DOCSLIB.ORG

Complete Thesis

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Complete Thesis University of Groningen Nonribosomal peptide synthetases Zwahlen, Reto Daniel IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from it. Please check the document version below. Document Version Publisher's PDF, also known as Version of record Publication date: 2018 Link to publication in University of Groningen/UMCG research database Citation for published version (APA): Zwahlen, R. D. (2018). Nonribosomal peptide synthetases: Engineering, characterization and biotechnological potential. University of Groningen. Copyright Other than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons). The publication may also be distributed here under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license. More information can be found on the University of Groningen website: https://www.rug.nl/library/open-access/self-archiving-pure/taverne- amendment. Take-down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons the number of authors shown on this cover page is limited to 10 maximum. Download date: 04-10-2021 Nonribosomal peptide synthetases: Engineering, characterization and biotechnological potential Academic Thesis, University of Groningen, the Netherlands ISBN: 978-94-034-0674-9 978-94-034-0673-2 (e-book) Printing: Eikon + Cover: Reto D. Zwahlen & Lovebird design. Layout: Lovebird design. www.lovebird-design.com © R. D. Zwahlen, Groningen, the Netherlands, 2018 All rights reserved. No part of this publication may be reproduced or transmitted in any form or by any means, without written permission of the author. Nonribosomal peptide synthetases: Engineering, characterization and biotechnological potential PhD thesis to obtain the degree of PhD at the University of Groningen on the authority of the Rector Magnificus Prof. E. Sterken and in accordance with the decision by the College of Deans. This thesis will be defended in public on The 18th of May 2018 at 14:30 hours by Reto Daniel Zwahlen born on 24 July 1988 in Bern, Switzerland Supervisors Prof. A.J.M. Driessen Prof. R.A.L. Bovenberg Assessment committee Prof. D.B. Janssen Prof. J. Raaijmakers Prof. L. Dijkhuizen For you, my love, my other half, my Tonia. Table of contents Chapter I Introduction 9 Chapter II Identification and characterization of nonribosomal peptide synthe- tase modules that activate 4-hydroxyphenylglycine 45 Chapter III A golden gate based system for convenient assembly of chimeric Nonribosomal peptide synthetases 71 Chapter IV Biochemical and structural characterization of the Nocardia lactamdu- rans L-δ-(α-aminoadipyl)-L-cysteinyl-D-valine synthetase 89 Chapter V An engineered two component nonribosomal peptide synthetase (NRPS) producing a novel peptide-like compound in Penicillium chrysogenum 117 Chapter VI Prokaryotic MbtH like proteins stimulate secondary metabolism in the filamentous fungus Penicillium chrysogenum 145 Chapter VII Summary and outlook 179 Deutsche Zusammenfassung 187 Nederlandse samenvatting 197 Appendices Acknowledgements 205 List of publications and patents 209 CHAPTER I Introduction Reto D. Zwahlen, Roel A.L. Bovenberg,2,3 and Arnold J.M. Driessen1,4 1Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands 2Synthetic Biology and Cell Engineering, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands 3DSM Biotechnology Centre, Delft, The Netherlands 4Kluyver Centre for Genomics of Industrial Fermentations, Julianalaan 67, 2628BC Delft, The Netherlands In part to be published as a review in Frontiers in Microbiology, abstract accepted Abstract Secondary metabolites represent a class of bioactive natural prod- ucts and have a long history for human application and are a cor- nerstone of medicine and chemistry where they have found wide- spread recognition. Modern medicine has among other factors been enabled through the discovery and therapeutical implemen- tation of secondary metabolites, such as antibiotics or anti-tumor agents and up until this day are a vital component to contempo- rary medicine. The origin of these products can be traced to plants, fungi and bacteria alike. The underlying biosynthetic clusters fre- quently contain a multiverse of distinct genes encoding a series of enzymes who interact in a specific manner to ultimately produce a particular product. At the very core of such a cluster is predomi- nantly a complex and multi-modular molecular machinery or more precisely a nonribosomal peptide synthetase (NRPS). NRPS along with related enzymes represent an outstanding class of secondary metabolite producers which have been a focus point in the dis- covery and engineering of novel bioactive compounds for decades. This chapter will give an introduction to natural products, the asso- ciated enzymes and the evolution of accompanying tools for their discovery and engineering with a focus on NRPS. 1. Nonribosomal peptide synthetases (NRPS) and nonribosomal peptides (NRP) 1 Nonribosonal peptide synthetases (NRPS) are large, highly structured and complex enzymatic machineries, closely related to other modular enzymes such as polyketide synthetases (PKS), NRPS-PKS hybrid synthetase and fatty acid synthetases (FAS). They have certain distinct properties in common, the Introduction most striking one being their structural division in domains and modules, which is manifested in their shared evolutionary history (Smith and Sherman 2008). Every enzyme minimally consists of one module, a functionally dis- tinct unit, which allows for the recruitment and subsequent incorporation of a precursor into a growing product. Every NRPS module, initiation (1), elon- gation (n) or termination (1), requires a minimal set of domains [2]. The two domains essential to every module are the adenylation domain (A) and the non-catalytic thiolation domain (T). This tandem di-domain enables the spe- cific selection and activation of a given substrate. However, the T-domain must first go through 4’-phosphopantetheinyl transferase (PPtase) and co- enzyme A (CoA) dependent activation after expression, by transferring a phoshpantethetheine moiety to a conserved serine residue, in order to enter an active state. Also, adenylation domains (A) have accompanying factors, or proteins, called MbtH-like proteins (MLP) [3–4]. In contrast to PPtases, MLPs are merely interacting with the A-domain, however they do not have an intrinsic enzymatic activity, though rather a chaperoning function upon binding a distinct part of the A domain [5–7]. In addition to these domains, any elongation module will require a condensation domain (C), which con- nects two modules and links up- and downstream activated substrates via a peptide bond. C-domains are stereospecific for both, up- and downstream activated substrates and transfer the resulting intermediate compound to the downstream T-domain. Lastly, the C-terminal termination module es- sentially requires a thioesterase domain (Te), to catalytically release the co- valently bound compound of the NRPS, returning the NRPS complex to the ground state for another reaction cycle. In addition to these essential do- mains, we can distinguish a series of additional domains, performing epi- merization, halogenation, cyclization, macrocyclization, multimerization or methylation [8–10]. Domains as well as modules are clearly defined and evo- lutionary exchangeable structures amongst multi-modular enzymes. In the case of PKS and NRPS, this lead to the occurrence of a variety of NRPS-PKS hybrids [11–14]. A NRPS can be as simple as a single modular unit containing three domains, although the most complex and largest structure known con- tains 15 modules with 46 domains [15–16] yielding a 1.8 mD protein complex 11 (type I NRPS). Although the size of a NRPS, as well as the modular sequence, limits the size of an NRPS product, it is common that NRPS enzymes cluster and interact with tailoring enzymes in order to produce products of a higher complexity [17]. To enable such specific interactions, NRPS can contain small stretches of up to 30 amino acids at the C- or N-terminus, which form a rather specific recognition point, thus enabling communication (COM-domain) be- tween multiple NRPS of one cluster (type II NRPS) [18–19]. As a result of this multitude of functional dimensions within the NRPS architecture, and the increasing number of NRPS containing biosynthetic gene clusters there is a large growing reservoir of natural products with potentially new properties. NRPS are a crucial part of the secondary metabolism, which is restricted to different prokaryotic, fungal and certain higher eukaryotic species [20]. In comparison to most ribosomally derived peptides, nonribosomal pep- tides (NRPs) are low molecular weight products. The structural diversity of NRPs is tremendous, mostly due to their chemical complexity. Significantly contributing to this diversity is the fact that NRPS are not only reliant on proteinogenic amino acids only, but up until now more than 500 substrates were
Load more

Recommended publications
  • Genes in Eyecare Geneseyedoc 3 W.M
    Genes in Eyecare geneseyedoc 3 W.M. Lyle and T.D. Williams 15 Mar 04 This information has been gathered from several sources; however, the principal source is V. A. McKusick’s Mendelian Inheritance in Man on CD-ROM. Baltimore, Johns Hopkins University Press, 1998. Other sources include McKusick’s, Mendelian Inheritance in Man. Catalogs of Human Genes and Genetic Disorders. Baltimore. Johns Hopkins University Press 1998 (12th edition). http://www.ncbi.nlm.nih.gov/Omim See also S.P.Daiger, L.S. Sullivan, and B.J.F. Rossiter Ret Net http://www.sph.uth.tmc.edu/Retnet disease.htm/. Also E.I. Traboulsi’s, Genetic Diseases of the Eye, New York, Oxford University Press, 1998. And Genetics in Primary Eyecare and Clinical Medicine by M.R. Seashore and R.S.Wappner, Appleton and Lange 1996. M. Ridley’s book Genome published in 2000 by Perennial provides additional information. Ridley estimates that we have 60,000 to 80,000 genes. See also R.M. Henig’s book The Monk in the Garden: The Lost and Found Genius of Gregor Mendel, published by Houghton Mifflin in 2001 which tells about the Father of Genetics. The 3rd edition of F. H. Roy’s book Ocular Syndromes and Systemic Diseases published by Lippincott Williams & Wilkins in 2002 facilitates differential diagnosis. Additional information is provided in D. Pavan-Langston’s Manual of Ocular Diagnosis and Therapy (5th edition) published by Lippincott Williams & Wilkins in 2002. M.A. Foote wrote Basic Human Genetics for Medical Writers in the AMWA Journal 2002;17:7-17. A compilation such as this might suggest that one gene = one disease.
    [Show full text]
  • Teleomorph Hypocrea Jecorina)
    Downloaded from orbit.dtu.dk on: Dec 20, 2017 Unraveling the Secondary Metabolism of the Biotechnological Important Filamentous Fungus Trichoderma reesei ( Teleomorph Hypocrea jecorina) Jørgensen, Mikael Skaanning; Larsen, Thomas Ostenfeld; Mortensen, Uffe Hasbro; Aubert, Dominique Publication date: 2013 Document Version Publisher's PDF, also known as Version of record Link back to DTU Orbit Citation (APA): Jørgensen, M. S., Larsen, T. O., Mortensen, U. H., & Aubert, D. (2013). Unraveling the Secondary Metabolism of the Biotechnological Important Filamentous Fungus Trichoderma reesei ( Teleomorph Hypocrea jecorina). Kgs. Lyngby: Technical University of Denmark (DTU). General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Unraveling the Secondary Metabolism of the Biotechnological Important Filamentous Fungus Trichoderma reesei (Teleomorph Hypocrea jecorina) MJ-T-030 Mikael Skaanning Jørgensen Ph.D. Thesis November 2012 MJ-T-020 MJ-T-032 28 8 20 27 5 28 1 30 17 8 24 25 9 25 31 13 15 26 16 4 24 12 32 18 11 3 14 13 19 10 5 7 2 9 2521 22 23 Unraveling the Secondary Metabolism of the Biotechnological Important Filamentous Fungus Trichoderma reesei (Teleomorph Hypocrea jecorina) Mikael Skaanning Jørgensen Ph.D.
    [Show full text]
  • The Inducers 1,3-Diaminopropane and Spermidine Cause The
    JOURNAL OF PROTEOMICS 85 (2013) 129– 159 Available online at www.sciencedirect.com www.elsevier.com/locate/jprot The inducers 1,3-diaminopropane and spermidine cause the reprogramming of metabolism in Penicillium chrysogenum, leading to multiple vesicles and penicillin overproduction Carlos García-Estradaa,⁎, Carlos Barreiroa, Mohammad-Saeid Jamia, Jorge Martín-Gonzáleza, Juan-Francisco Martínb,⁎⁎ aINBIOTEC, Instituto de Biotecnología de León, Avda. Real no. 1, Parque Científico de León, 24006 León, Spain bÁrea de Microbiología, Departamento de Biología Molecular, Universidad de León, Campus de Vegazana s/n; 24071 León, Spain ARTICLE INFO ABSTRACT Article history: In this article we studied the differential protein abundance of Penicillium chrysogenum in Received 21 February 2013 response to either 1,3-diaminopropane (1,3-DAP) or spermidine, which behave as inducers Accepted 15 April 2013 of the penicillin production process. Proteins were resolved in 2-DE gels and identified by Available online 30 April 2013 tandem MS spectrometry. Both inducers produced largely identical changes in the proteome, suggesting that they may be interconverted and act by the same mechanism. Keywords: The addition of either 1,3-DAP or spermidine led to the overrepresentation of the last 1,3-diaminopropane enzyme of the penicillin pathway, isopenicillin N acyltransferase (IAT). A modified form of Spermidine the IAT protein was newly detected in the polyamine-supplemented cultures. Both Penicillin inducers produced a rearrangement of the proteome resulting in an overrepresentation of Penicillium chrysogenum enzymes involved in the biosynthesis of valine and other precursors (e.g. coenzyme A) of Vesicles penicillin. Interestingly, two enzymes of the homogentisate pathway involved in the degradation of phenylacetic acid (a well-known precursor of benzylpenicillin) were reduced following the addition of either of these two inducers, allowing an increase of the phenylacetic acid availability.
    [Show full text]
  • Structural Basis for the Activation of Phenylalanine in the Non-Ribosomal Biosynthesis of Gramicidin S
    The EMBO Journal Vol.16 No.14 pp.4174–4183, 1997 Structural basis for the activation of phenylalanine in the non-ribosomal biosynthesis of gramicidin S Elena Conti1,2, Torsten Stachelhaus3, the peptide product. Each amino acid is activated by Mohamed A.Marahiel3 and Peter Brick1,4 adenylation of its carboxylate group with ATP and then transferred to the thiol group of an enzyme-bound phospho- 1Biophysics Section, Blackett Laboratory, Imperial College, pantetheine cofactor for possible modification and the 3 London SW7 2BZ, UK and Biochemie/Fachbereich Chemie, elongation reaction (Stachelhaus and Marahiel, 1995a; Philipps-Universita¨t Marburg, D-35032 Marburg, Germany Kleinkauf and von Do¨hren, 1996). 2 Present address: Laboratory of Molecular Biophysics, The cloning and sequencing of several peptide synthe- Rockefeller University, New York, NY10021, USA tase genes have revealed a conserved and ordered modular 4Corresponding author organization. Each module encodes a functional building unit containing ~1000 amino acids, which specifically The non-ribosomal synthesis of the cyclic peptide recognizes a single amino acid. Within such a protein antibiotic gramicidin S is accomplished by two large template-directed peptide biosynthesis, the occurrence and multifunctional enzymes, the peptide synthetases 1 and specific order of the modules in the genomic DNA dictate 2. The enzyme complex contains five conserved subunits the number and sequence of the amino acids to be of ~60 kDa which carry out ATP-dependent activation incorporated into the resulting oligopeptide. The modular of specific amino acids and share extensive regions of arrangement of peptide synthetases closely parallels the sequence similarity with adenylating enzymes such multienzyme complexes responsible for the biogenesis of as firefly luciferases and acyl-CoA ligases.
    [Show full text]
  • Three Conserved Glycine Residues in Valine Activation of Gramicidin S Synthetase 2 from Bacillus Brevis1
    J. Biochem. 117, 276-282 (1995) Three Conserved Glycine Residues in Valine Activation of Gramicidin S Synthetase 2 from Bacillus brevis1 Masaki Saito, Kazuko Hori, Toshitsugu Kurotsu, Masayuki Kanda, and Yoshitaka Saito Department of Biochemistry, Hyogo College of Medicine, Mukogawa-cho, Nishinomiya, Hyogo 663 Received for publication, July 27, 1994 The translated product from the gene fragment containing the second and third domains of gramicidin S synthetase 2 was purified to an essentially homogeneous state. It showed valine and ornithine-activating activity and the second domain was proved to be the valine-activating domain. Three mutant genes from Bacillus brevis Nagano, BI-3, E-4, and E-5 strains, which encode defective valine-activating domains of gramicidin S synthetase 2, were sequenced. By comparison with the wild-type gene, single point mutations of guanine to adenine were found at the three conserved glycine codons; the 5303rd guanine in BI-3, the 5378th guanine in E-4, and the 4967th guanine in E-5, which corresponded to codon changes of the 1768th glycine to glutamic acid and the 1793rd and the 1656th glycine to aspartic acid. Loss of valine-adenylation activity by mutation at the 1656th glycine proved the direct participation of the TSGT/STGXPKG motif in the adenylation reaction, and suggests that this glycine residue with the conserved lysine residue of the motif forms the phosphate-binding loop for ATP-binding. The 1793rd glycine is a member of the YGXTE motif which was also conserved among adenylate-forming enzymes except acetyl-CoA synthetases. The 1768th glycine residue appears to maintain the conformation of the active site for aminoacyl adenylation since this residue is retained among the adenylate-forming enzymes, though flanking regions are not conserved.
    [Show full text]
  • Omics Approaches Applied to Penicillium Chrysogenum and Penicillin Production: Revealing the Secrets of Improved Productivity
    G C A T T A C G G C A T genes Review Omics Approaches Applied to Penicillium chrysogenum and Penicillin Production: Revealing the Secrets of Improved Productivity Carlos García-Estrada 1,2,*, Juan F. Martín 3, Laura Cueto 1 and Carlos Barreiro 1,4 1 INBIOTEC (Instituto de Biotecnología de León). Avda. Real 1—Parque Científico de León, 24006 León, Spain; [email protected] (L.C.); [email protected] (C.B.) 2 Departamento de Ciencias Biomédicas, Universidad de León, Campus de Vegazana s/n, 24071 León, Spain 3 Área de Microbiología, Departamento de Biología Molecular, Facultad de Ciencias Biológicas y Ambientales, Universidad de León, 24071 León, Spain; [email protected] 4 Departamento de Biología Molecular, Universidad de León, Campus de Ponferrada, Avda. Astorga s/n, 24401 Ponferrada, Spain * Correspondence: [email protected] or [email protected]; Tel.: +34-987210308 Received: 22 April 2020; Accepted: 24 June 2020; Published: 26 June 2020 Abstract: Penicillin biosynthesis by Penicillium chrysogenum is one of the best-characterized biological processes from the genetic, molecular, biochemical, and subcellular points of view. Several omics studies have been carried out in this filamentous fungus during the last decade, which have contributed to gathering a deep knowledge about the molecular mechanisms underlying improved productivity in industrial strains. The information provided by these studies is extremely useful for enhancing the production of penicillin or other bioactive secondary metabolites by means of Biotechnology or Synthetic Biology. Keywords: penicillin; Penicillium chrysogenum; omics; beta-lactam antibiotics 1. Introduction There are few examples of industrial microbial processes as deeply characterized as penicillin production by the filamentous fungus Penicillium chrysogenum.
    [Show full text]
  • Structural Studies of Natural Product Biosynthetic Proteins Craig a Townsend
    Review 721 Structural studies of natural product biosynthetic proteins Craig A Townsend The first high-resolution structures of key proteins involved in the Introduction biosynthesis of several natural product classes are now appear- X-ray qstallography has dramatically advanced our under- ing. In some cases, they have resulted in a significantly improved standing of protein structure and the macromolecular asso- mechanistic understanding of the often complex processes ciation of proteins with IlKA, RNA, drugs, cofactors and catalyzed by these enzymes, and they have also opened the way other proteins. Progress has, however, been considerably for more rational efforts to modify the products made. slower in obtaining detailed insights into the enzymes that catalyze natural product biosynthesis. ‘I’he reasons for this Address: Department of Chemistry, The Johns Hopkins University, Baltimore, MD 21218, USA. are several; in particular the enzymes concerned have modest, even poor, kinetic properties, they are available in Correspondence: Craig A Townsend only minute amounts from wild-type organisms and they E-mail: [email protected] are often not very stable. As a consequence, they have proved difficult to detect, isolate and purify. Efforts towards Chemistry & Biology October 1997, 4:721-730 http://biomednet.com/elecref/1074552100400721 this end have been successful in the last decade, ho\vever, and now a number of genes encoding synthetic enzymes for C Current Biology Ltd ISSN 1074-5521 natural products of interest have been cloned and recombi- nant enzymes have been made available for thorough char- acterization. Given the commercial importance of natural products, the associated enzymes hai,e become targets for manipulation by chemical modification, muragenesis and combinatorial methods, not onI\; to improve the production of these high-value materials but also, through deeper mechanistic understanding, to achieve the efficient synthe- sis of new chemical entities.
    [Show full text]
  • University of Groningen Genome Sequencing and Analysis Of
    University of Groningen Genome sequencing and analysis of the filamentous fungus Penicillium chrysogenum van den Berg, Marco A.; Albang, Richard; Albermann, Kaj; Badger, Jonathan H.; Daran, Jean-Marc; Driessen, Arnold; Garcia-Estrada, Carlos; Fedorova, Natalie D.; Harris, Diana M.; Heijne, Wilbert H. M. Published in: Nature Biotechnology DOI: 10.1038/nbt.1498 IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from it. Please check the document version below. Document Version Publisher's PDF, also known as Version of record Publication date: 2008 Link to publication in University of Groningen/UMCG research database Citation for published version (APA): van den Berg, M. A., Albang, R., Albermann, K., Badger, J. H., Daran, J-M., Driessen, A. J. M., ... Bovenberg, R. A. L. (2008). Genome sequencing and analysis of the filamentous fungus Penicillium chrysogenum. Nature Biotechnology, 26(10), 1161-1168. DOI: 10.1038/nbt.1498 Copyright Other than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons). Take-down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons the number of authors shown on this cover page is limited to 10 maximum.
    [Show full text]
  • Saccharomyces Cerevisiae As Host for the Recombinant Production of Polyketides and Nonribosomal Peptides Anna Tippelt and Markus Nett*
    Tippelt and Nett Microb Cell Fact (2021) 20:161 https://doi.org/10.1186/s12934-021-01650-y Microbial Cell Factories REVIEW Open Access Saccharomyces cerevisiae as host for the recombinant production of polyketides and nonribosomal peptides Anna Tippelt and Markus Nett* Abstract As a robust, fast growing and genetically tractable organism, the budding yeast Saccharomyces cerevisiae is one of the most widely used hosts in biotechnology. Its applications range from the manufacturing of vaccines and hor- mones to bulk chemicals and biofuels. In recent years, major eforts have been undertaken to expand this portfolio to include structurally complex natural products, such as polyketides and nonribosomally synthesized peptides. These compounds often have useful pharmacological properties, which make them valuable drugs for the treatment of infectious diseases, cancer, or autoimmune disorders. In nature, polyketides and nonribosomal peptides are gener- ated by consecutive condensation reactions of short chain acyl-CoAs or amino acids, respectively, with the substrates and reaction intermediates being bound to large, multidomain enzymes. For the reconstitution of these multistep catalytic processes, the enzymatic assembly lines need to be functionally expressed and the required substrates must be supplied in reasonable quantities. Furthermore, the production hosts need to be protected from the toxicity of the biosynthetic products. In this review, we will summarize and evaluate the status quo regarding the heterologous production of polyketides
    [Show full text]
  • EUROPEAN PATENT OFFICE, VIENNA Thousand Oaks, CA 91320 (US) SUB-OFFICE
    Europäisches Patentamt *EP001033405A2* (19) European Patent Office Office européen des brevets (11) EP 1 033 405 A2 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: (51) Int Cl.7: C12N 15/29, C12N 15/82, 06.09.2000 Bulletin 2000/36 C07K 14/415, C12Q 1/68, A01H 5/00 (21) Application number: 00301439.6 (22) Date of filing: 25.02.2000 (84) Designated Contracting States: • Brover, Vyacheslav AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU Calabasas, CA 91302 (US) MC NL PT SE • Chen, Xianfeng Designated Extension States: Los Angeles, CA 90025 (US) AL LT LV MK RO SI • Subramanian, Gopalakrishnan Moorpark, CA 93021 (US) (30) Priority: 25.02.1999 US 121825 P • Troukhan, Maxim E. 27.07.1999 US 145918 P South Pasadena, CA 91030 (US) 28.07.1999 US 145951 P • Zheng, Liansheng 02.08.1999 US 146388 P Creve Coeur, MO 63141 (US) 02.08.1999 US 146389 P • Dumas, J. 02.08.1999 US 146386 P , (US) 03.08.1999 US 147038 P 04.08.1999 US 147302 P (74) Representative: 04.08.1999 US 147204 P Bannerman, David Gardner et al More priorities on the following pages Withers & Rogers, Goldings House, (83) Declaration under Rule 28(4) EPC (expert 2 Hays Lane solution) London SE1 2HW (GB) (71) Applicant: Ceres Incorporated Remarks: Malibu, CA 90265 (US) THE COMPLETE DOCUMENT INCLUDING REFERENCE TABLES AND THE SEQUENCE (72) Inventors: LISTING IS AVAILABLE ON CD-ROM FROM THE • Alexandrov, Nickolai EUROPEAN PATENT OFFICE, VIENNA Thousand Oaks, CA 91320 (US) SUB-OFFICE.
    [Show full text]
  • A-Aminoadipyl-Cysteinyl-Valine Synthetases in J8-Lactam Producing Organisms
    VOL. 53 NO. 10, OCT.2000 THE JOURNAL OF ANTIBIOTICS pp.1008 - 1021 REVIEW ARTICLE a-Aminoadipyl-cysteinyl-valine Synthetases in j8-Lactam Producing Organisms From Abraham's Discoveries to Novel Concepts of Non-Ribosomal Peptide Synthesis Juan F. Martin Area of Microbiology, Faculty of Biology, University of Leon, 2407 1 Leon, and Institute of Biotechnology, INBIOTEC, Science Park of Leon, Avda. del Real, n° 1, 24006 Leon, Spain (Received for publication July 27, 2000) The tripeptide 5-(L-a-aminoadipyl)-L-cysteinyl-D-valine (ACV) was discovered by Arnstein and Morris in Penicillium chrysogenum and Abraham and coworkers in Acremoniumchrysogenum.Other analogous tripeptides and tetrapeptides werelater reported in these and other /3-lactam producing fungi and actinomycetes. The ACVtripeptide is synthesized by a large non-ribosomal peptide synthetase named ACVsynthetase encoded by the 1 1 kb pcbAB gene. This gene has been cloned from the DNAof four different filamentous fungi and two actinomycetes. Detailed analysis of the multifunctional ACVsynthetases reveals that they consist of three repeated modules (initially named domains) involved in activation of the corresponding amino acids L-a-aminoadipic acid, L-cysteine and L-valine. Each module consists of functional domains for amino acid activation (A), condensation (C) and thiolation (T). In addition the last module of the ACVsynthetase contains an epimerization domain (E) involved in conversion of the L-valine to its D-isomer when the tripeptide is still enzyme linked. There are seven epimerization motifs conserved in the third module of all ACVsynthetases. In addition, there is an integrated thioesterase domain in the C-terminal region of the ACV synthetases that appears to be involved in the selective release of the tripeptide with the correct lld configuration.
    [Show full text]
  • ELISA Kit Catalog
    CATALOG & PRICE ELISA Kits 2020 Advanced BioChemicals, LLC www.advancedbiochemicals.com PO Box 465066 [email protected] Lawrenceville, GA 30042 Phone: 1-470-292-8602 USA 2020 Fax: 1-509-696-4035 Contents v Order Information v Additional Information v Human ELISA Kits (Page 1 – 30) v Mouse ELISA Kits (Page 31 – 44) v Rat ELISA Kits (Page 45 – 57) v Porcine ELISA Kits (Page 58 – 82) v Monkey ELISA Kits (Page 83 – 112) v Others: [email protected] Advanced BioChemicals, LLC www.advancedbiochemicals.com PO Box 465066 [email protected] Lawrenceville, GA 30042 Phone: 1-470-292-8602 USA 2020 Fax: 1-509-696-4035 Order Information To place an order with credit card: • Please use our secure online order system or provide credit card information by Fax: (+1)509-696-4035; Email: [email protected]; Tel: (+1) 470-292-8602. To place an order with a Purchase Order (PO) number: • Fax your PO to (+1) 509-696-4035; • Email your PO to [email protected]; • Call (+1) 470-292-8602 with your PO number • Mail your order form to: Advanced BioChemicals, LLC PO Box 465066, Lawrenceville, GA 30042, USA Additional Information • Visit our website www.advancedbiochemicals.com to get the latest product information and promotions; • Alternative package sizes available, contact us for more discount; • Terms and conditions applied to all orders and services. Terms and conditions can be found at the https://advancedbiochemicals.com/terms-of-use/. Advanced BioChemicals, LLC www.advancedbiochemicals.com PO Box 465066 [email protected] Lawrenceville, GA 30042 Phone: 1-470-292-8602 USA 2020 Fax: 1-509-696-4035 ELISA Kits Catalog - Human Advanced BioChemicals (ABC) www.advancedbiochemicals.com ABC Cat.
    [Show full text]