DOCSLIB.ORG

The Conversion of Shikimic Acid Into Certain Aromatic Compounds by Cell-Free Extracts of Aerobacter Aerogenes and Escherichia Coli

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Conversion of Shikimic Acid Into Certain Aromatic Compounds by Cell-Free Extracts of Aerobacter Aerogenes and Escherichia Coli Vol. 89 MECHANISM OF ACTION OF CREATINE KINASE 229 Katchalski, E., Fasman, G. D., Simons, E., Blout, E. R., Noda, L., Kuby, S. A. & Lardy, H. A. (1954). J. biol. Gurd, F. R. N. & Koltun, W. L. (1960). Arch. Biophy8. Chem. 209, 203. Biochem. 88, 361. Noltmann, E. A., Mahowald, T. A. & Kuby, S. A. (1962). Koltun, W. H. & Gurd, F. R. N. (1960). J. Amer. chem. J. biol. Chem. 237, 1146. Soc. 81, 301. Poulik, M. D. (1957). Nature, Lond., 180, 1477. Koltun, W. H., Ng, L. & Gurd, F. R. N. (1963). J. biol. Rabin, B. R. & Watts, D. C. (1960). Nature, Lond., 188, Chem. (in the Press). 1163. Krebs, E. G. (1955). In Method8 in Enzymology, vol. 1, Samuels, A. J., Nihei, T. & Noda, L. (1961). Proc. nat. p. 407. Ed. by Colowick, S. P. & Kaplan, N. 0. New Acad. Sci., Wash., 47, 1992. York: Academic Press Inc. Smith, R. M. & Alberty, R. A. (1956). J. Amer. chem. Soc. Kuby, S. A., Noda, L. & Lardy, H. A. (1954a). J. biol. 78, 2376. Chem. 209, 191. Smithies, 0. (1959). Advanc. Protein Chem. 14, 65. Kuby, S. A., Noda, L. & Lardy, H. A. (1954b). J. biol. Watts, D. C. (1961). Ph.D. Thesis: University of London. Chem. 210, 65. Watts, D. C. & Rabin, B. R. (1962). Biochem. J. 85, Mahowald, T. A. & Kuby, S. A. (1960). Fed. Proc. 19, 46. 507. Mahowald, T. A., Noltmann, E. A. & Kuby, S. A. (1962). Watts, D. C., Rabin, B. R. & Crook, E. M. (1961). Biochim. J. biol. Chem. 237, 1535. biophys. Acta, 48, 380. Nihei, T., Noda, L. & Morales, M. F. (1961). J. biol. Chem. Watts, D. C., Rabin, B. R. & Crook, E. M. (1962). Biochem. 236, 3203. J. 82, 412. Biochem. J. (1963) 89, 229 The Conversion of Shikimic Acid into Certain Aromatic Compounds by Cell-Free Extracts of Aerobacter aerogenes and Escherichia coli BY PAULINE N. MORGAN, MARGARET I. GIBSON AND F. GIBSON Bacteriology School, University of Melbourne, Parkville, N. 2, Victoria, Au8tralia (Received 17 December 1962) One of the major unsolved problems in the bio- acid 5-phosphate as substrate, glutamine and NAD, synthesis of aromatic compounds by micro- NADH2 or NADP were required for optimum organisms is the process by which shikimic acid synthesis. 5-phosphate is converted into anthranilic acid. This The present work confirms and extends the results involves the formation of an aromatic ring and the of Srinivasan. The experimental system used was introduction of nitrogen into the molecule and also suitable for examining the biosynthesis of might therefore be expected to take place in several phenylalanine and tyrosine. Part of this work has steps. Although tryptophan auxotrophs that appear been briefly reported (Morgan, Gibson & Gibson, to have a metabolic lesion in this region of the 1962). pathway have been known for many years (Yanof- sky, 1955; Tatum, Bonner & Beadle, 1943; Nyc, MATERIALS AND METHODS Mitchell, Leifer & Langham, 1949; Gots, Koh & 1954), have been no reports of the Organi8ms. Various auxotrophic strains of Escherichia Hunt, there coli and Aerobacter aerogenem were used. These strains were accumulation of possible intermediates by such obtained by penicillin selection (Davis, 1948; Lederberg & cells. Investigations of accumulations by whole Zinder, 1948) after ultraviolet irradiation. The wild-type cells of certain tryptophan auxotrophs yielded strain of A. aerogeneM was first irradiated and a non- mixtures of phenolic compounds (Pittard, Gibson & capsulated variant isolated, this variant being used as the Doy, 1961, 1962; Doy & Gibson, 1961). The relation parent strain for the auxotrophs used. Some details of the of these compounds to tryptophan synthesis, how- strains used are set out in Table 1. ever, was not clear, and it was decided to seek All strains were maintained on meat infusion-agar slopes further information on the conversion of shikimic and subcultured monthly. acid into anthranilic acid by using Prepparation and fradionation of ceU extract. The cells 5-phosphate were grown in the citric acid-mineral salts mixture de- cell-free extracts of bacterial auxotrophs. scribed by Vogel & Bonner (1956), supplemented with Srinivasan (1959) reported briefly on the enzymic 0-16% of glucose, 0.005% of acid-hydrolysed casein and synthesis of anthranilic acid from shikimic acid indole (2 ,ug./ml.). The concentration of indole was made 5-phosphate by cell-free extracts of a mutant strain limiting to encourage de-repression on the tryptophan of E8cherichia coli. He found that, with shikimic pathway. 230 P. N. MORGAN, M. I. GIBSON AND F. GIBSON 1963 Table 1. Strain8 of auxotroph. Substances listed under 'Growth requirement' are the supplements needed for growth in glucose-mineral salts medium. Compound(s) accumulated by Auxotroph Growth requirement cell suspensions Probable site of metabolic lesion A. aerogene. T 17 Tryptophan or indole Anthranilic acid N-(5-Phosphoribosyl)anthranilie acid 1-(o-carboxyphenylamino)- deoxyribulose 5-phosphate* A. aerogenes NC 3 Tryptophan or indole Phenolst Between the last intermediate common or anthranilic acid to the synthesis of all the aromatic amino acids and anthranilic acid E. coli W2-5T Tryptophan or indole Anthranilic acid Anthranilic acid N-(5-phospho- ribosyl)anthranilic acid* * See Doy, Rivera & Srinivasan (1961). t See Pittard, Gibson & Doy (1961). t See Discussion. Quantities (1 1.) of medium were inoculated with about 109 cells from a 6-8 hr. shaken meat-infusion broth culture. Analytical methods The 1 1. cultures in 2 1. flasks were then grown at 370 while Anthranilic acid. In the earlier experiments anthranilic aerated (400 ml./min.) through a central sintered-glass acid was estimated by the Bratton & Marshall (1939) test filter; alternatively, 1 1. quantities were shaken on a rotary for diazotizable amines as described by Gibson & McDougall shaker at 300. Flasks were incubated overnight. No differ- (1961). This method was not specific and it was found more ences were noted in enzyme preparations from the cells convenient to extract anthranilic acid into ethyl acetate grown at different temperatures. and estimate it spectrophotometrically. To 1 ml. of test The cells from each flask were spun down and washed solution was added 0.1 ml. of N-HCI, followed by 4 ml. of once in about 250 ml. of cold 0 9 % NaCl solution. In early ethyl acetate. The tubes were mixed by inversion 10-15 experiments cells were smashed in a Hughes (1951) press at times and centrifuged at a low speed to separate the sol- approximately - 20° and taken up in 10 ml. of 01 M-tris vent. The extinction of the ethyl acetate layer was mea- buffer, pH 7-8. The resulting highly viscous material was sured against an ethyl acetate blank at 336 m,. The con- centrifuged at 24500g for 30 min., the clear supernatant centration of anthranilic acid was calculated by assuming decanted (crude extract) and stored at - 200. In later an extinction coefficient at 336 mu of 4900 (Doy & Gibson, experiments the cells were smashed in a MSE-Mullard 1959). Except for the experiment of Fig. 3, the extraction 500 w ultrasonic disintegrator after resuspending the method was used in the work described. washed cells in 4 ml. ofO lM-tris buffer, pH 7 8/g. wet wt. of Phenylpyruvic acid8. The phenylpyruvic acids were de- cells. After centrifuging as above the supernatants (crude tected after acidification and extraction of reaction mix- extract) were stored at - 200. All the experiments de- tures with ethyl acetate as in the anthranilic acid esti- scribed involving fractionation of enzymes were carried out mations. Immediate spectra and spectra after overnight after ultrasonic treatment. incubation of the ethyl acetate extracts at 30° were re- Nucleic acids were removed from the crude extract by corded. A spectrum with a peak at 288 m,u was obtained the addition of 1-5 ml. of M-MnCl2/20 ml. of extract, stirring after incubation. Extinction at 288 m,u for a given con- for 10 min., centrifuging (24000g for 10 min.) and de- centration of phenylpyruvic acid varied somewhat from canting the supernatant. experiment to experiment although it was consistent in any Ammonium sulphate fractionations were carried out by one experiment. Therefore the amount of phenylpyruvic adding solid (NH4)2S04 to the required saturation (72 g./ acids present in ethyl acetate extracts is expressed in terms 100 ml. was regarded as 100% saturation), stirring for of the extinction at 288 m,u rather than as concentrations. 10 min., centrifuging (24000g for 10 min.), decanting the Adenosine 5'-tripho8phate. Adenosine triphosphate was supernatant and taking up the deposit in 0 1 m-tris buffer, estimated by a micro-method using a luciferin-luciferase pH 7-8, and dialysing against O OlM-tris buffer, pH 7-8, for extract of 'Live-desiccated' Firefly Tails (Sigma Chemical 2 hr. All the operations described above were carried out Co.) and following the method given with the extract. A at 0-50. calibration curve was prepared each time the test was Reagent8. The reagents used with the exception of shiki- performed. The presence of 1 ,tmole of ATP in this test mic acid 5-phosphate, which was kindly given by Dr B. produced approximately 35% transmission. Davis, were obtained commercially and not further purified. Shikimic acid. Two methods were used for the estimation Ribose 5-phosphate solutions were made by treating the of shikimic acid: colorimetric and microbiological. The barium salt with a slight excess of Na2SO4, and removing colorimetric assay was that described by Gaitonde & and washing the precipitate by centrifuging. Boiled crude Gordon (1958). To 3 ml. of solution containing 2-10 ,ug. of extract was made by heating crude extract prepared as shikimic acid was added 0 5 ml. of periodic acid (1%, w/v) above for 5-10 min.
Load more

Recommended publications
  • Metabolomics Reveals the Molecular Mechanisms of Copper Induced
    Article Cite This: Environ. Sci. Technol. 2018, 52, 7092−7100 pubs.acs.org/est Metabolomics Reveals the Molecular Mechanisms of Copper Induced Cucumber Leaf (Cucumis sativus) Senescence † ‡ § ∥ ∥ ∥ Lijuan Zhao, Yuxiong Huang, , Kelly Paglia, Arpana Vaniya, Benjamin Wancewicz, ‡ § and Arturo A. Keller*, , † Key Laboratory of Pollution Control and Resource Reuse, School of Environment, Nanjing University, Nanjing, Jiangsu 210023, China ‡ Bren School of Environmental Science & Management, University of California, Santa Barbara, California 93106-5131, United States § University of California, Center for Environmental Implications of Nanotechnology, Santa Barbara, California 93106, United States ∥ UC Davis Genome Center-Metabolomics, University of California Davis, 451 Health Sciences Drive, Davis, California 95616, United States *S Supporting Information ABSTRACT: Excess copper may disturb plant photosynthesis and induce leaf senescence. The underlying toxicity mechanism is not well understood. Here, 3-week-old cucumber plants were foliar exposed to different copper concentrations (10, 100, and 500 mg/L) for a final dose of 0.21, 2.1, and 10 mg/plant, using CuSO4 as the Cu ion source for 7 days, three times per day. Metabolomics quantified 149 primary and 79 secondary metabolites. A number of intermediates of the tricarboxylic acid (TCA) cycle were significantly down-regulated 1.4−2.4 fold, indicating a perturbed carbohy- drate metabolism. Ascorbate and aldarate metabolism and shikimate- phenylpropanoid biosynthesis (antioxidant and defense related pathways) were perturbed by excess copper. These metabolic responses occur even at the lowest copper dose considered although no phenotype changes were observed at this dose. High copper dose resulted in a 2-fold increase in phytol, a degradation product of chlorophyll.
    [Show full text]
  • Class 11 Biology Chapter- 13 Respiration in Plants
    CLASS 11 BIOLOGY CHAPTER- 13 RESPIRATION IN PLANTS CELLULAR RESPIRATION: The process of conversion of the chemical energy of organic substances into a metabolically usable energy within living cells is called cellular respiration. TYPES OF CELLULAR RESPIRATION: (i) Aerobic respiration: The process of respiration which requires molecular oxygen. (ii) Anaerobic respiration: The process of respiration which does not require molecular oxygen and occurs in the cytoplasm. MECHANISM OF RESPIRATION: Following are the steps- 1. GLYCOLYSIS / EMP Pathway: It involves a series of closely integrated reactions in which hexose sugars(usually glucose) are converted into pyruvic acid. It is common in both aerobic and anaerobic reactions. It occurs in the cytoplasm. It does not require oxygen. Gollowing are the steps of GLYCLOLYSIS: (i) Conversion of glucose to Fructose-1,6-diphosphate: First phosphorylation: Glucose is converted to Glucose -6-phosphate in the presence of enzyme hexokinase and Mg++ ions and energy in the form of ATP. Isomerization: Glucose-6-phosphate is converted to Fructose-6-phosphate in the presence of phosphohexoisomerase. Second phosphorylation: Fructose-6-phosphate is converted to Fructose-1,6- diphosphate by the use of energy in the form of ATP. (ii) Formation of pyruvic acid from fructose -1,6-diphosphate: Cleavage: Fructose-1,6-diphosphate splits into 3-phosphoglyceraldehyde and Dihydroxyacetone phosphate in the presence of enzyme aldolase. Phosphorylation and oxidative dehydrogenase: 3-phosphoglyceraldehyde is converted to 1,3-biphosphoglyceric acid. ATP generation(first): 1,3-biphosphoglyceric acid is converted to 3-phosphoglyceric acid in the presence of Mg++ and phosphoglycerokinase . Isomerization: 3-phosphoglyceric acid is converted to 2-phosphoglyceric acid in the presence of Mg++.
    [Show full text]
  • I (Theoretical Organic Chemistry-I)
    M.Sc. Organic Chemistry Semester – III Course Code: PSCHO301 Paper - I (Theoretical organic chemistry-I) Unit 1 Organic reaction mechanisms [15L] 1.1 Organic reactive intermediates, methods of generation, structure, stability [5L] and important reactions involving carbocations, nitrenes, carbenes, arynes and ketenes. 1.2 Neighbouring group participation: Mechanism and effects of anchimeric [3L] assistance, NGP by unshared/ lone pair electrons, π-electrons, aromatic rings, σ-bonds with special reference to norbornyl and bicyclo[2.2.2]octyl cation systems (formation of non-classical carbocation) 1.3 Role of FMOs in organic reactivity: Reactions involving hard and soft [2L] electrophiles and nucleophiles, ambident nucleophiles, ambident electrophiles, the α effect. 1.4 Pericyclic reactions: Classification of pericyclic reactions; thermal and [5L] photochemical reactions. Three approaches: Evidence for the concertedness of bond making and breaking Symmetry-Allowed and Symmetry-Forbidden Reactions – The Woodward-Hoffmann Rules-Class by Class The generalised Woodward-Hoffmann Rule Explanations for Woodward-Hoffmann Rules The Aromatic Transition structures [Huckel and Mobius] Frontier Orbitals Correlation Diagrams, FMO and PMO approach Molecular orbital symmetry, Frontier orbital of ethylene, 1,3 butadiene, 1,3,5 hexatriene and allyl system. Unit 2 Pericyclic reactions [15L] 2.1 Cycloaddition reactions: Supra and antra facial additions, 4n and 4n+2 [7L] systems, 2+2 additions of ketenes. Diels-Alder reactions, 1, 3-Dipolar cycloaddition and cheletropic reactions, ene reaction, retro-Diels-Alder reaction, regioselectivity, periselectivity, torquoselectivity, site selectivity and effect of substituents in Diels-Alder reactions. Other Cycloaddition Reactions- [4+6] Cycloadditions, Ketene Cycloaddition, Allene Cycloadditions, Carbene Cycloaddition, Epoxidation and Related Cycloadditions. Other Pericyclic reactions: Sigmatropic Rearrangements, Electrocyclic Reactions, Alder ‘Ene’ Reactions.
    [Show full text]
  • Effects of A-Aminooxy-ß-Phenylpropionic Acid on Phenylalanine Metabolism in /R-Fluorophenylalanine Sensitive and Resistant Toba
    Effects of a-Aminooxy-ß-phenylpropionic Acid on Phenylalanine Metabolism in /r-Fluorophenylalanine Sensitive and Resistant Tobacco Cells Jochen Berhn and Brigitte Vollmer Lehrstuhl fur Biochemie der Pflanzen, Westfälische Wilhelms-Universität Münster, West Germany Z. Naturforsch. 34 c, 770 — 775 (1979); received May 18, 1979 Nicotiana tabacum, Suspension Cultures, a-Aminooxy-ß-phenylpropionic Acid, Chorismate Mu- tase, Phenylalanine, Pool Sizes A ^-fluorophenylalanine (PFP) resistant cell line with high phenylalanine ammonia lyase (PAL) activity and wild type cells with low PAL activity were compared in their responses to PAL inhibition by a-aminooxy-/?-phenylpropionic acid (AOP). Inhibition of PAL reduced the levels of the main phenolic compounds to 30% of the controls. Free phenylalanine pools increased 17 fold in the resistant line and 6 fold in the sensitive line, respectively. The accumulation of phenylalani­ ne did not reduce the flow of labeled shikimic acid into the aromatic amino acids tyrosine and phenylalanine. The results are discussed with respect to the feedback inhibition of chorismate mu- tase activity by phenylalanine and tyrosine in both cell lines. Amino acid analog resistant cell lines normally Therefore the cells were grown in the presence of a have an increased pool size of the corresponding na­ specific inhibitor of PAL, a-aminooxy-yß-phenyl- tural amino acid due to a lessened feedback control propionic acid (AOP) [ 6 ], Inhibition of PAL should in the biosynthetic pathway [1]. A tobacco cell line result in distinct changes of phenylalanine pools in resistant to / 7-fluorophenylalanine (PFP), however, sensitive and resistant cells and/or should reduce the did not fit into this frame [2-5], The main reason for flow of carbon through the shikimate pathway.
    [Show full text]
  • Anaerobic Performance and Metabolism of the Hyperthyroid Heart
    Anaerobic Performance and Metabolism of the Hyperthyroid Heart Rum A. ALTSCHULD, ALAN WEISS, FRED A. KRUGER, and ARNOLD M. WEISSLER From the Department of Medicine, Ohio State University College of Medicine, Columbus, Ohio 43210 A B S T R A C T Anaerobically perfused hearts from rats all consequences of hyperthyroilism (7-9). These with experimentally induced hyperthyroidism exhibited changes are accompanied by increased cardiac oxygen accelerated deterioration of pacemaker activity and ven- consumption and free fatty acid utilization, whereas tricular performance. The diminished anaerobic per- glucose uptake and oxidation are decreased (10, 11). formance of hyperthyroid hearts was associated with In view of the recent demonstration in our laboratory decreased adenosine triphosphate (ATP) levels and a of the importance of glucose metabolism in preserving reduced rate of anaerobic glycolysis as reflected in de- structure and function of the anoxic heart (12) and the creased lactic acid production during 30 min of anoxic reported inhibition of glucose metabolism in hyperthy- perfusion. roid hearts (6), it seemed desirable to determine whether Studies on whole heart homogenates demonstrated in- anaerobic metabolism is altered in the intact hearts hibition at the phosphofructokinase (PFK) step of the from hyperthyroid animals and to ascertain what effects glycolytic pathway. Such inhibition was not demonstrated these metabolic alterations have on anoxic performance in the hyperthyroid heart cytosol. It is postulated that of the heart. an inhibitor of PFK which resides dominantly in the particulate fraction is probably responsible for the di- METHODS minished anaerobic glvcolvsis and performance of the Male Wistar rats fed ad lib. with Purina laboratory chow hyperthyroid heart.
    [Show full text]
  • Download Product Insert (PDF)
    PRODUCT INFORMATION Phosphoenolpyruvic Acid (potassium salt) Item No. 19192 CAS Registry No.: 4265-07-0 Formal Name: 2-(phosphonooxy)-2-propenoic acid, O HO monopotassium salt Synonym: PEP-K P OH - MF: C H O P • K O O 3 4 6 + FW: 206.1 • K O Purity: ≥95% Supplied as: A crystalline solid Storage: -20°C Stability: As supplied, 2 years from the QC date provided on the Certificate of Analysis, when stored properly Laboratory Procedures Phosphoenolpyruvic acid (potassium salt) is supplied as a crystalline solid. Phosphoenolpyruvic acid (potassium salt) is sparingly soluble in organic solvents such as ethanol, DMSO, and dimethyl formamide. For biological experiments, we suggest that organic solvent-free aqueous solutions of phosphoenolpyruvic acid (potassium salt) be prepared by directly dissolving the crystalline solid in aqueous buffers. The solubility of phosphoenolpyruvic acid (potassium salt) in PBS, pH 7.2, is approximately 10 mg/ml. We do not recommend storing the aqueous solution for more than one day. Description Phosphoenolpyruvic acid plays a role in both glycolysis and gluconeogenesis. During glycolysis, it is formed by the action of enolase on 2-phosphoglycerate and is metabolized to pyruvate by pyruvate kinase.1 One molecule of ATP is formed during its metabolism in this pathway. During gluconeogenesis, it is formed from phosphoenolpyruvate carboxykinase-catalyzed oxaloacetate decarboxylation and GTP hydrolysis.1-3 In plants, it is metabolized to form aromatic amino acids and also serves as a substrate for phosphoenolpyruvate carboxylase-catalyzed carbon fixation.4 References 1. Muñoz, M.E. and Ponce, E. Pyruvate kinase: Current status of regulatory and functional properties.
    [Show full text]
  • The Peculiar Glycolytic Pathway in Hyperthermophylic Archaea: Understanding Its Whims by Experimentation in Silico
    Article The Peculiar Glycolytic Pathway in Hyperthermophylic Archaea: Understanding Its Whims by Experimentation In Silico Yanfei Zhang 1, Theresa Kouril 2,3, Jacky L. Snoep 3,4,5, Bettina Siebers 2, Matteo Barberis 1 and Hans V. Westerhoff 1,4,5,* 1 Synthetic Systems Biology and Nuclear Organization, Swammerdam Institute for Life Sciences, University of Amsterdam, 1098 XH Amsterdam, The Netherlands; [email protected] (Y.Z.); [email protected] (M.B.) 2 Molecular Enzyme Technology and Biochemistry (MEB), Biofilm Centre, Centre for Water and Environment Research (CWE), University Duisburg—Essen, Universitätsstr. 5, 45141 Essen, Germany; [email protected] (T.K.); [email protected] (B.S.) 3 Department of Biochemistry, University of Stellenbosch, Stellenbosch 7602, South Africa; [email protected] 4 The Manchester Centre for Integrative Systems Biology, Manchester Institute for Biotechnology, School for Chemical Engineering and Analytical Science, University of Manchester, Manchester M1 7DN, UK 5 Department of Molecular Cell Physiology, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands * Correspondence: [email protected]; Tel.: +31-20-598-7230 Academic Editor: Samuel De Visser Received: 12 March 2017; Accepted: 13 April 2017; Published: 20 April 2017 Abstract: Mathematical models are key to systems biology where they typically describe the topology and dynamics of biological networks, listing biochemical entities and their relationships with one another. Some (hyper)thermophilic Archaea contain an enzyme, called non- phosphorylating glyceraldehyde-3-phosphate dehydrogenase (GAPN), which catalyzes the direct oxidation of glyceraldehyde-3-phosphate to 3-phosphoglycerate omitting adenosine 5′-triphosphate (ATP) formation by substrate-level-phosphorylation via phosphoglycerate kinase.
    [Show full text]
  • 102 4. Biosynthesis of Natural Products Derived from Shikimic Acid
    102 4. Biosynthesis of Natural Products Derived from Shikimic Acid 4.1. Phenyl-Propanoid Natural Products (C6-C3) The biosynthesis of the aromatic amino acids occurs through the shikimic acid pathway, which is found in plants and microorganisms (but not in animals). We (humans) require these amino acids in our diet, since we are unable to produce them. For this reason, molecules that can inhibit enzymes on the shikimate pathway are potentially useful as antibiotics or herbicides, since they should not be toxic for humans. COO COO NH R = H Phenylalanine 3 R = OH Tyrosine R NH3 N Tryptophan H The aromatic amino acids also serve as starting materials for the biosynthesis of many interesting natural products. Here we will focus on the so-called phenyl-propanoide (C6-C3) natural products, e.g.: OH OH OH HO O HO OH HO O Chalcone OH O a Flavone OH O OH O a Flavonone OH OH Ar RO O O O HO O O OH O OR OH Anthocyanine OH O a Flavonol Podophyllotoxin MeO OMe OMe OH COOH Cinnamyl alcohol HO O O Cinnamic acid OH (Zimtsäure) Umbellierfone OH a Coumarin) MeO OH O COOH HO Polymerization OH Wood OH HO OH O OH MeO OMe Shikimic acid O HO 4.2. Shikimic acid biosynthesis The shikimic acid pathway starts in carbohydrate metabolism. Given the great social and industrial significance of this pathway, the enzymes have been intensively investigated. Here we will focus on the mechanisms of action of several key enzymes in the pathway. The following Scheme shows the pathway to shikimic acid: 103 COO- COO- Phosphoenolpyruvate HO COO- 2- O O3P-O 2- O3P-O DHQ-Synthase
    [Show full text]
  • Development of an Assay for a Diels-Alderase Enzyme
    Development of an Assay for a Diels-Alderase Enzyme A Thesis submitted in part fulfilment of the requirements of the degree of Doctor of Philosophy Graeme Douglas McAllister Department of Chemistry University of Glasgow Glasgow G12 8QQ February 2000 ©Graeme D. McAllister ProQuest Number: 13818648 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a com plete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion. uest ProQuest 13818648 Published by ProQuest LLC(2018). Copyright of the Dissertation is held by the Author. All rights reserved. This work is protected against unauthorized copying under Title 17, United States C ode Microform Edition © ProQuest LLC. ProQuest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, Ml 48106- 1346 Dedicated to iny family "Do they give Nobel prizes for attempted chemistry? Do they!?" from 'The Simpsons' by Matt Groening Acknowledgements First of all my sincerest thanks go to my supervisor, Dr Richard Hartley, for his expert guidance over the last 3 years. I would also like to thank Dr Mike Dawson and Dr Andy Knaggs of GlaxoWellcome for their supervision and ideas in the biological areas of this project, and for helping a chemist adjust to life in a biology lab! Dr Chris Brett of the University of Glasgow and Mrs Jyoti Vithlani of GlaxoWellcome deserve a mention for all their expertise in the growing of cell cultures and for helping me in the feeding studies.
    [Show full text]
  • 8.2 Shikimic Acid Pathway
    CHAPTER 8 © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FORAromatic SALE OR DISTRIBUTION and NOT FOR SALE OR DISTRIBUTION Phenolic Compounds © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION CHAPTER OUTLINE Overview Synthesis and Properties of Polyketides 8.1 8.5 Synthesis of Chalcones © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC 8.2 Shikimic Acid Pathway Synthesis of Flavanones and Derivatives NOT FOR SALE ORPhenylalanine DISTRIBUTION and Tyrosine Synthesis NOT FOR SALESynthesis OR DISTRIBUTION and Properties of Flavones Tryptophan Synthesis Synthesis and Properties of Anthocyanidins Synthesis and Properties of Isofl avonoids Phenylpropanoid Pathway 8.3 Examples of Other Plant Polyketide Synthases Synthesis of Trans-Cinnamic Acid Synthesis and Activity of Coumarins Lignin Synthesis Polymerization© Jonesof Monolignols & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC Genetic EngineeringNOT FOR of Lignin SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION Natural Products Derived from the 8.4 Phenylpropanoid Pathway Natural Products from Monolignols © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION 119 © Jones & Bartlett Learning, LLC.
    [Show full text]
  • Phosphoenolpyruvic Acid, an Intermediate of Glycolysis
    Journal of Health Science, 56(6) 727–732 (2010) 727 — Research Letter — Phosphoenolpyruvic Acid, drate with anti-oxidant properties and suggest that PEP is potentially useful as an organ preservation an Intermediate of Glycolysis, agent in clinical transplantation. Attenuates Cellular Injury Induced by Hydrogen Peroxide Key words —— Phosphoenolpyruvate, carbohydrate, oxidative stress, transplantation, cellular injury and 2-Deoxy-D-glucose in the Porcine Proximal Kidney Tubular INTRODUCTION Cell Line, LLC-PK1 Ischemia/reperfusion injury, which results from a ∗, a Yuki Kondo, Yoichi Ishitsuka, the interruption of blood flow and subsequent organ b, c a Daisuke Kadowaki, Minako Nagatome, reperfusion, contributes to graft dysfunction and a a Yusuke Saisho, Masataka Kuroda, acute graft rejection; it is, therefore, a major prob- c a Sumio Hirata, Mitsuru Irikura, lem in clinical organ transplantation.1–3) Oxidative d a, c Naotaka Hamasaki, and Tetsumi Irie stress and compromised energy metabolism are as- sociated, at least in part, with ischemia/reperfusion a Department of Clinical Chemistry and Informatics, Gradu- injury.4–7) To inhibit this, organ preservation so- ate School of Pharmaceutical Sciences, Kumamoto University, lutions such as Euro-Collins solution and Univer- 5–1 Oe-honmachi, Kumamoto 862–0973, Japan, bDepartment sity of Wisconsin solution are used.8–10) These so- of Biopharmaceutics, Graduate School of Pharmaceutical Sci- ences, Kumamoto University, 5–1 Oe-honmachi, Kumamoto lutions are generally composed of electrolytes and 862–0973,
    [Show full text]
  • Biol. Pharm. Bull. 42(1): 123-129 (2019)
    Vol. 42, No. 1 Biol. Pharm. Bull. 42, 123–129 (2019) 123 Regular Article Targeting Pyruvate Kinase M2 and Hexokinase II, Pachymic Acid Impairs Glucose Metabolism and Induces Mitochondrial Apoptosis Guopeng Miao,* Juan Han, Jifeng Zhang, Yihai Wu, and Guanhe Tong* Department of Bioengineering, Huainan Normal University; Huainan 232038, China. Received September 20, 2018; accepted October 18, 2018; advance publication released online October 30, 2018 Pachymic acid (PA), a triterpenoid from Poria cocos, has various pharmacological effects, including anti-inflammatory, anti-cancer, anti-aging, and insulin-like properties. PA has gained considerable research attention, but the mechanism of its anti-cancer effects remains unclear. In this study, pyruvate kinase M2 (PKM2) was discovered as a PA target via the drug affinity responsive target stability. Molecular docking and enzyme assay revealed that PA is a competing activator of PKM2, and mimics the natural activator, fructose-1,6-bisphosphate. PKM2 activation should augment the flux of glycolysis. However, decreased glu- cose uptake and lactate production after PA treatment was observed in SK-BR-3 breast carcinoma cells, indicating a blockage or downregulation of glycolysis. The potential of previously reported triterpenoids in blocking hexokinase II (HK2) activity inspired us to investigate the inhibition effect of PA on HK2 activity. Molecular docking and enzyme assay confirmed that PA was an inhibitor of HK2, with an IC50 of 5.01 µM. The possible consequences of glycometabolic regulation by PA, such as dissociation of HK2 from the mito- chondria, release of mitochondrial cytochrome (Cyt) c, depletion of ATP, and generation of reactive oxygen species, were further validated.
    [Show full text]