DOCSLIB.ORG

(12) Patent Application Publication (10) Pub. No.: US 2016/0186168 A1 Konieczka Et Al

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
(12) Patent Application Publication (10) Pub. No.: US 2016/0186168 A1 Konieczka Et Al US 2016O1861 68A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2016/0186168 A1 Konieczka et al. (43) Pub. Date: Jun. 30, 2016 (54) PROCESSES AND HOST CELLS FOR Related U.S. Application Data GENOME, PATHWAY. AND BIOMOLECULAR (60) Provisional application No. 61/938,933, filed on Feb. ENGINEERING 12, 2014, provisional application No. 61/935,265, - - - filed on Feb. 3, 2014, provisional application No. (71) Applicant: ENEVOLV, INC., Cambridge, MA (US) 61/883,131, filed on Sep. 26, 2013, provisional appli (72) Inventors: Jay H. Konieczka, Cambridge, MA cation No. 61/861,805, filed on Aug. 2, 2013. (US); James E. Spoonamore, Publication Classification Cambridge, MA (US); Ilan N. Wapinski, Cambridge, MA (US); (51) Int. Cl. Farren J. Isaacs, Cambridge, MA (US); CI2N 5/10 (2006.01) Gregory B. Foley, Cambridge, MA (US) CI2N 15/70 (2006.01) CI2N 5/8 (2006.01) (21) Appl. No.: 14/909, 184 (52) U.S. Cl. 1-1. CPC ............ CI2N 15/1082 (2013.01); C12N 15/81 (22) PCT Filed: Aug. 4, 2014 (2013.01); C12N 15/70 (2013.01) (86). PCT No.: PCT/US1.4/49649 (57) ABSTRACT S371 (c)(1), The present disclosure provides compositions and methods (2) Date: Feb. 1, 2016 for genomic engineering. Patent Application Publication Jun. 30, 2016 Sheet 1 of 4 US 2016/O186168 A1 Patent Application Publication Jun. 30, 2016 Sheet 2 of 4 US 2016/O186168 A1 &&&&3&&3&&**??*,º**)..,.: ××××××××××××××××××××-************************** Patent Application Publication Jun. 30, 2016 Sheet 3 of 4 US 2016/O186168 A1 No.vaegwzºkgwaewaeg Patent Application Publication Jun. 30, 2016 Sheet 4 of 4 US 2016/O186168 A1 US 2016/01 86168 A1 Jun. 30, 2016 PROCESSES AND HOST CELLS FOR 0006. In various aspects, the invention is applicable to GENOME, PATHWAY. AND BIOMOLECULAR phenotype engineering, including but not limited to: change ENGINEERING in carbon Substrate utilization, increased cell growth rate, increased production of a desired chemical, redox cofactor PRIORITY balance, reduced production of one or more undesired 0001. This application claims benefit of and priority to byproducts, increased resistance to industrial fermentation, U.S. Provisional Application No. 61/861,805 filed Aug. 2, and increased recombinant protein production, among others. 2013,61/883,131 filed Sep. 26, 2013, 61/935,265 filed Feb.3, In some embodiments, the phenotype relates to pathway engi 2014, and 61/938,933 filed Feb. 12, 2014, each of which is neering, such as where metabolic flux through a core or hereby incorporated by reference in its entirety. primary metabolic pathway is significantly altered (e.g., Sig nificantly altered as compared to wild type strain, an unengi FIELD OF THE INVENTION neered strain, or a starting strain), including alteration of metabolic flux through one or more intermediates that repre 0002. The invention relates to, inter alia, methods and sent metabolic branch points. In various embodiments, the compositions for genome-scale editing of genetic informa invention is applicable to introducing and balancing of heter tion. ologous recombinant enzyme activity with endogenous metabolism, to limit effects on viability and growth, for BACKGROUND example. In some embodiments, the phenotype relates to 0003. Successful genomic and pathway engineering engineering alterations among one or more proteins, such as, requires that metabolic flux be increased through select path by way of illustration, enzymes, for improved biochemical or ways, while not substantially interfering with viability and/or biophysical properties. growth of the organism, or a desired phenotype. This can be 0007. In various aspects, the invention provides methods especially pertinent for substrates or intermediates of the for pathway engineering, in which metabolic flux is altered desired pathway that are involved in core or primary metabo through one or more intermediates of glycolysis, pentose lism, or for branch intermediates involved in more than one phosphate pathway, TCA cycle, one or more secondary bio pathway. Stephanopoulos, Metabolic Fluxes and Metabolic synthesis pathways (e.g., amino acid or nucleotide biosynthe Engineering, Metabolic Engineering 1, 1-11 (1999). In fact, sis pathway), the mevalonate or non-mevalonate pathway, genetic alterations, or combinations of genetic alterations, pathways involved in Sulfur or nitrogen metabolism, and oth that increase metabolic flux through a desired pathway are ers. The invention relates to, in various embodiments, engi difficult to predict, limiting the usefulness of rational engi neering cells to result in one or more of increased or diversi neering approaches. Kern A, et al., Engineering primary fied carbon Substrate utilization, increased or maintained metabolic pathways of industrial microorganisms, J. Biotech growth rate, modified enzyme activity at metabolic branch nology 129: 6-29 (2007). Further, it is often impractical to points, decreased metabolic flux through one or more com generate random and discrete mutational events in vivo and peting secondary pathways and/or increase in flux through a screen or select for improved metabolic flux or biomolecular desired secondary biosynthetic pathway, balanced cellular function. redox chemistry (e.g., redox cofactor balance), balanced het 0004 Methods are needed for screening the genetic space erologous enzyme activity with the endogenous metabolism, of a host organism to identify changes in endogenous genes including reduction in toxic intermediates, increased resis and/or heterologous recombinant genes that provide tance to environmental (e.g., industrial) conditions, increased improved phenotypes, such as in metabolic flux and balance, recombinant protein production, and increased yield of So as to improve or optimize microbial processes, including desired product. production of desired chemicals and biomolecules at indus 0008. In various aspects, the invention engineers cells trial levels, or bioremediation applications. using one or a combination of recombineering systems. The recombineering systems each offer distinct advantages in SUMMARY OF THE INVENTION engineering cells. In some embodiments, the method pro 0005. In various aspects, the invention provides methods duces a library of mutants cells using a ssDNA recombinase for genomic and pathway engineering. The methods are use system, which may include a single-stranded annealing pro ful in E. coli, as well as bacterial cells that are harder to tein (SSAP), such as the W. Red recombineering system (e.g., genetically manipulate but are otherwise valuable for produc Beta protein) or RecET system (e.g., recT), or homologous tion of chemicals, including Bacillus sp. (e.g., Bacillus sub system, which in Some embodiments offers advantages in tilis), Streptomycetes (e.g., Streptomyces avermitilis, Strep identifying pathways, genes, or regions of genes for alter tomyces coelicolor; Streptomyces lividins, Streptomyces ation, as well as targeting specific regions for genetic diver cinnamomensis, Streptomyces collinus, etc.), Cyanobacteria sification. The W. Red operon encodes ssDNA annealing pro (e.g., Synechocystis spp., Prochlorococcus spp., Nostoc tein Beta, which promotes annealing of single stranded punctiforme, Calothrix spp., Aphanizomenon flos-aquae, oligonucleotides at the lagging Strand of the replication fork. Arthrospira platensis, etc.), and Corynebacteria (e.g., The Red recombineering system, as with other SSAP sys Corynebacteria glutamicum, Corynebacteria ammoni tems, may further involve a deletion, inactivation, or reduc agenes, etc.), among others. The methods are further appli tion in mismatch repair (e.g., deletion, inactivation, or reduc cable in Some embodiments to Eukaryotic cells, such as tion in activity or level of mutS). Such a system is useful for yeasts (e.g., Pichia sp., Saccharomyces sp., Schizosaccharo engineering E. coli and other prokaryotic systems, and may myces sp., Kluyveromyces sp., etc.), filamentous fungi (e.g., be engineered into other species described herein. Specifi Neurospora sp. and Aspergillus sp., Penicillium, etc.), algae cally, an oligonucleotide library designed to Screen for ben (e.g., Botryococcus, Chlorella, Dunaliella, Gracilaria, Pleu eficial mutations (or combinations of mutations) in a target rochrysis, and Sargassum, etc.), and plants. genetic 'space' is introduced into the organism, and a desired US 2016/01 86168 A1 Jun. 30, 2016 phenotype identified. The genetic “space' includes identified introduced into the system. Coupling the site-specific pro target pathways (e.g., a primary metabolic pathway and/or grammable nuclease system to the donor sequence on the one or more secondary competing or biosynthetic pathways), same DNA results in a Scarless method for selecting recom target genes, and the nature and diversity of alterations to binants and obviates the need for an additional selectable Screen, including changes to gene regulatory sequences Such marker. Thus, recombination with the homologous target as promoters and transcriptional enhancer sequences, riboso region mutated at the wild-type locus targeted by the site mal binding sites and other sites relating to the efficiency of specific nuclease permits the cell to escape restriction by the transcription, translation, or RNA processing, as well as cod programmed nuclease system. In other words, recombination ing sequence mutations that control the activity, post-transla with the mutated sequence frees the organism from the tional modification, or turnover of the encoded proteins. Oli genomic instability caused by
Load more

Recommended publications
  • Bacteria Belonging to Pseudomonas Typographi Sp. Nov. from the Bark Beetle Ips Typographus Have Genomic Potential to Aid in the Host Ecology
    insects Article Bacteria Belonging to Pseudomonas typographi sp. nov. from the Bark Beetle Ips typographus Have Genomic Potential to Aid in the Host Ecology Ezequiel Peral-Aranega 1,2 , Zaki Saati-Santamaría 1,2 , Miroslav Kolaˇrik 3,4, Raúl Rivas 1,2,5 and Paula García-Fraile 1,2,4,5,* 1 Microbiology and Genetics Department, University of Salamanca, 37007 Salamanca, Spain; [email protected] (E.P.-A.); [email protected] (Z.S.-S.); [email protected] (R.R.) 2 Spanish-Portuguese Institute for Agricultural Research (CIALE), 37185 Salamanca, Spain 3 Department of Botany, Faculty of Science, Charles University, Benátská 2, 128 01 Prague, Czech Republic; [email protected] 4 Laboratory of Fungal Genetics and Metabolism, Institute of Microbiology of the Academy of Sciences of the Czech Republic, 142 20 Prague, Czech Republic 5 Associated Research Unit of Plant-Microorganism Interaction, University of Salamanca-IRNASA-CSIC, 37008 Salamanca, Spain * Correspondence: [email protected] Received: 4 July 2020; Accepted: 1 September 2020; Published: 3 September 2020 Simple Summary: European Bark Beetle (Ips typographus) is a pest that affects dead and weakened spruce trees. Under certain environmental conditions, it has massive outbreaks, resulting in attacks of healthy trees, becoming a forest pest. It has been proposed that the bark beetle’s microbiome plays a key role in the insect’s ecology, providing nutrients, inhibiting pathogens, and degrading tree defense compounds, among other probable traits. During a study of bacterial associates from I. typographus, we isolated three strains identified as Pseudomonas from different beetle life stages. In this work, we aimed to reveal the taxonomic status of these bacterial strains and to sequence and annotate their genomes to mine possible traits related to a role within the bark beetle holobiont.
    [Show full text]
  • 要約) Doctoral Dissertation Antibiotics Shapes Population-Level Diversity In
    ) Doctoral Dissertation Antibiotics shapes population-level diversity in the human gut microbiome ( ) Nishijima Suguru Acknowledgments I would like to express my sincere gratitude to my supervisor, Prof. Masahira Hattori, whose expertise, knowledge and continuous encouragement throughout my research. My sincere thanks also go to Assoc. Prof. Kenshiro Oshima (The University of Tokyo), Dr. Wataru Suda and Dr. Seok-Won Kim for their motivation, immense support and encouragement throughout my work. I am also grateful to all my collaborators, Prof. Hidetoshi Morita (Okayama University) for his fecal sample collection, DNA isolation and sincere encouragement, Prof. Kenya Honda and Dr. Koji Atarashi (Keio University) for mice experiments, Assoc. Prof. Masahiro Umezaki (the University of Tokyo) for support for dietary data analysis, Dr. Todd D. Taylor (RIKEN) for support for writing manuscript and Dr. Yuu Hirose (Toyohashi University of technology) for DNA sequencing. I also would like to thank all past and present members of our laboratory, Erica Iioka, Misa Takagi, Emi Omori, Hiromi Kuroyamagi, Naoko Yamashita, Keiko Komiya, Rina Kurokawa, Chie Shindo, Yukiko Takayama and Yasue Hattori for their great technical support and kind assistance. i Antibiotics shapes population-level diversity in the human gut microbiome () Abstract The human gut microbiome has profound influences on the host’s physiology through its interference with various intestinal functions. The development of next-generation sequencing (NGS) technologies enabled us to comprehensively explore ecological and functional features of the gut microbiomes. Recent studies using the NGS-based metagenomic approaches have suggested high ecological diversity of the microbiome across countries. However, little is known about the structure and feature of the Japanese gut microbiome, and the factor that shapes the population-level diversity in the human gut microbiome.
    [Show full text]
  • Purification, Crystallization and Preliminary X-Ray Diffraction Analysis of Exodeoxyribonuclease III from Crenarchaeon Sulfolobus Tokodaii Strain 7
    Crystal Structure Theory and Applications, 2013, 2, 155-158 Published Online December 2013 (http://www.scirp.org/journal/csta) http://dx.doi.org/10.4236/csta.2013.24021 Purification, Crystallization and Preliminary X-Ray Diffraction Analysis of Exodeoxyribonuclease III from Crenarchaeon Sulfolobus tokodaii Strain 7 Shuichi Miyamoto1*, Chieko Naoe2, Masaru Tsunoda3, Kazuo T. Nakamura2 1Faculty of Pharmaceutical Sciences, Sojo University, Kumamoto, Japan 2School of Pharmacy, Showa University, Tokyo, Japan 3Faculty of Pharmacy, Iwaki Meisei University, Iwaki, Japan Email: *[email protected] Received October 13, 2013; revised November 12, 2013; accepted December 6, 2013 Copyright © 2013 Shuichi Miyamoto et al. This is an open access article distributed under the Creative Commons Attribution Li- cense, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. ABSTRACT Exodeoxyribonuclease III (EXOIII) acts as a 3’→5’ exonuclease and is homologous to purinic/apyrimidinic (AP) en- donuclease (APE), which plays an important role in the base excision repair pathway. To structurally investigate the reaction and substrate recognition mechanisms of EXOIII, a crystallographic study of EXOIII from Sulfolobus tokodaii strain 7 was carried out. The purified enzyme was crystallized by using the hanging-drop vapor-diffusion method. The crystals belonged to space group C2, with unit-cell parameters a = 154.2, b = 47.7, c = 92.4 Å, β = 125.8˚ and diffracted to 1.5 Å resolution. Keywords: Crenarchaeon; Crystallization; Exodeoxyribonuclease; Sulfolobus tokodaii; X-Ray Diffraction 1. Introduction formational change upon protein binding that permits complex formation and activation of attacking water, A variety of mechanisms exist to repair damaged DNA leading to incision, in the presence of Mg2+ [10,11].
    [Show full text]
  • Antiviral Activities of Oleanolic Acid and Its Analogues
    molecules Review Antiviral Activities of Oleanolic Acid and Its Analogues Vuyolwethu Khwaza, Opeoluwa O. Oyedeji and Blessing A. Aderibigbe * Department of Chemistry, University of Fort Hare, Alice Campus, Alice 5700, Eastern Cape, South Africa; [email protected] (V.K.); [email protected] (O.O.O) * Correspondence: [email protected]; Tel.: +27-406022266; Fax: +08-67301846 Academic Editors: Patrizia Ciminiello, Alfonso Mangoni, Marialuisa Menna and Orazio Taglialatela-Scafati Received: 27 July 2018; Accepted: 5 September 2018; Published: 9 September 2018 Abstract: Viral diseases, such as human immune deficiency virus (HIV), influenza, hepatitis, and herpes, are the leading causes of human death in the world. The shortage of effective vaccines or therapeutics for the prevention and treatment of the numerous viral infections, and the great increase in the number of new drug-resistant viruses, indicate that there is a great need for the development of novel and potent antiviral drugs. Natural products are one of the most valuable sources for drug discovery. Most natural triterpenoids, such as oleanolic acid (OA), possess notable antiviral activity. Therefore, it is important to validate how plant isolates, such as OA and its analogues, can improve and produce potent drugs for the treatment of viral disease. This article reports a review of the analogues of oleanolic acid and their selected pathogenic antiviral activities, which include HIV, the influenza virus, hepatitis B and C viruses, and herpes viruses. Keywords: HIV; influenza virus; HBV/HCV; natural product; triterpenoids; medicinal plant 1. Introduction Viral diseases remain a major problem for humankind. It has been reported in some reviews that there is an increase in the number of viral diseases responsible for death and morbidity around the world [1,2].
    [Show full text]
  • The Significance of N-Methylation of Bacillithiol on Its Biological Activity As a Redox Cofactor
    THE SIGNIFICANCE OF N-METHYLATION OF BACILLITHIOL ON ITS BIOLOGICAL ACTIVITY AS A REDOX COFACTOR Hazel Nicole Moxham A thesis submitted for the Degree of Master of Science by Research University of East Anglia School of Pharmacy September 2018 This copy of the thesis has been supplied on condition that anyone who consults it is understood to recognise that its copyright rests with the author and that use of any information derived therefrom must be in accordance with current UK Copyright Law. In addition, any quotation or extract must include full attribution. 1 Abstract Low molecular weight thiols play a crucial role in a multitude of biological processes such as maintaining redox homeostasis and the detoxification of chemical stressors. Different classes of microorganisms utilise different low molecular weight thiols. For example: glutathione is found eukaryotes and most gram-negative bacteria, mycothiol is found in the actinomycetes, and bacillithiol is found in the firmicutes. This study focused on N-methyl-bacillithiol, the novel low molecular weight thiol found in the green sulfur bacteria. Due to the unavailability of the thiol, the biophysical properties of a series of related derivatives were analysed and compared. Six thiols were examined so that each of their macroscopic and microscopic pKa values as well as their thiol-disulfide exchange rate constants and their copper catalysed autoxidation rates were isolated. The results determined that each thiol maintains its own set of biophysical properties that are unique to each compound. These were then observed alongside others within the literature to compare and contrast. Predictions were made regarding the properties of N-methylated bacillithiol by associating the data of those with similar structural differences.
    [Show full text]
  • 1Ako Lichtarge Lab 2006
    Pages 1–6 1ako Evolutionary trace report by report maker November 5, 2010 4.3.3 DSSP 5 4.3.4 HSSP 5 4.3.5 LaTex 5 4.3.6 Muscle 5 4.3.7 Pymol 5 4.4 Note about ET Viewer 6 4.5 Citing this work 6 4.6 About report maker 6 4.7 Attachments 6 1 INTRODUCTION From the original Protein Data Bank entry (PDB id 1ako): Title: Exonuclease iii from escherichia coli Compound: Mol id: 1; molecule: exonuclease iii; chain: a; ec: 3.1.11.2; engineered: yes Organism, scientific name: Escherichia Coli; 1ako contains a single unique chain 1akoA (268 residues long). 2 CHAIN 1AKOA 2.1 P09030 overview CONTENTS From SwissProt, id P09030, 100% identical to 1akoA: Description: Exodeoxyribonuclease III (EC 3.1.11.2) (Exonuclease 1 Introduction 1 III) (EXO III) (AP endonuclease VI). 2 Chain 1akoA 1 Organism, scientific name: Escherichia coli. 2.1 P09030 overview 1 Taxonomy: Bacteria; Proteobacteria; Gammaproteobacteria; 2.2 Multiple sequence alignment for 1akoA 1 Enterobacteriales; Enterobacteriaceae; Escherichia. 2.3 Residue ranking in 1akoA 1 Function: Major apurinic-apyrimidinic endonuclease of E.coli. It 2.4 Top ranking residues in 1akoA and their position on removes the damaged DNA at cytosines and guanines by cleaving the structure 2 on the 3’ side of the AP site by a beta-elimination reaction. It exhi- 2.4.1 Clustering of residues at 25% coverage. 2 bits 3’-5’-exonuclease, 3’-phosphomonoesterase, 3’-repair diesterase 2.4.2 Possible novel functional surfaces at 25% and ribonuclease H activities.
    [Show full text]
  • Plant-Derived Triterpenoid Biomarkers and Their Applications In
    Plant-derived triterpeonid biomarkers: chemotaxonomy, geological alteration, and vegetation reconstruction Res. Org. Geochem. 35, 11 − 35 (2019) Reviews-2015 Taguchi Award Plant-derived triterpenoid biomarkers and their applications in paleoenvironmental reconstructions: chemotaxonomy, geological alteration, and vegetation reconstruction Hideto Nakamura＊ （Received November 22, 2019; Accepted December 27, 2019） Abstract Triterpenoids and their derivatives are ubiquitous in sediment samples. Land plants are major sources of non- hopanoid triterpenoids; these terpenoids comprise a vast number of chemotaxonomically distinct biomolecules. Hence, geologically occurring plant-derived triterpenoids (geoterpenoids) potentially record unique characteristics of paleovegetation and sedimentary environments, and serve as source-specific markers for studying paleoenviron- ments. This review is aimed at explaining the origin of triterpenoids and their use as biomarkers in elucidating paleo- environments. Herein, application of plant-derived triterpenoids is discussed in terms of: (i) their biosynthetic pathways. These compounds are primarily synthesized via oxidosqualene cyclase (OSCs) and serve as precursors for a variety of membrane sterols and steroid hormones. Studies on OSCs and resulting compounds have helped elucidate the diversity and origin of the parent terpenoids. (ii) their chemotaxonomic significance. Geochemically important classes of triterpenoid skeletons are useful in gathering and substantiating information on botanical ori- gin of
    [Show full text]
  • Serine Proteases with Altered Sensitivity to Activity-Modulating
    (19) & (11) EP 2 045 321 A2 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: (51) Int Cl.: 08.04.2009 Bulletin 2009/15 C12N 9/00 (2006.01) C12N 15/00 (2006.01) C12Q 1/37 (2006.01) (21) Application number: 09150549.5 (22) Date of filing: 26.05.2006 (84) Designated Contracting States: • Haupts, Ulrich AT BE BG CH CY CZ DE DK EE ES FI FR GB GR 51519 Odenthal (DE) HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI • Coco, Wayne SK TR 50737 Köln (DE) •Tebbe, Jan (30) Priority: 27.05.2005 EP 05104543 50733 Köln (DE) • Votsmeier, Christian (62) Document number(s) of the earlier application(s) in 50259 Pulheim (DE) accordance with Art. 76 EPC: • Scheidig, Andreas 06763303.2 / 1 883 696 50823 Köln (DE) (71) Applicant: Direvo Biotech AG (74) Representative: von Kreisler Selting Werner 50829 Köln (DE) Patentanwälte P.O. Box 10 22 41 (72) Inventors: 50462 Köln (DE) • Koltermann, André 82057 Icking (DE) Remarks: • Kettling, Ulrich This application was filed on 14-01-2009 as a 81477 München (DE) divisional application to the application mentioned under INID code 62. (54) Serine proteases with altered sensitivity to activity-modulating substances (57) The present invention provides variants of ser- screening of the library in the presence of one or several ine proteases of the S1 class with altered sensitivity to activity-modulating substances, selection of variants with one or more activity-modulating substances. A method altered sensitivity to one or several activity-modulating for the generation of such proteases is disclosed, com- substances and isolation of those polynucleotide se- prising the provision of a protease library encoding poly- quences that encode for the selected variants.
    [Show full text]
  • Etats Rapides
    List of European Pharmacopoeia Reference Standards Effective from 2015/12/24 Order Reference Standard Batch n° Quantity Sale Information Monograph Leaflet Storage Price Code per vial Unit Y0001756 Exemestane for system suitability 1 10 mg 1 2766 Yes +5°C ± 3°C 79 ! Y0001561 Abacavir sulfate 1 20 mg 1 2589 Yes +5°C ± 3°C 79 ! Y0001552 Abacavir for peak identification 1 10 mg 1 2589 Yes +5°C ± 3°C 79 ! Y0001551 Abacavir for system suitability 1 10 mg 1 2589 Yes +5°C ± 3°C 79 ! Y0000055 Acamprosate calcium - reference spectrum 1 n/a 1 1585 79 ! Y0000116 Acamprosate impurity A 1 50 mg 1 3-aminopropane-1-sulphonic acid 1585 Yes +5°C ± 3°C 79 ! Y0000500 Acarbose 3 100 mg 1 See leaflet ; Batch 2 is valid until 31 August 2015 2089 Yes +5°C ± 3°C 79 ! Y0000354 Acarbose for identification 1 10 mg 1 2089 Yes +5°C ± 3°C 79 ! Y0000427 Acarbose for peak identification 3 20 mg 1 Batch 2 is valid until 31 January 2015 2089 Yes +5°C ± 3°C 79 ! A0040000 Acebutolol hydrochloride 1 50 mg 1 0871 Yes +5°C ± 3°C 79 ! Y0000359 Acebutolol impurity B 2 10 mg 1 -[3-acetyl-4-[(2RS)-2-hydroxy-3-[(1-methylethyl)amino] propoxy]phenyl] 0871 Yes +5°C ± 3°C 79 ! acetamide (diacetolol) Y0000127 Acebutolol impurity C 1 20 mg 1 N-(3-acetyl-4-hydroxyphenyl)butanamide 0871 Yes +5°C ± 3°C 79 ! Y0000128 Acebutolol impurity I 2 0.004 mg 1 N-[3-acetyl-4-[(2RS)-3-(ethylamino)-2-hydroxypropoxy]phenyl] 0871 Yes +5°C ± 3°C 79 ! butanamide Y0000056 Aceclofenac - reference spectrum 1 n/a 1 1281 79 ! Y0000085 Aceclofenac impurity F 2 15 mg 1 benzyl[[[2-[(2,6-dichlorophenyl)amino]phenyl]acetyl]oxy]acetate
    [Show full text]
  • Investigations Into Intracellular Thiols of Biological Importance
    Investigations into Intracellular Thiols of Biological Importance by Christine Elizabeth Hand A thesis presented to the University of Waterloo in fulfillment of the thesis requirement for the degree of Doctor of Philosophy in Chemistry Waterloo, Ontario, Canada, 2007 © Christine Elizabeth Hand 2007 AUTHOR'S DECLARATION I hereby declare that I am the sole author of this thesis. This is a true copy of the thesis, including any required final revisions, as accepted by my examiners. I understand that my thesis may be made electronically available to the public. ii Abstract The presence of thiols in living systems is critical for the maintenance of cellular redox homeostasis, the maintenance of protein thiol-disulfide ratios and the protection of cells from reactive oxygen species. In addition to the well studied tripeptide glutathione (γ-Glu-Cys-Gly), a number of compounds have been identified that contribute to these essential cellular roles. Many of these molecules are of great clinical interest due to their essential role in the biochemistry of a number of deadly pathogens, as well as their possible role as therapeutic agents in the treatment of a number of diseases. A series of studies were undertaken using theoretical, chemical and biochemical approaches on a selection of thiols, ergothioneine, the ovothiols and mycothiol, to further our understanding of these necessary biological components. Ergothioneine is present at significant physiological levels in humans and other mammals; however, a definitive role for this thiol has yet to be determined. It has been implicated in radical scavenging in vivo and shows promise as a therapeutic agent against disease states caused by oxidative damage.
    [Show full text]
  • Functional Identification of a Novel Gene, Moae, for 3
    www.nature.com/scientificreports OPEN Functional Identification of a Novel Gene, moaE, for 3-Succinoylpyridine Degradation in Received: 29 May 2015 Accepted: 28 July 2015 Pseudomonas putida S16 Published: 25 August 2015 Yi Jiang1,2, Hongzhi Tang1,2, Geng Wu1,2 & Ping Xu1,2 Microbial degradation of N-heterocyclic compounds, including xanthine, quinoline, nicotinate, and nicotine, frequently requires molybdenum hydroxylases. The intramolecular electron transfer chain of molybdenum hydroxylases consists of a molybdenum cofactor, two distinct [2Fe-2S] clusters, and flavin adenine dinucleotide. 3-Succinoylpyridine monooxygenase (Spm), responsible for the transformation from 3-succinoylpyridine to 6-hydroxy-3-succinoylpyridine, is a crucial enzyme in the pyrrolidine pathway of nicotine degradation in Pseudomonas. Our previous work revealed that the heterotrimeric enzyme (SpmA, SpmB, and SpmC) requires molybdopterin cytosine dinucleotide as a cofactor for their activities. In this study, we knocked out four genes, including PPS_1556, PPS_2936, PPS_4063, and PPS_4397, and found that a novel gene, PPS_4397 encoding moaE, is necessary for molybdopterin cytosine dinucleotide biosynthesis. Resting cell reactions of the moaE deletion mutant incubated with 3 g l−1 nicotine at 30 °C resulted in accumulation of 3-succinoylpyridine, and the strain complemented by the moaE gene regained the ability to convert 3-succinoylpyridine. In addition, reverse transcription-quantitative polymerase chain reaction analysis indicated that the transcriptional levels of the genes of moaE, spmA, and spmC of Pseudomonas putida S16 were distinctly higher when grown in nicotine medium than in glycerol medium. Large quantities of tobacco wastes containing high concentration of nicotine are produced during tobacco manufacturing yearly1. Nicotine is well known to be harmful to human health and can cross biological membranes and blood-brain barrier easily2.
    [Show full text]
  • (12) United States Patent (10) Patent No.: US 7,794,716 B2 Adair (45) Date of Patent: *Sep
    US007794,716 B2 (12) United States Patent (10) Patent No.: US 7,794,716 B2 Adair (45) Date of Patent: *Sep. 14, 2010 (54) ANTIBODY COMPOSITION AND PASSIVE Adair, C.D. et al. “Elevated Endoxin-Like Factor Complicating a MMUNIZATION AGAINST Multifetal Second Trimester Pregnancy: Treatment Digoxin-Binding PREGNANCY-INDUCED HYPERTENSION Immunoglobulin'. Am. J. Nephrol., 1996, vol. 16, pp. 529-531. Aizman, O. et al., “Ouabain, a steroid hormone that signals with slow (75) Inventor: Charles David Adair, Signal Mountain, calcium oscillations'. PNAS, 2001, vol.98, No. 23, pp. 13420-13424. TN (US) Amorium, M.M.R., et al., “Corticosteriod therapy for prevention of respiratory distress syndrome in severe preeclampsia'. Am. J. Obstet. (73) Assignee: Glenveigh Pharmaceuticals, LLC, Gyngol., 1999, vol. 180, No. 5, pp. 1283-1288. Chattanooga, TN (US) Bagrov, A. Y., et al., “Characterizatin of a Urinary Bufodienolide Na+, K+-ATPase Inhibitor in Patients. After Acute Myocardial Infarc (*) Notice: Subject to any disclaimer, the term of this tion'. Hypertension, 1998, vol. 31, pp. 1097-1 103. patent is extended or adjusted under 35 Ball, W. J. Jr. et al., “Isolation and Characterization of Human Monoclonal Antibodies to Digoxin'. The Journal of Immunology, U.S.C. 154(b) by 700 days. 1999, vol. 163, pp. 2291-2298. This patent is Subject to a terminal dis Butler, V. et al., “Digoxin-Specific Antibodies'. Proc. Natl. Acad. Sci. claimer. USA (Physiology), 1967, vol. 57, pp. 71-78. Dasgupta, A. et al., “Monitoring Free Digoxin Instead of Total Digoxin in Patients with Congestive Heart Failure and High Concen (21) Appl. No.: 11/317,378 trations of Dogoxin-like Immunoreactive Substances”.
    [Show full text]