DOCSLIB.ORG

Functional Genomics of the Bacterial Degradation of the Emerging Water Contaminants: 1,4-Dioxane and N-Nitrosodimethylamine (NDMA)

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Functional Genomics of the Bacterial Degradation of the Emerging Water Contaminants: 1,4-Dioxane and N-Nitrosodimethylamine (NDMA) Functional genomics of the bacterial degradation of the emerging water contaminants: 1,4-dioxane and N-nitrosodimethylamine (NDMA) by Christopher Michael Sales A dissertation submitted in partial satisfaction of the requirements for the degree of Doctor of Philosophy in Engineering-Civil and Environmental Engineering in the Graduate Division of the University of California, Berkeley Committee in charge: Professor Lisa Alvarez-Cohen, Chair Professor Kara L. Nelson Professor Mary K. Firestone Spring 2012 Functional genomics of the bacterial degradation of the emerging water contaminants: 1,4-dioxane and N-nitrosodimethylamine (NDMA) Copyright 2012 By Christopher Michael Sales Abstract Functional genomics of the bacterial degradation of the emerging water contaminants: 1,4-dioxane and N-nitrosodimethylamine (NDMA) by Christopher Michael Sales Doctor of Philosophy in Engineering - Civil & Environmental Engineering University of California, Berkeley Professor Lisa Alvarez-Cohen, Chair The emerging water contaminants 1,4-dioxane and N-nitrosodimethylamine (NDMA) are toxic and classified as probable human carcinogens. Both compounds are persistent in the environment and are highly mobile in groundwater plumes due to their hydrophilic nature. The major source of 1,4-dioxane is due to its use as a stabilizer in the chlorinated solvent 1,1,1-trichloroethane. The presence of NDMA as a water contaminant is related to the release of rocket fuels and its formation in the disinfection of water and wastewater. Prior studies have demonstrated that bacteria expressing monooxygenases are capable of degrading 1,4-dioxane and NDMA. While growth on 1,4-dioxane as a sole carbon and energy source has been reported in Pseudonocardia dioxanivorans CB1190 and Pseudonocardia benzenivorans B5, it is also co-metabolically degradable by a variety of monooxygenase-expressing strains. In contrast, NDMA has only been observed to biode- grade co-metabolically after growth on some monooxygenase-inducing substrates. The fastest rates of NDMA degradation occur in Rhodococcus sp. RR1 and Mycobacterium vaccae JOB5 after growth on propane. Pathways have been proposed for 1,4-dioxane biodegradation in P. dioxanivorans and for NDMA biodegradation in propane-induced Rhodococcus sp. RR1 based only on identified intermediates. The overall goal of this dissertation is to gain a better understanding of the genes and biological pathways respon- sible for degradation of 1,4-dioxane and NDMA. Due to the lack of molecular sequence information for organisms capable of growth on 1,4-dioxane, the genome of P. dioxanivorans strain CB1190 was sequenced in this study. The genome has a total size of 7,440,794 bp and consists of four replicons: a circular chromosome, a circular plasmid pPSED01, an unclosed circular plasmid pPSED02, and a linear plasmid pPSED03. Analysis of this genome sequence revealed the presence of eight multicomponent monooxygenases: a propane monooxygenase, a phenol mono- oxygenase, a 4-hydroxyphenylacetate monooxygenase, four aromatic (toluene) mono- oxygenases, and a tetrahydrofuran (THF) monooxygenase. A total of 92 genes identified as putative dioxygenases were identified. Protein-encoding genes involved in transport systems, signal transduction systems, secretion systems, and heavy-metal and antibiotic resistance were identified. The presence of pathways for carbon and nitrogen metabolism was examined. A complete Calvin-Benson-Bassham carbon fixation pathway was found and a number of carboxylases that function in other carbon fixation pathways were identi- fied. AlthoughP. dioxanivorans has been reported to fix nitrogen, no nitrogenase genes 1 were found. The genome sequence of P. dioxanivorans was compared to other sequenced genomes of members in the family Pseudonocardiaceae, including Amycolatopsis medi- terranei S699, Amycolatopsis mediterranei U32, Pseudonocardia sp. P1, Pseudonocardia sp. P2, Saccharomonaspora azurea NA-128, Saccharomonospora paurometabolica YIM 9007, Sacharomonospora viridis DSM43017, Saccharopolyspora erythraea NRRL2338, and Saccharopolyspora spinosa NRRL18395. Genome-aided approaches were employed to identify the protein-encoding genes in- volved in the metabolic degradation of 1,4-dioxane in P. dioxanivorans. These ap- proaches included whole genome expression microarray transcriptomics, quantitative reverse transcriptase PCR (qRT-PCR), enzyme assays, and heterologous expression clones. Genes differentially expressed during growth on 1,4-dioxane, glycolate (a pre- viously identified degradation intermediate), and pyruvate (control) were analyzed to determine genes differentially expressed and involved in the metabolism of 1,4-dioxane. Based on the differentially expressed genes, the 1,4-dioxane degradation pathway was revised and annotated with the enzymes known to catalyze specific transformation steps. Up-regulation of genes were confirmed and quantified by qRT-PCR. Transcriptional analyses, isotopic tracer analyses with 1,4-[U-13C]-dioxane, and glyoxylate carboligase enzymatic activity assays confirmed the role of glyoxylate as a central intermediate in the degradation of 1,4-dioxane. Specifically, transcriptional analyses indicated that the THF monooxygenase, encoded by thmADBC, is up-regulated during growth on 1,4-dioxane and THF. Furthermore, Rhodococcus jostii RHA1 clones heterologously expressing the P. dioxanivorans genes thmADBC demonstrated 1,4-dioxane and THF degradation activity. A surprising result with the THF monooxygenase expressing clones was the accumulation of the intermediate β-hydroxyethoxyacetic acid (HEAA). This result, combined with the non-inhibitory effect of acetylene on HEAA degradation by 1,4-dioxane grown P. dioxa- nivorans indicates that a monooxygenase is not involved in the transformation of HEAA as previously hypothesized. Transcriptomic microarray analysis of THF- and succinate- grown cells led to the first proposed THF metabolic degradation pathway forP. dioxaniv- orans. A novel finding of this transcriptomic microarray analysis was the identification of an alcohol dehydrogenase up-regulated during growth on THF that could catalyze the conversion of 2-hydroxytetrahydrofuran to γ-butyrolactone. The genome sequence of Rhodococcus jostii RHA1 was utilized to determine the pro- pane-induced monooxygenase responsible for NDMA degradation. The degradation of NDMA was characterized in R. jostii RHA1 grown on propane and on non-inducing substrates (LB medium, soy broth, and pyruvate). Propane enhanced the removal rate of NDMA by nearly two orders of magnitude compared to constitutive degradation during growth on non-inducing substrate. Transcriptomic microarray and qRT-PCR analyses demonstrated that propane elicits the up-regulation of a propane monooxygenase gene cluster. Genetic knockouts of the prmA gene encoding the large hydroxylase component of the propane monooxygenase were unable to grow on propane and degrade NDMA. These results indicate that the propane monooxygenase is responsible for NDMA degra- dation by R. jostii and explain the enhanced co-metabolic degradation of NDMA in the presence of propane. With the newly gained knowledge of the role of propane monooxy- genase in NDMA degradation, oligonucleotide degenerate primers targeting prmA were designed and were identify and quantify the presence of propane monooxygenase genes 2 in Rhodoccocus sp. RR1 and M. vaccae JOB5. A homolog to prmA was found in Rhodoc- cocus sp. RR1 but not in M. vaccae JOB5. The prmA gene in Rhodococcus sp. RR1 was up-regulated during growth on propane relative to pyruvate. Characterization of the kinet- ics of propane-enhanced NDMA degradation showed that Rhodococcus sp. RR1 and M. vaccae possess similar maximum transformation rates (44 ± 5 and 28 ± 3 mg NDMA(mg -1 -1 protein) h , respectively). However, a comparison of half saturation constants (Ks,n) and NDMA degradation in the presence of propane revealed pronounced differences be- -1 tween the strains. The Ks,n for Rhodococcus sp. RR1 was 36 ± 10 mg NDMA L while the propane concentration needed to inhibit NDMA rates by 50% (Kinh) occurred at 7,700 mg propane L-1 (R2 = 0.9669). In contrast, M. vaccae had a markedly lower affinity for -1 NDMA verses propane with a calculated Ks,n of 2,200 ± 1,000 mg NDMA L and Kinh of 120 mg propane L-1 (R2 = 0.9895). Differences between the propane monooxygenases in Rhodococcus sp. RR1 and the unidentified enzyme(s) inM. vaccae may explain the disparities in NDMA degradation and inhibition kinetics between these strains. 3 To Hilary i ii Table of Contents Abstract………………………………………………………………………………….....1 Dedication………………………………………………………………………………….i Table of Contents…………………………………………………………………………iii List of Figures…………………………………………………………………………...viii List of Table………………………………………………………………………...…..…x Acknowledgements…………………………………………………………………...….xii Chapter 1: Introduction and Objectives...........................................................................1 1.1 Introduction……………………………………………………………………………2 1.2 Research goal and specific objecties.......……………………………………………...3 1.3 Dissertation overview……………………………………………………………........4 Chapter 2: Background and Literature Review..............................................................5 2.1 Introduction…………………………………………………………………...............6 2.2 1,4-dioxane and NDMA as emerging contaminants…………………………..............7 2.2.1 Drinking water regulations and health effects of 1,4-dioxane…………................7 2.2.2 Uses of 1,4-dioxane………………………………………………….....................9 2.2.3 Occurrence
Load more

Recommended publications
  • METACYC ID Description A0AR23 GO:0004842 (Ubiquitin-Protein Ligase
    Electronic Supplementary Material (ESI) for Integrative Biology This journal is © The Royal Society of Chemistry 2012 Heat Stress Responsive Zostera marina Genes, Southern Population (α=0.
    [Show full text]
  • The Pennsylvania State University the Graduate School Department
    The Pennsylvania State University The Graduate School Department of Chemistry SUBSTRATE POSITIONING AND CHANNELING OF ESCHERICHIA COLI QUINOLINATE SYNTHASE A Thesis in Chemistry by Lauren A. Sites 2012 Lauren A. Sites Submitted in Partial Fulfillment of the Requirements for the Degree of Master of Science December 2012 ii The thesis of Lauren A. Sites was reviewed and approved* by the following: Squire J. Booker Associate Professor of Chemistry Thesis Advisor Carsten Krebs Professor of Chemistry Scott A. Showalter Assistant Professor of Chemistry Kenneth S. Feldman Professor of Chemistry Head of Department *Signatures are on file in the Graduate School iii ABSTRACT The essential cofactor nicotinamide adenine dinucleotide (NAD) is consumed in many metabolic reactions in the cell, necessitating the need to synthesize NAD. In most bacteria, the de novo pathway to form NAD begins with two unique enzymes that have been extensively studied herein. The first enzyme in the pathway, L-aspartate oxidase, performs a two-electron oxidation of L-aspartate to form iminoaspartate. This flavin containing enzyme can undergo multiple catalytic turnovers given the oxidants, fumarate or molecular oxygen, to afford the oxidized form of the enzyme. The second enzyme in the pathway, quinolinate synthase or NadA, condenses iminoaspartate and dihydroxyacetone phosphate to form quinolinic acid, the backbone of the pyridine ring of NAD. Many have postulated that these two enzymes can operate as an enzyme complex, yet no substantial evidence of this complex has been demonstrated. Investigations to examine the possible protein-protein interactions of the two enzymes were carried out, yet no obvious interaction was seen by the techniques employed.
    [Show full text]
  • The Safety Evaluation of Food Flavouring Substances
    Toxicology Research View Article Online REVIEW View Journal The safety evaluation of food flavouring substances: the role of metabolic studies Cite this: DOI: 10.1039/c7tx00254h Robert L. Smith,a Samuel M. Cohen, b Shoji Fukushima,c Nigel J. Gooderham,d Stephen S. Hecht,e F. Peter Guengerich, f Ivonne M. C. M. Rietjens,g Maria Bastaki,h Christie L. Harman,h Margaret M. McGowenh and Sean V. Taylor *h The safety assessment of a flavour substance examines several factors, including metabolic and physio- logical disposition data. The present article provides an overview of the metabolism and disposition of flavour substances by identifying general applicable principles of metabolism to illustrate how information on metabolic fate is taken into account in their safety evaluation. The metabolism of the majority of flavour substances involves a series both of enzymatic and non-enzymatic biotransformation that often results in products that are more hydrophilic and more readily excretable than their precursors. Flavours can undergo metabolic reactions, such as oxidation, reduction, or hydrolysis that alter a functional group relative to the parent compound. The altered functional group may serve as a reaction site for a sub- sequent metabolic transformation. Metabolic intermediates undergo conjugation with an endogenous agent such as glucuronic acid, sulphate, glutathione, amino acids, or acetate. Such conjugates are typi- Received 25th September 2017, cally readily excreted through the kidneys and liver. This paper summarizes the types of metabolic reac- Accepted 21st March 2018 tions that have been documented for flavour substances that are added to the human food chain, the DOI: 10.1039/c7tx00254h methodologies available for metabolic studies, and the factors that affect the metabolic fate of a flavour rsc.li/toxicology-research substance.
    [Show full text]
  • Pseudonocardia Acaciae Sp. Nov., Isolated from Roots of Acacia Auriculiformis A
    Pseudonocardia acaciae sp. nov., isolated from roots of Acacia auriculiformis A. Cunn. ex Benth. Page 1 of 2 Pseudonocardia acaciae sp. nov., isolated from roots of Acacia auriculiformis A. Cunn. ex Benth. 123 Kannika Duangmal , Arinthip Thamchaipenet , Atsuko Matsumoto and 3 Yoko Takahashi - Author Affiliations 1Department of Microbiology, Faculty of Science, Kasetsart University, Chatuchak, Bangkok 10900, Thailand 2Department of Genetics, Faculty of Science, Kasetsart University, Chatuchak, Bangkok 10900, Thailand 3Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8641, Japan Correspondence Kannika Duangmal [email protected] or [email protected] Abstract A novel Gram-positive-staining actinomycete designated strain GMKU095T was isolated from surface-sterilized roots of Acacia auriculiformis A. Cunn. ex Benth. (earpod wattle). The organism produced branching mycelium. The spores were non-motile and had a spiny surface. Growth of strain GMKU095T occurred at 18– 42 °C, pH 5.0–8.0 and at NaCl concentrations up to 5 %. Whole-cell hydrolysates contained arabinose and galactose as major characteristic sugars. The diagnostic diamino acid of the peptidoglycan was meso-diaminopimelic acid. The glycan moiety of the murein contained acetyl residues. The menaquinone was MK-8(H4); mycolic acids were not detected. The G+C content of the DNA was 71.6 mol%. iso- C16 : 0 was detected as the major cellular fatty acid. Comparative studies of 16S rRNA gene sequences indicated that the strain was phylogenetically related to members of the genus Pseudonocardia. The most closely related type strain is Pseudonocardia spinosispora IMSNU 50581T , which is 96.2 % similar in 16S rRNA gene sequence.
    [Show full text]
  • Phosphatidylinositol-3-Kinase in Tomato (Solanum Lycopersicum. L) Fruit and Its Role in Ethylene Signal Transduction and Senescence
    Phosphatidylinositol-3-Kinase in Tomato (Solanum lycopersicum. L) Fruit and Its Role in Ethylene Signal Transduction and Senescence by Mohd Sabri Pak Dek A Thesis presented to The University of Guelph In partial fulfilment of requirements for the degree of Doctor of Philosophy in Plant Agriculture Guelph, Ontario, Canada © Mohd Sabri Pak Dek,June, 2015 ABSTRACT PHOSPHATIDYLINOSITOL-3-KINASE IN TOMATO (SOLANUM LYCOPERSICUM. L) FRUIT AND ITS ROLE IN ETHYLENE SIGNAL TRANSDUCTION AND SENESCENCE Mohd Sabri Pak Dek Co-Advisors: University of Guelph, 2015 Professor G. Paliyath Professor J. Subramanian The ripening process is initiated by ethylene through a signal transduction cascade leads to the expression of ripening-related genes and catabolism of membrane, cell wall, and storage components. One of the minor components in membrane phospholipids is phosphatidylinositol (PI). Phosphatidylinositol-3-kinase (PIK) is an enzyme that phosphorylates PI at the 3-OH position of inositol head group to produce phosphatidylinositol 3-phosphate (PI3P). Phosphorylation of PI may be an early event in the ethylene signal transduction pathway that generates negatively charged domains on the plasma membrane. PI3P domains may potentially serve as a docking site for phospholipase D (PLD) after ethylene stimulation. It is hypothesized that ethylene stimulation may activate PI3K resulting in enhanced level of phosphorylated phosphatidylinositol. However, the properties and function of PI3K is not well understood in plants. In the present study, the effect of PI3K inhibition during tomato fruit ripening was evaluated. This study demonstrated that PI3K activity is required for normal ripening process of the fruit. Inhibition of PI3K activity using wortmannin significantly reduced tomato ripening process.
    [Show full text]
  • Structure-Function Analysis of the Catalytic Domain of the Histidine Kinase Chea
    Loyola University Chicago Loyola eCommons Dissertations Theses and Dissertations 1997 Structure-Function Analysis of the Catalytic Domain of the Histidine Kinase Chea Dolph David Ellefson Loyola University Chicago Follow this and additional works at: https://ecommons.luc.edu/luc_diss Part of the Microbiology Commons Recommended Citation Ellefson, Dolph David, "Structure-Function Analysis of the Catalytic Domain of the Histidine Kinase Chea" (1997). Dissertations. 3425. https://ecommons.luc.edu/luc_diss/3425 This Dissertation is brought to you for free and open access by the Theses and Dissertations at Loyola eCommons. It has been accepted for inclusion in Dissertations by an authorized administrator of Loyola eCommons. For more information, please contact [email protected]. This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License. Copyright © 1997 Dolph David Ellefson LOYOLA UNIVERSITY MEDICAL CENTER LIBRARY LOYOLA UNIVERSITY OF CHICAGO STRUCTURE-FUNCTION ANALYSIS OF THE CATALYTIC DOMAIN OF THE HISTIDINE KINASE CHEA A DISSERTATION SUBMITTED TO THE FACULTY OF THE GRADUATE SCHOOL IN CANDIDACY FOR THE DEGREE OF DOCTOR OF PHILOSOPHY DEPARTMENT OF MICROBIOLOGY AND IMMUNOLOGY BY DOLPH DAVID ELLEFSON CHICAGO, ILLINOIS MAY, 1997 Copyright by Dolph David Ellefson, 1997 All Rights Reserved ii ACKNOWLEDGEMENTS I would like to thank my director, Dr. Alan J. Wolfe, for his support, advice, and encouragment during the many years in his laboratory. In his laboratory, I was given a rare opportunity to explore a new arena of science and interact with a field of gifted researchers who I would not known otherwise. I would also like to thank the members of my committee, Ors.
    [Show full text]
  • Targeting the Tryptophan Hydroxylase 2 Gene for Functional Analysis in Mice and Serotonergic Differentiation of Embryonic Stem Cells
    TARGETING THE TRYPTOPHAN HYDROXYLASE 2 GENE FOR FUNCTIONAL ANALYSIS IN MICE AND SEROTONERGIC DIFFERENTIATION OF EMBRYONIC STEM CELLS Inaugural-Dissertation to obtain the academic degree Doctor rerum naturalium (Dr. rer. nat.) submitted to the Department of Biology, Chemistry and Pharmacy of Freie Universität Berlin by Dana Kikic, M.Sc. in Molecular biology and Physiology from Nis June, 2009 The doctorate studies were performed in the research group of Prof. Michael Bader Molecular Biology of Peptide Hormones at Max-Delbrück-Center for Molecular Medicine in Berlin, Buch Mai 2005 - September 2008. 1st Reviewer: Prof. Michael Bader 2nd Reviewer: Prof. Udo Heinemann date of defence: 13. August 2009 ACKNOWLEDGMENTS Herewith, I would like to acknowledge the persons who made this thesis possible and without whom my initiation in the world of basic science research would not have the spin it has now, neither would my scientific illiteracy get the chance to eradicate. I am expressing my very personal gratitude and recognition to: Prof. Michael Bader, for an inexhaustible guidance in all the matters arising during the course of scientific work, for an instinct in defining and following the intellectual challenge and for letting me following my own, for necessary financial support, for defining the borders of reasonable and unreasonable, for an invaluable time and patience, and an amazing efficiency in supporting, motivating, reading, correcting and shaping my scientific language during the last four years. Prof. Harald Saumweber and Prof. Udo Heinemann, for taking over the academic supervision of the thesis, and for breathing in it a life outside the laboratory walls and their personal signature.
    [Show full text]
  • Pseudonocardia Pini Sp. Nov., an Endophytic Actinobacterium Isolated from Roots of the Pine Tree Callitris Preissii
    Pseudonocardia Pini Sp. Nov., An Endophytic Actinobacterium Isolated From Roots of the Pine Tree Callitris Preissii Onuma Kaewkla ( [email protected] ) Mahasarakham University https://orcid.org/0000-0001-7630-7074 Christopher Milton Mathew Franco Flinders University of South Australia: Flinders University Research Article Keywords: Pseudonocardia pini sp. nov., an endophytic actinobacterium Posted Date: March 16th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-274242/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Version of Record: A version of this preprint was published at Archives of Microbiology on April 23rd, 2021. See the published version at https://doi.org/10.1007/s00203-021-02309-3. Page 1/16 Abstract A Gram positive, aerobic, actinobacterial strain with rod-shaped spores, CAP47RT, which was isolated from the surface-sterilized root of a native pine tree (Callitris preissii), South Australia is described. The major cellular fatty acid of this strain was iso-H-C16:1 and major menaquinone was MK-8(H4). The diagnostic diamino acid in the cell-wall peptidoglycan was identied as meso- diaminopimelic acid. These chemotaxonomic data conrmed the aliation of strain CAP47RT to the genus Pseudonocardia. Phylogenetic evaluation based on 16S rRNA gene sequence analysis placed this strain in the family Pseudonocardiaceae, being most closely related to Pseudonocardia xishanensis JCM 17906T (98.8%), Pseudonocardia oroxyli DSM 44984T (98.7%), Pseudonocardia thailandensis CMU-NKS-70T (98.7%), and Pseudonocardia ailaonensis DSM 44979T (97.9%). The results of the polyphasic study which contain genome comparisons of ANIb, ANIm and digital DNA-DNA hybridization revealed the differentiation of strain CAP47RT from the closest species with validated names.
    [Show full text]
  • Functional Analysis of Chimeric Lysin Motif Domain Receptors Mediating Nod Factor-Induced Defense Signaling in Arabidopsis Thali
    The Plant Journal (2014) 78, 56–69 doi: 10.1111/tpj.12450 Functional analysis of chimeric lysin motif domain receptors mediating Nod factor-induced defense signaling in Arabidopsis thaliana and chitin-induced nodulation signaling in Lotus japonicus Wei Wang1,2, Zhi-Ping Xie1,* and Christian Staehelin1,* 1State Key Laboratory of Biocontrol and Guangdong Key Laboratory of Plant Resources, School of Life Sciences, Sun Yat-sen University, East Campus, Guangzhou 510006, China, and 2Anhui Key Laboratory of Plant Genetics & Breeding, School of Life Sciences, Anhui Agricultural University, 130 Changjiang West Road, Hefei, Anhui 230036, China Received 12 October 2013; revised 11 January 2014; accepted 16 January 2014; published online 8 February 2014. *For correspondence (e-mails [email protected] or [email protected]). SUMMARY The expression of chimeric receptors in plants is a way to activate specific signaling pathways by corre- sponding signal molecules. Defense signaling induced by chitin from pathogens and nodulation signaling of legumes induced by rhizobial Nod factors (NFs) depend on receptors with extracellular lysin motif (LysM) domains. Here, we constructed chimeras by replacing the ectodomain of chitin elicitor receptor kinase 1 (AtCERK1) of Arabidopsis thaliana with ectodomains of NF receptors of Lotus japonicus (LjNFR1 and LjNFR5). The hybrid constructs, named LjNFR1–AtCERK1 and LjNFR5–AtCERK1, were expressed in cerk1-2, an A. thaliana CERK1 mutant lacking chitin-induced defense signaling. When treated with NFs from Rhizobi- um sp. NGR234, cerk1-2 expressing both chimeras accumulated reactive oxygen species, expressed chitin- responsive defense genes and showed increased resistance to Fusarium oxysporum. In contrast, expression of a single chimera showed no effects.
    [Show full text]
  • (12) United States Patent DX
    USOO7300.953B2 (12) United States Patent (10) Patent No.: US 7,300,953 B2 Nishino et al. (45) Date of Patent: Nov. 27, 2007 (54) PROCESS FOR PREPARING NITRILE JP 54-122220 A 9, 1979 COMPOUND, CARBOXYLIC ACID JP 59-51251. A 3, 1984 COMPOUND OR CARBOXYLIC ACID JP 8-5O1299. A 2, 1996 ESTER COMPOUND JP 200O281672 A * 10, 2000 WO WO-94/05639 A1 3, 1994 (75) Inventors: Shigeyoshi Nishino, Ube (JP); Kenji Hirotsu, Ube (JP); Hidetaka Shima, OTHER PUBLICATIONS Ube (JP); Keiji Iwamoto, Ube (JP); Lawerence I. Kruse et al., J. Med. Chem. 1990, vol. 33, No. 2, pp. Takashi Harada, Ube (JP) T81 to 789. Christoph Strassier et al., Helvetica Chimica Acta, vol. 80, pp. 1528 (73) Assignee: Ube Industries, Ltd, Ube-shi (JP) to 1554, 1997. (*) Notice: Subject to any disclaimer, the term of this * cited by examiner patent is extended or adjusted under 35 Primary Examiner Kamal A. Saeed U.S.C. 154(b) by 0 days. Assistant Examiner Shawquia Young (74) Attorney, Agent, or Firm—Birch, Stewart, Kolasch & (21) Appl. No.: 10/572,373 Birch, LLP (22) PCT Filed: Sep. 17, 2004 (57) ABSTRACT PCT/UP2004/O13626 (86). PCT No.: The present invention discloses a process for preparing a S 371 (c)(1), nitrile compound, a carboxylic acid compound or a carboxy (2), (4) Date: Mar. 16, 2006 lic acid ester compound represented by the formula (2): (87) PCT Pub. No.: WO2005/0284.10 (2) PCT Pub. Date: Mar. 31, 2005 R (65) Prior Publication Data R 1. R2 US 2006/0287541 A1 Dec.
    [Show full text]
  • Enzyme DHRS7
    Toward the identification of a function of the “orphan” enzyme DHRS7 Inauguraldissertation zur Erlangung der Würde eines Doktors der Philosophie vorgelegt der Philosophisch-Naturwissenschaftlichen Fakultät der Universität Basel von Selene Araya, aus Lugano, Tessin Basel, 2018 Originaldokument gespeichert auf dem Dokumentenserver der Universität Basel edoc.unibas.ch Genehmigt von der Philosophisch-Naturwissenschaftlichen Fakultät auf Antrag von Prof. Dr. Alex Odermatt (Fakultätsverantwortlicher) und Prof. Dr. Michael Arand (Korreferent) Basel, den 26.6.2018 ________________________ Dekan Prof. Dr. Martin Spiess I. List of Abbreviations 3α/βAdiol 3α/β-Androstanediol (5α-Androstane-3α/β,17β-diol) 3α/βHSD 3α/β-hydroxysteroid dehydrogenase 17β-HSD 17β-Hydroxysteroid Dehydrogenase 17αOHProg 17α-Hydroxyprogesterone 20α/βOHProg 20α/β-Hydroxyprogesterone 17α,20α/βdiOHProg 20α/βdihydroxyprogesterone ADT Androgen deprivation therapy ANOVA Analysis of variance AR Androgen Receptor AKR Aldo-Keto Reductase ATCC American Type Culture Collection CAM Cell Adhesion Molecule CYP Cytochrome P450 CBR1 Carbonyl reductase 1 CRPC Castration resistant prostate cancer Ct-value Cycle threshold-value DHRS7 (B/C) Dehydrogenase/Reductase Short Chain Dehydrogenase Family Member 7 (B/C) DHEA Dehydroepiandrosterone DHP Dehydroprogesterone DHT 5α-Dihydrotestosterone DMEM Dulbecco's Modified Eagle's Medium DMSO Dimethyl Sulfoxide DTT Dithiothreitol E1 Estrone E2 Estradiol ECM Extracellular Membrane EDTA Ethylenediaminetetraacetic acid EMT Epithelial-mesenchymal transition ER Endoplasmic Reticulum ERα/β Estrogen Receptor α/β FBS Fetal Bovine Serum 3 FDR False discovery rate FGF Fibroblast growth factor HEPES 4-(2-Hydroxyethyl)-1-Piperazineethanesulfonic Acid HMDB Human Metabolome Database HPLC High Performance Liquid Chromatography HSD Hydroxysteroid Dehydrogenase IC50 Half-Maximal Inhibitory Concentration LNCaP Lymph node carcinoma of the prostate mRNA Messenger Ribonucleic Acid n.d.
    [Show full text]
  • Cationic Oligomerization of Ethylene Oxide
    Polymer Journal, Vol. 15, No. 12, pp 883-889 (1983) Cationic Oligomerization of Ethylene Oxide Shiro KOBAYASHI, Takatoshi KOBAYASHI, and Takeo SAEGUSA Department of Synthetic Chemistry, Faculty of Engineering, Kyoto University, Kyoto 606, Japan (Received July 29, 1983) ABSTRACT: The oligomerization of ethylene oxide (EO) was investigated with several cationic catalysts to find a method for producing cyclic oligomers (crown ethers). The reactions were monitored by NMR spectroscopy (1 H and 19F) and gas chromatography. The composition of oligomers was found to change during the reactions, and the production of 1,4-dioxane increased at the later stage of the reactions. This indicates that the composition of oligomers is kinetically controlled. The formation of higher cyclic oligomers was favored by the addition of tetrahydro­ pyran or 1,4-dioxane to the system catalyzed by an oxonium salt; the maximum yield of cyclic tetramer was 16.8%. The effect of alkali metal salts was also examined. A template effect was observed to increase the amount of cyclic oligomer production. KEY WORDS Cationic Oligomerization I Ethylene Oxide I Cyclic Oligomers I Crown Ether I Kinetically Controlled Reaction I Additive Effects of Metal Salts I Template Effect I An extensive study was recently carried out on EXPERIMENTAL the cationic. polymerization of heterocyclic mono­ mers which, it was found, may lead to polymers Materials containing significant amounts of linear and cyclic All reagents were distilled under nitrogen. A oligomers.1 The most thoroughly studied monomer commercial sample of EO was distilled twice. was ethylene oxide (EO). Eastham et a!. observed Tetrahydropyran (THP) and DON were dried with that the cationic polymerization of EO resulted in sodium metal and distilled.
    [Show full text]