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Phenylpropanoid Pathway (phenylpropanoid + pathway)
Selected AbstractsMetabolic engineering of Saccharomyces cerevisiae for the synthesis of the wine-related antioxidant resveratrolFEMS YEAST RESEARCH, Issue 1 2003John V.W. Becker Abstract The stilbene resveratrol is a stress metabolite produced by Vitis vinifera grapevines during fungal infection, wounding or UV radiation. Resveratrol is synthesised particularly in the skins of grape berries and only trace amounts are present in the fruit flesh. Red wine contains a much higher resveratrol concentration than white wine, due to skin contact during fermentation. Apart from its antifungal characteristics, resveratrol has also been shown to have cancer chemopreventive activity and to reduce the risk of coronary heart disease. It acts as an antioxidant and anti-mutagen and has the ability to induce specific enzymes that metabolise carcinogenic substances. The objective of this pilot study was to investigate the feasibility of developing wine yeasts with the ability to produce resveratrol during fermentation in both red and white wines, thereby increasing the wholesomeness of the product. To achieve this goal, the phenylpropanoid pathway in Saccharomyces cerevisiae would have to be introduced to produce p -coumaroyl-CoA, one of the substrates required for resveratrol synthesis. The other substrate for resveratrol synthase, malonyl-CoA, is already found in yeast and is involved in de novo fatty-acid biosynthesis. We hypothesised that production of p -coumaroyl-CoA and resveratrol can be achieved by co-expressing the coenzyme-A ligase-encoding gene (4CL216) from a hybrid poplar and the grapevine resveratrol synthase gene (vst1) in laboratory strains of S. cerevisiae. This yeast has the ability to metabolise p -coumaric acid, a substance already present in grape must. This compound was therefore added to the synthetic media used for the growth of laboratory cultures. Transformants expressing both the 4CL216 and vst1 genes were obtained and tested for production of resveratrol. Following ,-glucosidase treatment of organic extracts for removal of glucose moieties that are typically bound to resveratrol, the results showed that the yeast transformants had produced the resveratrol ,-glucoside, piceid. This is the first report of the reconstruction of a biochemical pathway in a heterologous host to produce resveratrol. [source] Enhancing lignan biosynthesis by over-expressing pinoresinol lariciresinol reductase in transgenic wheatMOLECULAR NUTRITION & FOOD RESEARCH (FORMERLY NAHRUNG/FOOD), Issue 12 2007Allan K. Ayella Abstract Lignans are phenylpropane dimers that are biosynthesized via the phenylpropanoid pathway, in which pinoresinol lariciresinol reductase (PLR) catalyzes the last steps of lignan production. Our previous studies demonstrated that the contents of lignans in various wheat cultivars were significantly associated with anti-tumor activities in APCMin mice. To enhance lignan biosynthesis, this study was conducted to transform wheat cultivars (,Bobwhite', ,Madison', and ,Fielder', respectively) with the Forsythia intermedia PLR gene under the regulatory control of maize ubiquitin promoter. Of 24 putative transgenic wheat lines, we successfully obtained 3 transformants with the inserted ubiquitin-PLR gene as screened by PCR. Southern blot analysis further demonstrated that different copies of the PLR gene up to 5 were carried out in their genomes. Furthermore, a real-time PCR indicated ,17% increase of PLR gene expression over the control in 2 of the 3 positive transformants at T0 generation. The levels of secoisolariciresinol diglucoside, a prominent lignan in wheat as determined by HPLC-MS, were found to be 2.2-times higher in one of the three positive transgenic sub-lines at T2 than that in the wild-type (117.9 ± 4.5 vs. 52.9 ± 19.8 ,g/g, p <0.005). To the best of our knowledge, this is the first study that elevated lignan levels in a transgenic wheat line has been successfully achieved through genetic engineering of over-expressed PLR gene. Although future studies are needed for a stably expression and more efficient transformants, the new wheat line with significantly higher SDG contents obtained from this study may have potential application in providing additive health benefits for cancer prevention. [source] A new catalytic activity from tobacco converting 2-coumaric acid to salicylic aldehydePHYSIOLOGIA PLANTARUM, Issue 3 2007Jacek Malinowski Salicylic acid (SA) mediates plant response to pathogen invasion, resulting in hypersensitive response and in the formation of systemic acquired resistance. It is well known that Nicotiana tabacum and other plants respond to Tobacco Mosaic Virus (TMV) infection by increasing the content of SA but the details of SA biosynthesis are still not fully understood. Generally, SA may originate directly from isochorismate (Arabidopsis thaliana), or its C6,C1 skeleton could be synthesized via the phenylpropanoid pathway by ,-oxidation of trans -cinnamic acid (N. tabacum), 2-coumaric acid (OCA) (Gaulteria procumbens, Lycopersicum esculentum) or by retro-aldol reaction of trans -cinnamoyl-CoA (Hypericum androsaemum). We report here a novel putative enzyme activity from tobacco, salicylic aldehyde synthase (SAS), catalysing non-oxidative formation of salicylic aldehyde (SALD) directly from OCA. This chain-shortening activity is similar to that of 4-hydroxybenzaldehyde synthase from Vanilla planifolia, Lithospermum erythrorhizon, Daucus carota, Solanum tuberosum and Polyporus hispidus but the enzyme differs in the kinetics of the reaction, substrate specificity and requirements for reducing cofactors. SAS activity is constitutively expressed in healthy tobacco leaves and doubles as a result of infection with TMV. Moreover, the product of SAS activity,SALD, applied exogenously on tobacco leaves, stimulates peroxidase activity and enhances resistance to consecutive infection with TMV. These observations could suggest a contribution of SAS and SALD to the response of tobacco to TMV infection. [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] Enzyme activity of the phenylpropanoid pathway as a response to apple scab infectionANNALS OF APPLIED BIOLOGY, Issue 3 2010A. Slatnar The study was performed on apple trees, ,Golden Delicious' cv., which is a scab-susceptible cultivar. The phenolic content of apple fruit was determined in different parts of the peel. The phenolic compounds were analysed in the scab spot, in the tissue around the spot and in the healthy tissue. We determined the concentration of various phenolic compounds and related enzyme activities. Infection with the Venturia inaequalis fungus enhanced the metabolism of phenolic compounds at the scab spot, around the spot and in healthy peel. Compared with the healthy tissue and the tissue around the spot, the scab spot showed higher enzyme activity for all tested enzymes, except for dihydrochalcone 2,- O -glucosyltransferase, which had lower activity in the scab spot. In comparison to the healthy peel, the scab spot showed up to 3.4 times more hydroxycinnamic acids, up to 1.1 times more dihydrochalcones and up to 1.4 times more flavan-3-ols. In contrast, the healthy peel showed up to 1.6 times more flavonols than the scab spot. [source] Identification and expression analysis of a MYB family transcription factor in the parasitic plant Orobanche ramosaANNALS OF APPLIED BIOLOGY, Issue 2 2007C.I. González-Verdejo Abstract MYB proteins are transcription factors (TFs) involved in the regulation of developmental processes in eukaryotes. A number of MYB genes have been identified from plants, but they have not been studied in parasitic plants. In this work, a member of the R2R3 MYB family of TFs was isolated from a complementary DNA library representing different developmental stages of the parasitic plant Orobanche ramosa. The pattern of expression of the gene was studied by in situ hybridisation. Alignment of the deduced Or-MYB1 protein with members of the MYB family showed the highest overall identity with MYB.Ph3 from petunia (Petunia hybrida), NtMYBAS1/S2 from tobacco (Nicotiana tabacum) and AtMYB101 from Arabidopsis thaliana. Amino acid sequence comparisons of DNA-binding domains showed that Or-MYB1 protein forms a closely related group with these proteins. Transcripts of Or-MYB1 were detected during all the developmental stages analysed, and in situ hybridisation showed that the expression was restricted to the parenchymatic cells proximal to the vascular vessels. These findings are consistent with a role of Or-MYB1 during early stages of development of O. ramosa, probably through the phenylpropanoid pathway. [source] Alterations in Taxol Production in Plant Cell Culture via Manipulation of the Phenylalanine Ammonia Lyase PathwayBIOTECHNOLOGY PROGRESS, Issue 6 2002Michelle C. Brincat One approach to increasing secondary metabolite production in plant cell culture is to manipulate metabolic pathways to utilize more resources toward production of one desired compound or class of compounds, such as diverting carbon flux from competing secondary pathways. Since phenylalanine provides both the phenylisoserine side chain and the benzoyl moiety at C-2 of Taxol, we speculated that blockage of the phenylpropanoid pathway might divert phenylalanine into Taxol biosynthesis. We used specific enzyme inhibitors to target the first enzyme in the phenylpropanoid pathway, phenylalanine ammonia lyase (PAL), the critical control point for conversion of l -phenylalanine to trans -cinnamic acid. Cinnamic acid acted quickly in reducing PAL activity by 40,50%, without affecting total protein levels, but it generally inhibited the taxane pathway, reducing Taxol by 90% of control levels. Of the taxanes produced, 13-acetyl-9-dihydro-baccatin III and 9-dihydrobaccatin III doubled as a percentage of total taxanes in C93AD and CO93P cells treated with 0.20 and 0.25 mM cinnamic acid, when all other taxanes were lowered. The PAL inhibitor ,-aminooxyacetic acid (AOA) almost entirely shut down Taxol production at both 0.5 and 1.5 mM, whereas l -,-aminooxy-,-phenylpropionic acid (AOPP) had the opposite effect, slightly enhancing Taxol production at 1 ,M but having no effect at 10 ,M. The discrepancy in the effectiveness of AOA and AOPP and the lack of effect with addition of phenylalanine or benzoic acid derivatives further indicates that the impact of cinnamic acid on Taxol is related not to its effect on PAL but rather to a specific effect on the taxane pathway. On the basis of these results, a less direct route for inhibiting the phenylpropanoid pathway may be required to avoid unwanted side effects and potentially enhance Taxol production. [source] | ||||||||||