Functional Roles of Each of the 400 Putative Gene Products Used for Inferring Bradyrhizobium Phylogeny
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Symphytum Officinale) Hairy Roots by Rnai Silencing of Homospermidine Synthase
Published online: 2019-08-26 Original Papers Reduction of Pyrrolizidine Alkaloid Levels in Comfrey (Symphytum officinale) Hairy Roots by RNAi Silencing of Homospermidine Synthase Authors Lars H. Kruse 1, 3, Thomas Stegemann1, Julia Jensen-Kroll 1, Annika Engelhardt 1, Anne-Maria Wesseling 1, Annemarie Lippert 2, Jutta Ludwig-Müller 2,DietrichOber1 Affiliations ABSTRACT 1 Botanisches Institut, Kiel University, Kiel, Germany Comfrey is a medicinal plant, extracts of which are tradition- 2 Institut für Botanik, Technische Universität Dresden, ally used for the treatment of painful inflammatory muscle Dresden, Germany and joint problems, because the plant contains allantoin and 3 Plant Biology Section, School of Integrative Plant Science, rosmarinic acid. However, its medicinal use is limited because Cornell University, USA of its toxic pyrrolizidine alkaloid (PA) content. PAs encompass more than 400 different compounds that have been identified Key words from various plant lineages. To date, only the first pathway- Symphytum officinale, alkaloid extraction, transgenic tissue specific enzyme, homospermidine synthase (HSS), has been culture, transcript quantification, Boraginaceae characterized. HSS catalyzes the formation of homospermi- dine, which is exclusively incorporated into PAs. HSS has been received May 6, 2019 recruited several times independently in various plant line- revised August 5, 2019 ages during evolution by duplication of the gene encoding de- accepted August 15, 2019 oxyhypusine synthase (DHS), an enzyme of primary metabo- Bibliography lism. Here, we describe the establishment of RNAi knockdown DOI https://doi.org/10.1055/a-0998-5125 hairy root mutants of HSS in Symphytum officinale. A knock- – Published online August 26, 2019 | Planta Med 2019; 85: down of HSS by 60 80% resulted in a significant reduction of 1177–1186 © Georg Thieme Verlag KG Stuttgart · New York | homospermidine by ~ 86% and of the major PA components ‑ ‑ ISSN 0032‑0943 7 acetylintermedine N-oxide and 3 acetylmyoscorpine N-ox- ide by approximately 60%. -
Pyrrolizidine Alkaloids: Biosynthesis, Biological Activities and Occurrence in Crop Plants
molecules Review Pyrrolizidine Alkaloids: Biosynthesis, Biological Activities and Occurrence in Crop Plants Sebastian Schramm, Nikolai Köhler and Wilfried Rozhon * Biotechnology of Horticultural Crops, TUM School of Life Sciences Weihenstephan, Technical University of Munich, Liesel-Beckmann-Straße 1, 85354 Freising, Germany; [email protected] (S.S.); [email protected] (N.K.) * Correspondence: [email protected]; Tel.: +49-8161-71-2023 Academic Editor: John C. D’Auria Received: 20 December 2018; Accepted: 29 January 2019; Published: 30 January 2019 Abstract: Pyrrolizidine alkaloids (PAs) are heterocyclic secondary metabolites with a typical pyrrolizidine motif predominantly produced by plants as defense chemicals against herbivores. They display a wide structural diversity and occur in a vast number of species with novel structures and occurrences continuously being discovered. These alkaloids exhibit strong hepatotoxic, genotoxic, cytotoxic, tumorigenic, and neurotoxic activities, and thereby pose a serious threat to the health of humans since they are known contaminants of foods including grain, milk, honey, and eggs, as well as plant derived pharmaceuticals and food supplements. Livestock and fodder can be affected due to PA-containing plants on pastures and fields. Despite their importance as toxic contaminants of agricultural products, there is limited knowledge about their biosynthesis. While the intermediates were well defined by feeding experiments, only one enzyme involved in PA biosynthesis has been characterized so far, the homospermidine synthase catalyzing the first committed step in PA biosynthesis. This review gives an overview about structural diversity of PAs, biosynthetic pathways of necine base, and necic acid formation and how PA accumulation is regulated. Furthermore, we discuss their role in plant ecology and their modes of toxicity towards humans and animals. -
REVIEW ARTICLES Pyrrolizidine Alkaloids
Review articles Pyrrolizidine alkaloids – chemistry, biosynthesis, pathway, toxicity, safety and perspectives of medicinal usage Mariola DREGER, MARZENA Stanisławska, ANNA Krajewska-Patan, SEBASTIAN Mielcarek, Przemysław Łukasz Mikołajczak, WALDEMAR Buchwald The Branch of Medicinal Plants Institute of Natural Fibres and Medicinal Plants Libelta 27 61-707 Poznań, Poland *corresponding author: [email protected] S u m m a r y Pyrrolizidine alkaloids (PAs) are the class of secondary metabolites that evolved as a powerful tool in the plant defensive interactions against herbivores. The occurrence of PAs in the plant world is scattered in several unrelated botanic families with special abun- dance in Asteraceae, Boraginaceae and Fabaceae. Homospermidine synthase (HSS) was recognized as a key enzyme that catalyzes homospermidine formation from polyamines. The studies of HSS kinetic and gene sequence revealed that it is of polyphyletic origin and raised as a result of deoxyhypusine synthase (DHS) gene duplication. The ability of PAs production occurred independently at least four times in course of plant evolution. The PAs biosynthesis is tightly correlated with growth phase and biomass production. It is sup- posed that PAs biosynthesis is individually regulated in different lineages of plants. The PAs with a 1,2 unsaturated necine skeleton show toxic activity (hepatoxicity, carcinogenicity, genotoxicity, teratogenocity and cytotoxicity). It is though that pyrrolic esters formation during the detoxication process in the liver is the main mechanism of PAs toxicity. The pyr- rolic esters are highly reactive and tend to bind rapidly with nucleophilic macromolecules including DNA and DNA-protein inducing hepatotoxicity or tumorigenecity. The problem of PAs toxicity cause the restrictions in the production and sale of herbal products. -
University of Florida Thesis Or Dissertation Formatting Template
METABOLOMIC CHARACTERIZATION OF THE INTESTINAL BACTERIAL OXALOBIOME By CASEY A. CHAMBERLAIN A DISSERTATION PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY UNIVERSITY OF FLORIDA 2019 © 2019 Casey A. Chamberlain To my wife, Michelle ACKNOWLEDGMENTS Obtaining a PhD may only bestow the title of “Doctor” upon one individual, but the journey to such an achievement is often aided by, and impossible without, the loving support of family, friends, and others along the way. I am thankful for the Lord for His providence and love and for blessing me with the unique interests, talents, and gifts that gave me the opportunity to fulfill this achievement. I am eternally grateful for my wife, Michelle, for providing me with love, support, comfort, and guidance, which propagates confidence, happiness, and fulfillment into all areas of my life. Her guiding and strengthening example has molded me into the man I am today. I am blessed to have my daughter, Maci, who reminds me daily of the purpose of this life and the love the Lord feels for each of His children. I am fortunate to have received love and support from family, both immediate and extended, which helps foster a sense of wholeness in my life. I am uplifted and reinforced by the friends I have made during this season of life, both inside and outside the lab, particularly those whom I have come to know closely on a personal level. I respect, honor, and am indebted to Tim Garrett for providing me the opportunity for this journey. -
Bill Ceevee 2014
curriculum vitae, William F. Martin Institutional address: Home address: Institut für Botanik III Rilkestrasse 13 Heinrich Heine-Universität Düsseldorf D-41469 Neuss Universitätsstraße 1 Germany D-40225 Düsseldorf Germany e-mail: [email protected] Tel. +49-211-811-3011 Date of birth : 16.02.57 in Bethesda, Maryland, USA Familial status : Married, four children Nationality : USA University degree : 1981-1985, Technische Universität Hannover, Germany: Biology Diplom thesis : 1985, Institut für Botanik, TU Hannover with Rüdiger Cerff: Botany PhD thesis : 1985-1988, Max-Planck-Institut für Züchtungsforschung, Cologne, with Heinz Saedler; degree conferred by the University of Cologne: Genetics Postdoc : 1988-1989, Max-Planck-Institut für Züchtungsforschung, Cologne Postdoc : 1989-1999, Institut für Genetik, Technische Universität Braunschweig Habilitation : 1992, TU Braunschweig, Germany, venia legendi for the field of Botany Full professor : 1999-2011 for "Ecological Plant Physiology" (C4), Universität Düsseldorf : 2011- for "Molecular Evolution" (C4), Universität Düsseldorf Honours 2013- Visiting Professor, Instituto de Tecnologia Química e Biológica, Oeiras, Portugal 2012 Elected Member of EMBO (European Molecular Biology Organisation) 2010- Science Advisory Committe, Helmholz Alliance Planetry Evoution and Life 2008 Elected Member of the Nordrhein-Westfälische Akademie der Wissenschaften 2007-2012 Selection Committee for the Heinz-Maier-Leibnitz Prize of the DFG 2006 Elected Fellow of the American Academy for Microbiology 2006-2009 Julius von -
Supplementary Material Toxicological Impacts and Likely Protein Targets Of
Supplementary Material Toxicological impacts and likely protein targets of bisphenol A in Paramecium caudatum Marcus V. X. Senra† & Ana Lúcia Fonseca Instituto de Recursos Naturais, Universidade Federal de Itajubá, 37500-903, Itajubá, Minas Gerais – Brazil †To whom correspondence should be addressed – [email protected]; Orcid - 0000-0002-3866- 8837 Table S1. Annotation data on the P. caudatum 3D modelled proteins and their binding energies to BPA. BINDING ID DESCRIPTION CHROMOSOME NT_START NT_END ENERGIES (kcal/mol) PCAU.43c3d.1.P00010012 Tryptophan synthase beta subunit-like PLP-dependent enzyme scaffold_0001 23197 24853 -7.4 PCAU.43c3d.1.P00010015 Ribosomal protein L32e scaffold_0001 26373 26859 -6.2 PCAU.43c3d.1.P00010044 Catalase, mono-functional, haem-containing scaffold_0001 71821 73367 -6.5 PCAU.43c3d.1.P00010050 Dihydroorotate dehydrogenase, class 1/ 2 scaffold_0001 76614 79650 -6.6 PCAU.43c3d.1.P00010054 Serine/threonine/dual specificity protein kinase, catalytic domain scaffold_0001 83399 84653 -6.7 PCAU.43c3d.1.P00010070 Peptidyl-prolyl cis-trans isomerase, FKBP-type scaffold_0001 104909 105387 -5.9 PCAU.43c3d.1.P00010103 V-ATPase proteolipid subunit C-like domain scaffold_0001 168736 169346 -5.6 PCAU.43c3d.1.P00010112 DNA-directed RNA polymerase, RBP11-like dimerisation domain scaffold_0001 180310 181372 -6.0 PCAU.43c3d.1.P00010165 Vacuolar (H+)-ATPase G subunit scaffold_0001 252653 253112 -5.6 PCAU.43c3d.1.P00010176 Coproporphyrinogen III oxidase, aerobic scaffold_0001 262051 263168 -6.7 PCAU.43c3d.1.P00010205 Metalloenzyme, -
Biosynthesis and Regulation of Polyamines in Higher Plants
Plant Growth Regulation 3:205-226 H985). [3] © 1985 Martinus Ni/hoff/Dr W. Junk Publishers, Dordrecht. Pn'nted in the Netherlands. Biosynthesis and regulation of polyamines in higher plants P.R. ADIGA and G.L. PRASAD Department of Biochemistry, Indian Institute of Science, Bangalore 560 012, India (Received 10 Oct. 1984; accepted 15 Oct. 1984) 1. Introduction The polyamines (PAs), spermidine (SPD) and spermine (SPN) and their diamine precursor, putrescine (PUT) represent a set of evolutionarily highly conserved small molecular weight organic polycations which play vital roles as modulators of a plethora of biological processes from enzyme activation and maintenance of ionic balance, through regulation of growth and development, to mediation of hormone action and progress of cell division cycle. Because of this functional versatility, research on PAs represents one of the most vigorously pursued areas of modern biology. While most of the currently available information on these important classes of bioregulators is derived from microbial and animal systems, interest in their possible participation in various facets of plant biochemistry and physiology is of relatively recent origin. The early pioneering investigations of Smith and others [34, 48, 62, 54, 63] on K+-deficient barley leaves, set the stage for the elucidation of the general metabolic sequences involved in the biogenesis of these amines in plants and pinpointed the important roles PAs play in maintenance of intracellular pH and ionic balance. How- ever, with the recognition that plant systems also offer tremendous potential for unravelling the riddles concerned with the biological functions of PAs, new vistas have been opened in plant PA research. -
12) United States Patent (10
US007635572B2 (12) UnitedO States Patent (10) Patent No.: US 7,635,572 B2 Zhou et al. (45) Date of Patent: Dec. 22, 2009 (54) METHODS FOR CONDUCTING ASSAYS FOR 5,506,121 A 4/1996 Skerra et al. ENZYME ACTIVITY ON PROTEIN 5,510,270 A 4/1996 Fodor et al. MICROARRAYS 5,512,492 A 4/1996 Herron et al. 5,516,635 A 5/1996 Ekins et al. (75) Inventors: Fang X. Zhou, New Haven, CT (US); 5,532,128 A 7/1996 Eggers Barry Schweitzer, Cheshire, CT (US) 5,538,897 A 7/1996 Yates, III et al. s s 5,541,070 A 7/1996 Kauvar (73) Assignee: Life Technologies Corporation, .. S.E. al Carlsbad, CA (US) 5,585,069 A 12/1996 Zanzucchi et al. 5,585,639 A 12/1996 Dorsel et al. (*) Notice: Subject to any disclaimer, the term of this 5,593,838 A 1/1997 Zanzucchi et al. patent is extended or adjusted under 35 5,605,662 A 2f1997 Heller et al. U.S.C. 154(b) by 0 days. 5,620,850 A 4/1997 Bamdad et al. 5,624,711 A 4/1997 Sundberg et al. (21) Appl. No.: 10/865,431 5,627,369 A 5/1997 Vestal et al. 5,629,213 A 5/1997 Kornguth et al. (22) Filed: Jun. 9, 2004 (Continued) (65) Prior Publication Data FOREIGN PATENT DOCUMENTS US 2005/O118665 A1 Jun. 2, 2005 EP 596421 10, 1993 EP 0619321 12/1994 (51) Int. Cl. EP O664452 7, 1995 CI2O 1/50 (2006.01) EP O818467 1, 1998 (52) U.S. -
Homospermidine, Spermidine, and Putrescine: The
AN ABSTRACT OF THE THESIS OF Paula Allene Tower for the degree of Doctor of Philosophy in Microbiology presented on July 28, 1987. Title: Homospermidine, Spermidine, and Putrescine: The Biosynthesis and Metabolism of Polyamines in Rhizobium meliloti Redacted for privacy Abstract approved: Dr.4dolph J. Ferro Rhizobium, in symbiotic association with leguminous plants, is able to fix atmospheric nitrogen after first forming root nodules. Since polyamines are associated with and found in high concentration in rapidly growing cells and are thought to be important foroptimal cell growth, it is possible that these polycations are involved in the extensive cell proliferation characteristic of the nodulation process. As a first step towards elucidating the role(s) of poly- amines in the rhizobial-legume interaction, I have characterized polyamine biosynthesis and metabolism in free-living Rhizobium meliloti. In addition to the detection of putrescine and spermidine, the presence of a less common polyamine, homospermidine, wasconfirmed in Rhizobium meliloti, with homospermidine comprising 79 percent of the free polyamines in this procaryote. The presence of an exogenous polyamine both affected the intra- cellular levels of each polyamine pool and inhibited the accumula- tion of a second amine. DL-a-Difluoromethylornithine (DFMO), an irreversible inhibitor of ornithine decarboxylase, wasfound to: (1) inhibit the rhizobial enzyme both in vitro and in vivo; (2) increase the final optical density; and (3) create ashift in the dominant polyamine from homospermidine to spermidine. A series of radiolabeled amino acids and polyamines were studied as polyamine precursors. Ornithine, arginine, aspartic acid, putrescine, and spermidine, but not methionine, resulted in the isolation of labeled putrescine, spermidine, andhomospermidine. -
Supplementary Figure 1. Rpod Sequence Alignment. Protein Sequence Alignment Was Performed for Rpod from Three Species, Pseudoruegeria Sp
J. Microbiol. Biotechnol. https://doi.org/10.4014/jmb.1911.11006 J. Microbiol. Biotechnol. https://doi.org/10.4014/jmb.2003.03025 Supplementary Figure 1. RpoD sequence alignment. Protein sequence alignment was performed for RpoD from three species, Pseudoruegeria sp. M32A2M (FPS10_24745, this study), Ruegeria pomeroyi (NCBI RefSeq: WP_011047484.1), and Escherichia coli (NCBI RefSeq: NP_417539.1). Multalin version 5.4.1 was used for the analysis. Subregion 2 and 4 are represented in a rectangle, and the helix-turn-helix motif in subregion 4 is highlighted in red. The amino acid degeneration from any of the three was highlighted in light gray and the sequence degeneration between Pseudoruegeria sp. M32A2M and R. pomeroyi is highlighted in dark gray. The substituted two amino acids into HTH motif (K578Q and D581S) against E. coli were marked as asterisk. Supplementary Table 1. Genome assembly statistics Categories Pseudoruegeria sp. M32A2M Number of scaffolds less than 1,000 bp 0 Number of scaffolds between 1,000 bp –10,000 bp 39 Number of scaffolds between 10,000 bp – 100,000 bp 38 Number of scaffolds larger than 100,000 bp 14 Number of scaffolds 91 Total assembled length (bp) 5,466,515 G+C contents (%) 62.4 N50 (bp) 249,384 Minimum length of scaffold (bp) 1,015 Maximum length of scaffold (bp) 733,566 Total Ns included in the draft genome 2,158 Supplementary Table 2. The list of gene annotation and its functional categorization in Pseudoruegeria sp. -
Cloning and Expression of Deoxyhypusine Synthase from Plasmodium Vivax and Theileria Parva As an Approach for Target Evaluation in Anti-Parasitic Chemotherapy
Cloning and expression of deoxyhypusine synthase from Plasmodium vivax and Theileria parva as an approach for target evaluation in anti-parasitic chemotherapy Dissertation zur Erlangung des Doktorgrades (Dr. rer. nat.) der Mathematisch-Naturwissenschaftlichen Fakultät der Rheinischen Friedrich-Wilhelms-Universität Bonn vorgelegt von James Thuo Njuguna aus Ol Kalou,Kenya Bonn (2009) I Angefertigt mit Genehmigung der Mathematisch-Naturwissenschaftlichen Fakultät der Rheinischen Friedrich-Wilhelms-Universität Bonn 1. Gutachter: Prof. Dr. Annette.E. Kaiser 2. Gutachter: Prof. Dr. Christa E. Müller Tag der Promotion: (31-09-2009) II Acknowledgments This thesis has come to realization because of the support and mentoring of my supervisors Prof. Dr. Annette Kaiser and Prof. Dr Christa Mueller. Prof. Dr Kaiser led me to this area of study and has kept me focused on our scientific goals even when faced with apparently insurmountable challenges. My co-supervisor Prof. Dr. Prof. Christa Mueller has been of great help and support during the last phase of the study. To both I express my deepest gratitude. This Thesis was only possible because of the logistical support by Peter Croll and his institute the Bonn International Center for Conversion (BICC); I will always be indebted to him for this support and for his ideas on positive human development. I am very thankful to Prof. Dr. Achim Hörauf for hosting my study at the Institute for Medical Microbiology, Immunology and Parasitology when I first got started and for his continued interest in our progress. I would like to also thank Prof. Dr. U. Holzgrabe for availing to me the piperidones evaluated in this Thesis, Dr. -
(12) Patent Application Publication (10) Pub. No.: US 2012/0266329 A1 Mathur Et Al
US 2012026.6329A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2012/0266329 A1 Mathur et al. (43) Pub. Date: Oct. 18, 2012 (54) NUCLEICACIDS AND PROTEINS AND CI2N 9/10 (2006.01) METHODS FOR MAKING AND USING THEMI CI2N 9/24 (2006.01) CI2N 9/02 (2006.01) (75) Inventors: Eric J. Mathur, Carlsbad, CA CI2N 9/06 (2006.01) (US); Cathy Chang, San Marcos, CI2P 2L/02 (2006.01) CA (US) CI2O I/04 (2006.01) CI2N 9/96 (2006.01) (73) Assignee: BP Corporation North America CI2N 5/82 (2006.01) Inc., Houston, TX (US) CI2N 15/53 (2006.01) CI2N IS/54 (2006.01) CI2N 15/57 2006.O1 (22) Filed: Feb. 20, 2012 CI2N IS/60 308: Related U.S. Application Data EN f :08: (62) Division of application No. 1 1/817,403, filed on May AOIH 5/00 (2006.01) 7, 2008, now Pat. No. 8,119,385, filed as application AOIH 5/10 (2006.01) No. PCT/US2006/007642 on Mar. 3, 2006. C07K I4/00 (2006.01) CI2N IS/II (2006.01) (60) Provisional application No. 60/658,984, filed on Mar. AOIH I/06 (2006.01) 4, 2005. CI2N 15/63 (2006.01) Publication Classification (52) U.S. Cl. ................... 800/293; 435/320.1; 435/252.3: 435/325; 435/254.11: 435/254.2:435/348; (51) Int. Cl. 435/419; 435/195; 435/196; 435/198: 435/233; CI2N 15/52 (2006.01) 435/201:435/232; 435/208; 435/227; 435/193; CI2N 15/85 (2006.01) 435/200; 435/189: 435/191: 435/69.1; 435/34; CI2N 5/86 (2006.01) 435/188:536/23.2; 435/468; 800/298; 800/320; CI2N 15/867 (2006.01) 800/317.2: 800/317.4: 800/320.3: 800/306; CI2N 5/864 (2006.01) 800/312 800/320.2: 800/317.3; 800/322; CI2N 5/8 (2006.01) 800/320.1; 530/350, 536/23.1: 800/278; 800/294 CI2N I/2 (2006.01) CI2N 5/10 (2006.01) (57) ABSTRACT CI2N L/15 (2006.01) CI2N I/19 (2006.01) The invention provides polypeptides, including enzymes, CI2N 9/14 (2006.01) structural proteins and binding proteins, polynucleotides CI2N 9/16 (2006.01) encoding these polypeptides, and methods of making and CI2N 9/20 (2006.01) using these polynucleotides and polypeptides.