DOCSLIB.ORG

Biosynthesis and Engineering of Cyclomarin and Cyclomarazine: Prenylated, Non-Ribosomal Cyclic Peptides of Marine Actinobacterial Origin

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Biosynthesis and Engineering of Cyclomarin and Cyclomarazine: Prenylated, Non-Ribosomal Cyclic Peptides of Marine Actinobacterial Origin UC San Diego Research Theses and Dissertations Title Biosynthesis and Engineering of Cyclomarin and Cyclomarazine: Prenylated, Non-Ribosomal Cyclic Peptides of Marine Actinobacterial Origin Permalink https://escholarship.org/uc/item/21b965z8 Author Schultz, Andrew W. Publication Date 2010 Peer reviewed eScholarship.org Powered by the California Digital Library University of California UNIVERSITY OF CALIFORNIA, SAN DIEGO Biosynthesis and Engineering of Cyclomarin and Cyclomarazine: Prenylated, Non-Ribosomal Cyclic Peptides of Marine Actinobacterial Origin A dissertation submitted in partial satisfaction of the requirements for the degree Doctor of Philosophy in Oceanography by Andrew William Schultz Committee in charge: Professor Bradley Moore, Chair Professor Eric Allen Professor Pieter Dorrestein Professor William Fenical Professor William Gerwick 2010 Copyright Andrew William Schultz, 2010 All rights reserved. The Dissertation of Andrew William Schultz is approved, and it is acceptable in quality and form for publication on microfilm and electronically: ________________________________________________________________ ________________________________________________________________ ________________________________________________________________ Chair University of California, San Diego 2010 iii DEDICATION To my wife Elizabeth and our son Orion and To my parents Dale and Mary Thank you for your never ending love and support iv TABLE OF CONTENTS Signature Page ..................................................................................................... iii Dedication ............................................................................................................ iv Table of Contents ................................................................................................. v List of Abbreviations ............................................................................................. ix List of Figures ...................................................................................................... xii List of Tables ..................................................................................................... xvii Acknowledgements .......................................................................................... xviii Vita ..................................................................................................................... xxi Abstract of the Dissertation ............................................................................... xxii Chapter 1: Introduction: The Chemical Diversity of Non-ribosomal Peptides .............................................................................................................. 1 1.1: Introduction to non-ribosomal peptides .............................................. 2 1.2: NRPs are assembled via non-ribosomal peptide synthetases ......... 12 1.3: Tailoring….. ..................................................................................... 16 1.3.1: Epimerization ..................................................................... 16 1.3.2: Oxidation ............................................................................ 16 1.3.3: Halogenation ...................................................................... 21 1.3.4: Methylation ......................................................................... 22 1.3.5: Prenylation ......................................................................... 23 1.3.6: Glycosolation ...................................................................... 24 1.3.7: Cyclization of serine, threonine, or cysteine to form oxazoline or thiazoline ........................................................ 24 1.3.8: Reductive offloading ........................................................... 25 1.4: Biosynthetic pathways to non-proteinogenic amino acids ................ 26 v 1.4.1: Direct intermediates of primary metabolic pathways .......... 28 1.4.2: Transaminaition of α-keto acids ......................................... 28 1.4.3: Polyketide synthase derived ............................................... 29 1.4.5: Shikimate pathway ............................................................. 30 1.4.6: Aldolase/dehydrogenase heterodimers .............................. 33 1.5: Interfacing non-ribosomal peptide synthetases with polyketide synthases adds further chemical diversity ......................................... 33 1.6: Engineering of nonribosomal peptides to increase nature’s peptide diversity ............................................................................................. 34 1.6.1: Precursor directed biosynthesis and mutasynthesis .......... 35 1.6.2: Combinatorial biosynthesis ................................................ 40 1.7: Conclusion ....................................................................................... 43 1.8: References ...................................................................................... 46 Chapter 2: Biosynthesis of Cyclomarins and Cyclomarazines, Prenylated Cyclic Peptides of Marine Actinobacterial Origin .......................................... 55 2.1: Abstract....... ...................................................................................... 56 2.2 Introduction... ..................................................................................... 57 2.2.1: Cyclomarin consists of several modified and non- proteinogenic amino acids of significant biosynthetic interest ............................................................. 57 2.2.2: Specific aims ....................................................................... 62 2.3: Results…… ...................................................................................... 63 2.2.1: Molecular basis for the biosynthesis of the cyclomarins ...... 63 2.2.2: 2-Amino-3,5-dimethyl-4-hexenoic acid (ADH) biosynthesis 69 2.2.3: Reverse N-prenylation of the tryptophan residue ................ 73 2.2.4: Oxidative tailoring ................................................................ 76 2.4: Discussion….. ................................................................................ …79 2.5: Experimental section ......................................................................... 83 2.5.1: General experimental procedures ....................................... 83 2.5.2: Bacterial strains, plasmids, culture conditions, and DNA manipulations ....................................................................... 84 2.5.3: Cultivation of S arenicola and isolation of 1, 8, and 9 .......... 85 2.5.4: Labeling experiments .......................................................... 87 2.5.5: DNA sequence analysis of the cyclomarin gene cluster (cym) ........................................................................ 87 vi 2.5.6: Conjugation protocol for S. arenicola .................................. 88 2.5.7: Inactivation of cymD, cymQ, cymS, cymV and cym W ........ 88 2.5.8: Analysis of the S. arenicola cym mutants ............................ 89 2.6: Appendix…… .................................................................................... 90 2.6.1: Phylogenetic analysis of the cytochrome P450s CymO, CymS, CymV to predict their role in cyclomarin and cyclomarazine biosynthesis ................................................. 90 2.7: References ..................................................................................... 120 2.8: Acknowledgements ......................................................................... 125 Chapter 3: Functional Characterization of the Cyclomarin/Cyclomarazine Prenyltransferase CymD Directs the Biosynthesis of Unnatural Cyclic Peptides .......................................................................................................... 126 3.1: Abstract…… ................................................................................... 127 3.2: Introduction ..................................................................................... 127 3.3: Results…….. ................................................................................... 132 3.4: Discussion……. .............................................................................. 137 3.5: Experimental section….. ................................................................. 138 3.5.1: General experimental procedures…. ................................ 138 3.5.2: Bacterial strains and culture conditions ............................. 138 3.5.3: CymD purification .............................................................. 139 3.5.4: CymD prenyltransferase assay ......................................... 140 3.5.5: Purification of N-(1,1-dimethyl-1-allyl)-tryptophan from S. arenicola CNS-205 ............................................................ 141 3.5.6: Chemical complementation of the cymD- mutant and the production of novel cyclomarin and cyclomarazine analogues via mutasynthesis ............................................. 142 3.6 Appendix ..................................................................................................... 144 3.7: References ................................................................................................ 160 3.8: Acknowledgment ....................................................................................... 162 Chapter 4: Efforts towards Functional Characterization and Biosynthetic Manipulation of the 2-Amino-3,5-dimethyl-4-hexenoic Acid Residue of Cyclomarin .......................................................................... 163
Load more

Recommended publications
  • Review of Oxepine-Pyrimidinone-Ketopiperazine Type Nonribosomal Peptides
    H OH metabolites OH Review Review of Oxepine-Pyrimidinone-Ketopiperazine Type Nonribosomal Peptides Yaojie Guo , Jens C. Frisvad and Thomas O. Larsen * Department of Biotechnology and Biomedicine, Technical University of Denmark, Søltofts Plads, Building 221, DK-2800 Kgs. Lyngby, Denmark; [email protected] (Y.G.); [email protected] (J.C.F.) * Correspondence: [email protected]; Tel.: +45-4525-2632 Received: 12 May 2020; Accepted: 8 June 2020; Published: 15 June 2020 Abstract: Recently, a rare class of nonribosomal peptides (NRPs) bearing a unique Oxepine-Pyrimidinone-Ketopiperazine (OPK) scaffold has been exclusively isolated from fungal sources. Based on the number of rings and conjugation systems on the backbone, it can be further categorized into three types A, B, and C. These compounds have been applied to various bioassays, and some have exhibited promising bioactivities like antifungal activity against phytopathogenic fungi and transcriptional activation on liver X receptor α. This review summarizes all the research related to natural OPK NRPs, including their biological sources, chemical structures, bioassays, as well as proposed biosynthetic mechanisms from 1988 to March 2020. The taxonomy of the fungal sources and chirality-related issues of these products are also discussed. Keywords: oxepine; nonribosomal peptides; bioactivity; biosynthesis; fungi; Aspergillus 1. Introduction Nonribosomal peptides (NRPs), mostly found in bacteria and fungi, are a class of peptidyl secondary metabolites biosynthesized by large modularly organized multienzyme complexes named nonribosomal peptide synthetases (NRPSs) [1]. These products are amongst the most structurally diverse secondary metabolites in nature; they exhibit a broad range of activities, which have been exploited in treatments such as the immunosuppressant cyclosporine A and the antibiotic daptomycin [2,3].
    [Show full text]
  • Amide Bond Formation in Nonribosomal Peptide Synthesis
    Amide Bond Formation in Nonribosomal Peptide Synthesis: The Formylation and Condensation Domains Dissertation zur Erlangung des Doktorgrades der Naturwissenschaften (Dr. rer. nat.) dem Fachbereich Chemie der Philipps-Universität Marburg vorgelegt von Georg Schönafinger aus München Marburg/Lahn 2007 Vom Fachbereich Chemie der Philipps-Universität Marburg als Dissertation am _______________ angenommen. Erstgutachter : Prof. Dr. M. A. Marahiel (Philipps-Universität, Marburg) Zweitgutachter : Prof. Dr. L.-O. Essen (Philipps-Universität, Marburg) Tag der Disputation: 20. Dezember 2007 2 The majority of the work presented here has been published: Samel, S.A.†, Schoenafinger, G. †, Knappe, T.A., Marahiel, M.A., Essen, L.O. 2007. Structural and functional insights into a peptide bond-forming bidomain from a nonribosomal peptide synthetase. Structure 15(7): 781-92. † equal contribution Schoenafinger, G., Schracke, N., Linne, U., Marahiel, M.A. 2006. Formylation domain: an essential modifying enzyme for the nonribosomal biosynthesis of linear gramicidin. J Am Chem Soc. 128(23): 7406-7. Schoenafinger, G., Marahiel, M.A. accepted 2007. Nonribosomal Peptides. Wiley Encyclopaedia of Chemical Biology. In press. 3 To Anke 4 Summary Nonribosomal peptides are of outstanding pharmacological interest, since many representatives of this highly diverse class of natural products exhibit therapeutically important activities, such as antibacterial, antitumor and immunosuppressive properties. Understanding their biosynthesis performed by multimodular mega-enzymes, the nonribosomal peptide synthetases (NRPSs), is one of the key determinants in order to be able to reprogram these machineries for the production of novel therapeutics. The central structural motif of all peptides is the peptide (or amide) bond. In this work, two different amide bond forming catalytic entities from NRPSs were studied: The condensation (C) and formylation (F) domains.
    [Show full text]
  • (12) Patent Application Publication (10) Pub. No.: US 2007/0143878 A1 Bhat Et Al
    US 20070143878A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2007/0143878 A1 Bhat et al. (43) Pub. Date: Jun. 21, 2007 (54) NUCLEC ACID MOLECULES AND OTHER of application No. 09/198.779, filed on Nov. 24, 1998, MOLECULES ASSOCATED WITH THE now abandoned. TOCOPHEROL PATHWAY Said application No. 09/233,218 is a continuation-in part of application No. 09/227,586, filed on Jan. 8, (76) Inventors: Barkur G. Bhat, St. Louis, MO (US); 1999, now abandoned. Sekhar S. Boddupalli, Manchester, MO Said application No. 09/233,218 is a continuation-in (US); Ganesh M. Kishore, Creve part of application No. 09/229,413, filed on Jan. 12, Coeur, MO (US); Jingdong Liu, 1999, now abandoned. Ballwin, MO (US); Shaukat H. Rangwala, Ballwin, MO (US); (60) Provisional application No. 60/067,000, filed on Nov. Mylavarapu Venkatramesh, Ballwin, 24, 1997. Provisional application No. 60/066,873, MO (US) filed on Nov. 25, 1997. Provisional application No. 60/069.472, filed on Dec. 9, 1997. Provisional appli Correspondence Address: cation No. 60/074,201, filed on Feb. 10, 1998. Pro ARNOLD & PORTER, LLP visional application No. 60/074.282, filed on Feb. 10, 555 TWELFTH STREET, N.W. 1998. Provisional application No. 60/074,280, filed ATTN IP DOCKETING on Feb. 10, 1998. Provisional application No. 60/074, WASHINGTON, DC 20004 (US) 281, filed on Feb. 10, 1998. Provisional application No. 60/074,566, filed on Feb. 12, 1998. Provisional (21) Appl. No.: 11/329,160 application No. 60/074,567, filed on Feb. 12, 1998.
    [Show full text]
  • Protective Effects of Berberis Crataegina DC. (Ranunculales: Berberidaceae) Extract on Bleomycin-Induced Toxicity in Fruit Flies (Diptera: Drosophilidae)
    Revista de la Sociedad Entomológica Argentina ISSN: 0373-5680 ISSN: 1851-7471 [email protected] Sociedad Entomológica Argentina Argentina Protective effects of Berberis crataegina DC. (Ranunculales: Berberidaceae) extract on Bleomycin-induced toxicity in fruit flies (Diptera: Drosophilidae) ATICI, Tuğba; ALTUN ÇOLAK, Deniz; ERSÖZ, Çağla Protective effects of Berberis crataegina DC. (Ranunculales: Berberidaceae) extract on Bleomycin-induced toxicity in fruit flies (Diptera: Drosophilidae) Revista de la Sociedad Entomológica Argentina, vol. 77, no. 4, 2018 Sociedad Entomológica Argentina, Argentina Available in: https://www.redalyc.org/articulo.oa?id=322055706003 PDF generated from XML JATS4R by Redalyc Project academic non-profit, developed under the open access initiative Artículos Protective effects of Berberis crataegina DC. (Ranunculales: Berberidaceae) extract on Bleomycin-induced toxicity in fruit flies (Diptera: Drosophilidae) Efectos protectores de extracto de Berberis crataegina DC. (Ranunculales: Berberidaceae) frente a toxicidad inducida con Bleomicina en moscas de la fruta (Diptera: Drosophilidae) Tuğba ATICI Institute of Natural and Applied Science, Erzincan Binali Yıldırım University, Turquía Deniz ALTUN ÇOLAK [email protected] Faculty of Art and Science, Erzincan Binali Yıldırım University, Turquía Çağla ERSÖZ Revista de la Sociedad Entomológica Institute of Natural and Applied Science, Erzincan Binali Yıldırım Argentina, vol. 77, no. 4, 2018 University, Turquía Sociedad Entomológica Argentina, Argentina Received: 02 October 2018 Accepted: 06 December 2018 Published: 27 December 2018 Abstract: Antineoplastic agents due to their cytotoxic effects increase the amount of free radical in cells causing cell death. e aims of this study were to assess the possible Redalyc: https://www.redalyc.org/ toxic effects of Bleomycin (BLM) on the number of offspring and survival rate of fruit articulo.oa?id=322055706003 flies as well as the protective role of Berberis crataegina DC.
    [Show full text]
  • Colonization of Lettuce Rhizosphere and Roots by Tagged Streptomyces
    PhD School of Chemistry, Biochemistry and Ecology of Pesticide (CBEA) DOCTORAL DISSERTATION Tagging biocontrol Streptomyces to study lettuce colonization Xiaoyulong Chen To be presented with the permission of the examining board, 28th cycle of the CBEA PhD school, for public examination at Aula Magna C03 (Room C03), Faculty of Agricultural and food sciences, University of Milan, on 17.12.2015. Cover figure: Colonization of lettuce root by Streptomyces cyaneus ZEA17I, examined by confocal laser scanning microscopy. Supervisor: Professor Paolo Cortesi Department of Food, Environmental, and Nutritional Sciences University of Milan, Italy Co-supervisor: Professor Marco Saracchi Department of Food, Environmental, and Nutritional Sciences University of Milan, Italy Coordinator: Professor Daniele Daffonchio Department of Food, Environmental, and Nutritional Sciences University of Milan, Italy Examination board (ordered by first letter of the family name): 1. Professor Paolo Bertolini Department of Agricultural Sciences University of Bologna, Italy 2. Dr. Jonas Bengtsson Department of Zoology Stockholm University, Sweden 3. Professor Daniele Daffonchio Department of Food, Environmental, and Nutritional Sciences University of Milan, Italy 4. Professor George Tsiamis Dep. of Environmental and Nutritional Resources Management University of Patras, Greece 5. Dr. Laura Polito Institute of Molecular Science and Technology, CNR, Italy Contents List of original publications......................................................................................................
    [Show full text]
  • Construction of Hybrid Peptide Synthetases for the Production of A-L-Aspartyl-L-Phenylalanine, a Precursor for the High-Intensity Sweetener Aspartame
    Eur. J. Biochem. 270, 4555–4563 (2003) Ó FEBS 2003 doi:10.1046/j.1432-1033.2003.03858.x Construction of hybrid peptide synthetases for the production of a-L-aspartyl-L-phenylalanine, a precursor for the high-intensity sweetener aspartame Thomas Duerfahrt1, Sascha Doekel1,*, Theo Sonke2, Peter J. L. M. Quaedflieg2 and Mohamed A. Marahiel1 1Philipps-Universita¨t Marburg, Fachbereich Chemie/Biochemie, Marburg, Germany; 2DSM Research, Life Sciences – Advanced Synthesis, Catalysis and Development, Geleen, the Netherlands Microorganisms produce a large number of pharmaco- Product release was ensured by a C-terminally fused logically and biotechnologically important peptides by thioesterase domains and quantified by HPLC/MS ana- using nonribosomal peptide synthetases (NRPSs). Due to lysis. Significant differences of enzyme activity caused by their modular arrangement and their domain organization the fusion strategies were observed. Two forms of the NRPSs are particularly suitable for engineering recom- Asp-Phe dipeptide were detected, the expected a-Asp-Phe binant proteins for the production of novel peptides with and the by-product b-Asp-Phe. Dependent on the turn- interesting properties. In order to compare different over rates ranging from 0.01–0.7 min)1, the amount of strategies of domain assembling and module fusions we a-Asp-Phe was between 75 and 100% of overall product, focused on the selective construction of a set of peptide indicating a direct correlation between the turnover synthetases that catalyze the formation of the dipeptide numbers and the ratios of a-Asp-Phe to b-Asp-Phe. a-L-aspartyl-L-phenylalanine (Asp-Phe), the precursor of Taken together these results provide useful guidelines for the high-intensity sweetener a-L-aspartyl-L-phenylalanine the rational construction of hybrid peptide synthetases.
    [Show full text]
  • 102 4. Biosynthesis of Natural Products Derived from Shikimic Acid
    102 4. Biosynthesis of Natural Products Derived from Shikimic Acid 4.1. Phenyl-Propanoid Natural Products (C6-C3) The biosynthesis of the aromatic amino acids occurs through the shikimic acid pathway, which is found in plants and microorganisms (but not in animals). We (humans) require these amino acids in our diet, since we are unable to produce them. For this reason, molecules that can inhibit enzymes on the shikimate pathway are potentially useful as antibiotics or herbicides, since they should not be toxic for humans. COO COO NH R = H Phenylalanine 3 R = OH Tyrosine R NH3 N Tryptophan H The aromatic amino acids also serve as starting materials for the biosynthesis of many interesting natural products. Here we will focus on the so-called phenyl-propanoide (C6-C3) natural products, e.g.: OH OH OH HO O HO OH HO O Chalcone OH O a Flavone OH O OH O a Flavonone OH OH Ar RO O O O HO O O OH O OR OH Anthocyanine OH O a Flavonol Podophyllotoxin MeO OMe OMe OH COOH Cinnamyl alcohol HO O O Cinnamic acid OH (Zimtsäure) Umbellierfone OH a Coumarin) MeO OH O COOH HO Polymerization OH Wood OH HO OH O OH MeO OMe Shikimic acid O HO 4.2. Shikimic acid biosynthesis The shikimic acid pathway starts in carbohydrate metabolism. Given the great social and industrial significance of this pathway, the enzymes have been intensively investigated. Here we will focus on the mechanisms of action of several key enzymes in the pathway. The following Scheme shows the pathway to shikimic acid: 103 COO- COO- Phosphoenolpyruvate HO COO- 2- O O3P-O 2- O3P-O DHQ-Synthase
    [Show full text]
  • 8.2 Shikimic Acid Pathway
    CHAPTER 8 © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FORAromatic SALE OR DISTRIBUTION and NOT FOR SALE OR DISTRIBUTION Phenolic Compounds © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION CHAPTER OUTLINE Overview Synthesis and Properties of Polyketides 8.1 8.5 Synthesis of Chalcones © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC 8.2 Shikimic Acid Pathway Synthesis of Flavanones and Derivatives NOT FOR SALE ORPhenylalanine DISTRIBUTION and Tyrosine Synthesis NOT FOR SALESynthesis OR DISTRIBUTION and Properties of Flavones Tryptophan Synthesis Synthesis and Properties of Anthocyanidins Synthesis and Properties of Isofl avonoids Phenylpropanoid Pathway 8.3 Examples of Other Plant Polyketide Synthases Synthesis of Trans-Cinnamic Acid Synthesis and Activity of Coumarins Lignin Synthesis Polymerization© Jonesof Monolignols & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC Genetic EngineeringNOT FOR of Lignin SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION Natural Products Derived from the 8.4 Phenylpropanoid Pathway Natural Products from Monolignols © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION © Jones & Bartlett Learning, LLC © Jones & Bartlett Learning, LLC NOT FOR SALE OR DISTRIBUTION NOT FOR SALE OR DISTRIBUTION 119 © Jones & Bartlett Learning, LLC.
    [Show full text]
  • Nonribosomal Peptides Synthetases and Their Applications in Industry Mario Alberto Martínez‑Núñez1,2* and Víctor Eric López Y López3
    Martínez‑Núñez and López Sustain Chem Process (2016) 4:13 DOI 10.1186/s40508-016-0057-6 REVIEW Open Access Nonribosomal peptides synthetases and their applications in industry Mario Alberto Martínez‑Núñez1,2* and Víctor Eric López y López3 Abstract Nonribosomal peptides are products that fall into the class of secondary metabolites with a diverse properties as toxins, siderophores, pigments, or antibiotics, among others. Unlike other proteins, its biosynthesis is independent of ribosomal machinery. Nonribosomal peptides are synthesized on large nonribosomal peptide synthetase (NRPS) enzyme complexes. NRPSs are defined as multimodular enzymes, consisting of repeated modules. The NRPS enzymes are at operons and their regulation can be positive or negative at transcriptional or post-translational level. The pres‑ ence of NRPS enzymes has been reported in the three domains of life, being prevalent in bacteria. Nonribosomal pep‑ tides are use in human medicine, crop protection, or environment restoration; and their use as commercial products has been approved by the U. S Food and Drug Administration (FDA) and the U. S. Environmental Protection Agency (EPA). The key features of nonribosomal peptides and NRPS enzymes, and some of their applications in industry are summarized. Keywords: Nonribosomal peptides, Nonribosomal peptides synthetases, Environmental restoration, Human healt Background half of the NRPS enzymes found in a genome-mining Nonribosomal peptides is a diverse family of natural study of 2699 genomes by Wang et al. [1] are nonmodular products fall into the class of secondary metabolites with NRPS enzymes. Nonmodular NRPS enzymes are found a diverse properties as toxins, siderophores, pigments, in siderophore biosynthetic pathways like EntE and antibiotics, cytostatics, immunosuppressants or antican- VibH in enterobactin, and VibE in vibriobactin [5] or as cer agents [1, 2]; and have a particularity: their synthesis a stand-alone peptidyl carrier protein such as BlmI from is independent of ribosomal machinery.
    [Show full text]
  • Enhancement of Bleomycin Production in Streptomyces Verticillus Through Global Metabolic Regulation of N-Acetylglucosamine and A
    Chen et al. Microb Cell Fact (2020) 19:32 https://doi.org/10.1186/s12934-020-01301-8 Microbial Cell Factories RESEARCH Open Access Enhancement of bleomycin production in Streptomyces verticillus through global metabolic regulation of N-acetylglucosamine and assisted metabolic profling analysis Hong Chen1,2†, Jiaqi Cui1,2†, Pan Wang1,2, Xin Wang1,2 and Jianping Wen1,2* Abstract Background: Bleomycin is a broad-spectrum glycopeptide antitumor antibiotic produced by Streptomyces verticil- lus. Clinically, the mixture of bleomycin A2 and bleomycin B2 is widely used in combination with other drugs for the treatment of various cancers. As a secondary metabolite, the biosynthesis of bleomycin is precisely controlled by the complex extra-/intracellular regulation mechanisms, it is imperative to investigate the global metabolic and regula- tory system involved in bleomycin biosynthesis for increasing bleomycin production. Results: N-acetylglucosamine (GlcNAc), the vital signaling molecule controlling the onset of development and antibiotic synthesis in Streptomyces, was found to increase the yields of bleomycins signifcantly in chemically defned medium. To mine the gene information relevant to GlcNAc metabolism, the DNA sequences of dasR-dasA-dasBCD- nagB and nagKA in S. verticillus were determined by chromosome walking. From the results of Real time fuorescence quantitative PCR (RT-qPCR) and electrophoretic mobility shift assays (EMSAs), the repression of the expression of nagB and nagKA by the global regulator DasR was released under induction with GlcNAc. The relief of blmT expres- sion repression by BlmR was the main reason for increased bleomycin production. DasR, however, could not directly afect the expression of the pathway-specifc repressor BlmR in the bleomycins gene cluster.
    [Show full text]
  • Bactrev00065-0077.Pdf
    BACnEIUOLOGICAL REVIEWS, Dec. 1968, p. 465-492 Vol. 32, No. 4, Pt. 2 Copyright © 1968 American Society for Microbiology Printed in U.S.A. Pathways of Biosynthesis of Aromatic Amino Acids and Vitamins and Their Control in Microorganisms FRANK GIBSON AND JAMES PITTARD John Curtin School of Medical Research, Australian National University, Canberra, Australia, and School of Microbiology, University of Melbourne, Australia INTRODUCTION................................................................ 465 INTERMEDIATES IN AROMATIC BIOsYNTHESIS ...................................... 466 Common Pathway ........................................................... 466 Tryptophan Pathway ........................................................ 468 Pathways to Phenylalanine and Tyrosine ........................................ 469 Pathway to 4-Aminobenzoic Acid.............................................. 469 Intermediates in Ubiquinone Biosynthesis ....................................... 470 Intermediates in Vitamin K Biosynthesis ........................................ 471 Pathways Involving 2,3-Dihydroxybenzoate ..................................... 472 Other Phenolic Growth Factors ............................................... 473 ISOENZYMES AND PROTEIN AGGREGATES CONCERNED IN AROMATIC BiosYNTHESIS ........ 474 Common Pathway ........................................................... 474 Tryptophan Pathway ......................................................... 474 Phenylalanine and Tyrosine Pathways .........................................
    [Show full text]
  • Exploring the Potential of Antibiotic Production from Rare Actinobacteria by Whole-Genome Sequencing and Guided MS/MS Analysis
    fmicb-11-01540 July 27, 2020 Time: 14:51 # 1 ORIGINAL RESEARCH published: 15 July 2020 doi: 10.3389/fmicb.2020.01540 Exploring the Potential of Antibiotic Production From Rare Actinobacteria by Whole-Genome Sequencing and Guided MS/MS Analysis Dini Hu1,2, Chenghang Sun3, Tao Jin4, Guangyi Fan4, Kai Meng Mok2, Kai Li1* and Simon Ming-Yuen Lee5* 1 School of Ecology and Nature Conservation, Beijing Forestry University, Beijing, China, 2 Department of Civil and Environmental Engineering, Faculty of Science and Technology, University of Macau, Macau, China, 3 Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China, 4 Beijing Genomics Institute, Shenzhen, China, 5 State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macau, China Actinobacteria are well recognized for their production of structurally diverse bioactive Edited by: secondary metabolites, but the rare actinobacterial genera have been underexploited Sukhwan Yoon, for such potential. To search for new sources of active compounds, an experiment Korea Advanced Institute of Science combining genomic analysis and tandem mass spectrometry (MS/MS) screening and Technology, South Korea was designed to isolate and characterize actinobacterial strains from a mangrove Reviewed by: Hui Li, environment in Macau. Fourteen actinobacterial strains were isolated from the collected Jinan University, China samples. Partial 16S sequences indicated that they were from six genera, including Baogang Zhang, China University of Geosciences, Brevibacterium, Curtobacterium, Kineococcus, Micromonospora, Mycobacterium, and China Streptomyces. The isolate sp.01 showing 99.28% sequence similarity with a reference *Correspondence: rare actinobacterial species Micromonospora aurantiaca ATCC 27029T was selected for Kai Li whole genome sequencing.
    [Show full text]