mode of BR action. Genes encoding cell-wall-modifying enzymes, enzymes of the BR biosynthetic pathway, transcription factors, and proteins involved in primary and secondary metabolism are subject to BR-regulation. Gene expression data also point at interactions with other phytohormones and a role of BR in stress responses. This article gives a survey of the BR-signaling pathway. Two BR-responsive genes, OPR3 and EXO, are described in detail. [source] Significance of Flavonoids in Plant Resistance and Enhancement of Their BiosynthesisPLANT BIOLOGY, Issue 6 2005D. Treutter Abstract: The roles of flavonoids in plant defence against pathogens, herbivores, and environmental stress are reviewed and their significant contribution to plant resistance is discussed. The induction of flavonoids is of particular interest for gathering evidence of their roles. Tools are mentioned which may enhance flavonoid biosynthesis and accumulation. These include metabolic engineering and UV light. The induction of defence-related flavonoids is modified by other determining factors and competition between growth and secondary metabolism may exist. In an evolutionary context, stress-related oxidative pressure may have been a major trigger for the distribution and abundance of flavonoids. UV protection is one of their most significant, or even the most significant, functional role for flavonoids. The multi-functionality of these compounds, however, often complicates the interpretation of experimental results but, overall, it supports the importance of flavonoids. [source] Growth-Promoting Nitrogen Nutrition Affects Flavonoid Biosynthesis in Young Apple (Malus domestica Borkh.) LeavesPLANT BIOLOGY, Issue 6 2005T. Strissel Abstract: Enhanced shoot growth and a decrease in flavonoid concentration in apple trees grown under high nitrogen (N) supply was observed in previous studies, along with increasing scab susceptibility of cultivar "Golden Delicious" after high N nutrition. Several hypotheses have suggested that there is a trade-off between primary and secondary metabolism because of competition for common substrates, but nothing is known about regulation at the enzyme level. In this study, a set of experiments was performed to elucidate the effect of N nutrition on the activities of key enzymes involved in flavonoid biosynthesis (phenylalanine ammonia-lyase [PAL], chalcone synthase/chalcone isomerase [CHS/CHI}, flavanone 3-hydroxylase [FHT], flavonol synthase [FLS], dihydroflavonol 4-reductase [DFR]) and the accumulation of different groups of phenylpropanoids. The inhibition of flavonoid accumulation by high N nutrition could be confirmed, but the influence of N supply on the flavonoid enzymes CHS/CHI, FHT, DFR, and FLS was not evident. However, PAL activity seems to be downregulated, thus forming a bottleneck resulting in a generally decreased flavonoid accumulation. Furthermore, the response of the scab-resistant cultivar "Rewena" to high N nutrition was not as strong as that of the susceptible cultivar "Golden Delicious". [source] Carbon metabolite sensing and signallingPLANT BIOTECHNOLOGY JOURNAL, Issue 6 2003Nigel G. Halford Abstract The regulation of carbon metabolism in plant cells responds sensitively to the levels of carbon metabolites that are available. The sensing and signalling systems that are involved in this process form a complex web that comprises metabolites, transporters, enzymes, transcription factors and hormones. Exactly which metabolites are sensed is not yet known, but candidates include sucrose, glucose and other hexoses, glucose-6-phosphate, trehalose-6-phosphate, trehalose and adenosine monophosphate. Important components of the signalling pathways include sucrose non-fermenting-1-related protein kinase-1 (SnRK1) and hexokinase; sugar transporters are also implicated. A battery of genes and enzymes involved in carbohydrate metabolism, secondary metabolism, nitrogen assimilation and photosynthesis are under the control of these pathways and fundamental developmental processes such as germination, sprouting, pollen development and senescence are affected by them. Here we review the current knowledge of carbon metabolite sensing and signalling in plants, drawing comparisons with homologous and analogous systems in animals and fungi. We also review the evidence for cross-talk between carbon metabolite and other major signalling systems in plant cells and the prospects for manipulating this fundamentally important aspect of metabolic regulation for crop improvement. [source] Proteomic evaluation of wound-healing processes in potato (Solanum tuberosum L.) tuber tissuePROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 17 2009Inês Chaves Abstract Proteins from potato (Solanum tuberosum L.) tuber slices, related to the wound-healing process, were separated by 2-DE and identified by an MS analysis in MS and MS/MS mode. Slicing triggered differentiation processes that lead to changes in metabolism, activation of defence and cell-wall reinforcement. Proteins related to storage, cell growth and division, cell structure, signal transduction, energy production, disease/defence mechanisms and secondary metabolism were detected. Image analysis of the 2-DE gels revealed a time-dependent change in the complexity of the polypeptide patterns. By microscopic observation the polyalyphatic domain of suberin was clearly visible by D4, indicating that a closing layer (primary suberisation) was formed by then. A PCA of the six sampling dates revealed two time phases, D0,D2 and D4,D8, with a border position between D2 and D4. Moreover, a PCA of differentially expressed proteins indicated the existence of a succession of proteomic events leading to wound-periderm reconstruction. Some late-expressed proteins (D6,D8), including a suberisation-associated anionic peroxidase, have also been identified in the native periderm. Despite this, protein patterns of D8 slices and native periderm were still different, suggesting that the processes of wound-periderm formation are extended in time and not fully equivalent. The information presented in this study gives clues for further work on wound healing-periderm formation processes. [source] Reprogramming a maize plant: transcriptional and metabolic changes induced by the fungal biotroph Ustilago maydisTHE PLANT JOURNAL, Issue 2 2008Gunther Doehlemann Summary The fungal pathogen Ustilago maydis establishes a biotrophic relationship with its host plant maize (Zea mays). Hallmarks of the disease are large plant tumours in which fungal proliferation occurs. Previous studies suggested that classical defence pathways are not activated. Confocal microscopy, global expression profiling and metabolic profiling now shows that U. maydis is recognized early and triggers defence responses. Many of these early response genes are downregulated at later time points, whereas several genes associated with suppression of cell death are induced. The interplay between fungus and host involves changes in hormone signalling, induction of antioxidant and secondary metabolism, as well as the prevention of source leaf establishment. Our data provide novel insights into the complexity of a biotrophic interaction. [source] The multiple phenylpropene synthases in both Clarkia breweri and Petunia hybrida represent two distinct protein lineagesTHE PLANT JOURNAL, Issue 3 2008Takao Koeduka Summary Many plants synthesize the volatile phenylpropene compounds eugenol and isoeugenol to serve in defense against herbivores and pathogens and to attract pollinators. Clarkia breweri flowers emit a mixture of eugenol and isoeugenol, while Petunia hybrida flowers emit mostly isoeugenol with small amounts of eugenol. We recently reported the identification of a petunia enzyme, isoeugenol synthase 1 (PhIGS1) that catalyzes the formation of isoeugenol, and an Ocimum basilicum (basil) enzyme, eugenol synthase 1 (ObEGS1), that produces eugenol. ObEGS1 and PhIGS1 both utilize coniferyl acetate, are 52% sequence identical, and belong to a family of NADPH-dependent reductases involved in secondary metabolism. Here we show that C. breweri flowers have two closely related proteins (96% identity), CbIGS1 and CbEGS1, that are similar to ObEGS1 (58% and 59% identity, respectively) and catalyze the formation of isoeugenol and eugenol, respectively. In vitro mutagenesis experiments demonstrate that substitution of only a single residue can substantially affect the product specificity of these enzymes. A third C. breweri enzyme identified, CbEGS2, also catalyzes the formation of eugenol from coniferyl acetate and is only 46% identical to CbIGS1 and CbEGS1 but more similar (>70%) to other types of reductases. We also found that petunia flowers contain an enzyme, PhEGS1, that is highly similar to CbEGS2 (82% identity) and that converts coniferyl acetate to eugenol. Our results indicate that plant enzymes with EGS and IGS activities have arisen multiple times and in different protein lineages. [source] Regulation of secondary metabolism by the carbon,nitrogen status in tobacco: nitrate inhibits large sectors of phenylpropanoid metabolismTHE PLANT JOURNAL, Issue 4 2006Christina Fritz Summary Interactions between nitrogen and carbon metabolism modulate many aspects of the metabolism, physiology and development of plants. This paper investigates the contribution of nitrate and nitrogen metabolism to the regulation of phenylpropanoid and nicotine synthesis. Wild-type tobacco was grown on 12 or 0.2 mm nitrate and compared with a nitrate reductase-deficient mutant [Nia30(145)] growing on 12 mm nitrate. Nitrate-deficient wild-type plants accumulate high levels of a range of phenylpropanoids including chlorogenic acid, contain high levels of rutin, are highly lignified, but contain less nicotine than nitrogen-replete wild-type tobacco. Nia30(145) resembles nitrate-deficient wild-type plants with respect to the levels of amino acids, but accumulates large amounts of nitrate. The levels of phenylpropanoids, rutin and lignin resemble those in nitrogen-replete wild-type plants, whereas the level of nicotine resembles that in nitrate-deficient wild-type plants. Expression arrays and real time RT-PCR revealed that a set of genes required for phenylpropanoid metabolism including PAL, 4CL and HQT are induced in nitrogen-deficient wild-type plants but not in Nia30(145). It is concluded that nitrogen deficiency leads to a marked shift from the nitrogen-containing alkaloid nicotine to carbon-rich phenylpropanoids. The stimulation of phenylpropanoid metabolism is triggered by changes of nitrate, rather than downstream nitrogen metabolites, and is mediated by induction of a set of enzymes in the early steps of the phenylpropanoid biosynthetic pathway. [source] New insights into plant transaldolaseTHE PLANT JOURNAL, Issue 1 2005Maxime Caillau Summary The oxidative pentose phosphate pathway (OPPP) provides plants with important substrates for both primary and secondary metabolism via the oxidation of glucose-6-phosphate. The OPPP is also thought to generate large amounts of reducing power to drive various anabolic processes. In animals this major pathway is located within the cytoplasm of cells, but in plants its subcellular compartmentation is far from clear. Although several enzymes of the OPPP were demonstrated to have both cytosolic and plastidic counterparts, there is yet no evidence for a full set of functional enzymes in each compartment. We report here the isolation of two coding sequences from tomato (Lycopersicon esculentum L.) which encode phylogenetically distant sequences (ToTal1 and ToTal2) that putatively encode distinct plastidic TA isoforms. The kinetic characterization of ToTal1 revealed that, unlike other enzymes of the non-oxidative branch of the OPPP, ToTal1 does not follow a Michaelis,Menten mode of catalysis which has implications for its role in regulating carbon flux between primary and secondary metabolism. TA genes appear to be differentially regulated at the level of gene expression in plant tissues and in response to environmental factors which suggests that TA isoforms have a non-overlapping role for plant metabolism. [source] Identification and characterization of a novel anthocyanin malonyltransferase from scarlet sage (Salvia splendens) flowers: an enzyme that is phylogenetically separated from other anthocyanin acyltransferasesTHE PLANT JOURNAL, Issue 6 2004Hirokazu Suzuki Summary Anthocyanin acyltransferases (AATs) catalyze a regiospecific acyl transfer from acyl-CoA to the glycosyl moiety of anthocyanins, thus playing an important role in flower coloration. The known AATs are subfamily members of an acyltransferase family, the BAHD family, which play important roles in secondary metabolism in plants. Here, we describe the purification, characterization, and cDNA cloning of a novel anthocyanin malonyltransferase from scarlet sage (Salvia splendens) flowers. The purified enzyme (hereafter referred to as Ss5MaT2) is a monomeric 46-kDa protein that catalyzes the transfer of the malonyl group from malonyl-CoA to the 4,,,-hydroxyl group of the 5-glucosyl moiety of anthocyanins. Thus, it is a malonyl-CoA:anthocyanin 5-glucoside 4,,,- O -malonyltransferase. On the basis of the partial amino acid sequences of the purified enzyme, we isolated a cDNA that encodes an acyltransferase protein. The steady-state transcript level of the gene was the highest in recently opened, fully pigmented flowers and was also correlated with the trend observed for an AAT gene responsible for the first malonylation step during salvianin biosynthesis. Immunoprecipitation studies using antibodies against the recombinant acyltransferase protein corroborated the identity of this cDNA as that encoding Ss5MaT2. The deduced amino acid sequence of Ss5MaT2 showed a low similarity (22,24% identity) to those of AATs and lacked the AAT-specific signature sequence. A phylogenetic analysis suggested that Ss5MaT2 is more related to acetyl-CoA:benzylalcohol acetyltransferase (BEAT) rather than to AAT. This is another example in which enzymes with similar, although not identical, substrate evolved from different branches of the BAHD family. [source] Effect of the plant peptide regulator, phytosulfokine-,, on the growth and Taxol production from Taxus sp. suspension culturesBIOTECHNOLOGY & BIOENGINEERING, Issue 1 2006Beum Jun Kim Abstract Phytosulfokine-, (PSK-,) is a small plant peptide (5 amino acids) that displays characteristics typically associated with animal peptide hormones. PSK-, was originally isolated based on its mitogenic activity with plant cultures; it has been reported to increase production of tropane alkaloids from Atropa belladonna, although its general influence on secondary metabolite production is unknown. The studies reported in this article were initiated to evaluate the effects of PSK-, supplementation on production of TaxolÔ (paclitaxel) from plant cell cultures of Taxus sp. particularly when methyl jasmonate (MeJA) is added as an elicitor of secondary metabolism. The response to PSK-, supplementation was cell line dependent. Taxus cuspidata P93AF showed no statistically significant response to PSK-, supplementation while Taxus canadensis C93AD and T. cuspidata PO93X displayed a concentration-dependent response (up to 100 nM PSK-, added in first 24 h of culture) with a decrease in initial growth rate, an increase in cell density (dry weight/fresh weight), and increased Taxol production. More remarkably with T. canadensis (C93AD), a very strong synergistic response of PSK-, (100 nM) and methyl jasmonate (MeJA, 100 µM) elicitation was observed, resulting in Taxol level of 35.3,±,2.1 mg/L or 1.83,±,0.02 mg Taxol/g dry cell weight achieved at day 21, a level of approximately 10-fold higher than for either treatment by itself. Although the level of Taxol production achieved is not remarkable, this synergistic treatment was able to partially revive taxane production in cultures that have lost productivity due to extended time (over 10 years) in continuous subculture. © 2006 Wiley Periodicals, Inc. [source] Invariability of central metabolic flux distribution in Shewanella oneidensis MR-1 under environmental or genetic perturbationsBIOTECHNOLOGY PROGRESS, Issue 5 2009Yinjie J. Tang Abstract An environmentally important bacterium with versatile respiration, Shewanella oneidensis MR-1, displayed significantly different growth rates under three culture conditions: minimal medium (doubling time ,3 h), salt stressed minimal medium (doubling time ,6 h), and minimal medium with amino acid supplementation (doubling time ,1.5 h). 13C-based metabolic flux analysis indicated that fluxes of central metabolic reactions remained relatively constant under the three growth conditions, which is in stark contrast to the reported significant changes in the transcript and metabolite profiles under various growth conditions. Furthermore, 10 transposon mutants of S. oneidensis MR-1 were randomly chosen from a transposon library and their flux distributions through central metabolic pathways were revealed to be identical, even though such mutational processes altered the secondary metabolism, for example, glycine and C1 (5,10-Me-THF) metabolism. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 [source] Phenylnannolones A,C: Biosynthesis of New Secondary Metabolites from the Myxobacterium Nannocystis exedensCHEMBIOCHEM, Issue 18 2008Birgit Ohlendorf Abstract Myxobacteria are gliding bacteria that belong to the ,-Proteobacteria and are known for their unique biosynthetic capabilities. Among myxobacteria, Nannocystis spp. are most closely related to marine myxobacteria and their secondary metabolism has hardly been investigated. Phenylnannolones A (1), B (2) and C (3) were obtained from a culture of Nannocystis exedens that was isolated from the intertidal region of Crete. Compound 1 had inhibitory activity toward the ABCB1 gene product P-glycoprotein and reversed daunorubicin resistance in cultured cancer cells. Phenylnannolone A has an unusual structural architecture; it is composed of an ethyl-substituted polyene chain linked to a pyrone moiety on one side and to a phenyl ring on the other. The investigation of the biosynthesis with labelled precursors revealed acetate, butyrate and phenylalanine as building blocks for 1. The labelling pattern suggested novel biochemical reactions for the biosynthesis of the starter unit. [source] Analysis of intracellular short organic acid-coenzyme A esters from actinomycetes using liquid chromatography-electrospray ionization-mass spectrometryJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 9 2007Je Won Park Abstract A method employing silicone oil density centrifugation, solid-phase extraction (SPE) cleanup, and LC-ESI-MS/MS analysis was developed for the rapid, selective, sensitive, and quantitative detection of an intracellular pool of short organic acid-CoA esters in actinomycetes. The detection limit was determined to be approximately 0.8 pmol (1.2 ng/ml) for each standard CoA-ester analyzed by the present LC-ESI-MS/MS method. A selected ion chromatogram for a typical fragment ion (m/z 428) specific to CoA-esters enabled the detection of eight intracellular CoA-esters involved in both primary and secondary metabolisms. The application of this method to bacterial metabolomic study is demonstrated by the profiling of the intracellular CoA-ester pools in the wild-type Streptomyces venezuelae strain producing polyketide antibiotics (methymycin and pikromycin), a polyketide synthase (PKS)-deleted S. venezuelae mutant, and a S. venezuelae mutant expressing the heterologous PKS genes. By quantifying the individual CoA-esterlevel in three different genotypes of the S. venezuela e strain, further insight could be gained into the role of CoA-estersin polyketide biosynthesis. This analytical approach can be extended to the quantification of the size and composition of in vivo CoA-ester pools in various microbes, and can provide a detailed understanding of the relationship between the in vivo CoA-ester pool and the production of pharmaceutically important polyketides. Copyright © 2007 John Wiley & Sons, Ltd. [source] | |||||||||||||||||||||||||