DOCSLIB.ORG

Chapter 4 Engineering of Streptomyces Lividans For

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

Engineering of Streptomyces albus J1074 and Streptomyces lividans TK24 for Natural Products Production Dissertation zur Erlangung des Grades des Doktors der Naturwissenschaften der Naturwissenschaftlich-Technischen Fakultät der Universität des Saarlandes Von Yousra Ahmed Saarbrücken 2019 2 Tag des Kolloquiums: 09. August 2019 Dekan: Prof. Dr. rer. nat. Guido Kickelbick Berichterstatter: Prof. Dr. Andriy Luzhetskyy Prof. Dr. Rolf Müller Vorsitz: Prof. Dr. Volkhard Helms Akad. Mitarbeiter: Dr. Stefan Boettcher 3 Diese Arbeit entstand unter der Anleitung von Prof. Dr. Andriy Luzhetskyy in der Fachrichtung Pharmazeutische Biotechnologie der Naturwissenschaftlich-Technischen Fakultät der Universität des Saarlandes von Mai 2013 bis August 2018. 4 Acknowledgement Acknowledgments Firstly, I would like to express my deep sense of gratitude and respectful regards to my supervisor Prof. Dr. Andriy Luzhetskyy for giving me the opportunity to work in his group on these exciting projects. I highly appreciate his invaluable guidance and continuous support throughout my PhD studies. I am also thankful to him for reading the manuscript of this thesis and for his recommendations to improve it. I would like to thank Prof. Dr. Rolf Müller for his insightful comments and suggestions during the committee meetings and for being the second reviewer of this thesis. I am forever grateful to Dr. Yuriy Rebets, my lab supervisor, for his patience, motivation, and for sparing his valuable time whenever I needed. His useful experimental advice, constructive suggestions and scientific inspiration made it possible for me to achieve my goal. I am indebted to him for his help, suggestions and correction of the entire thesis. It has been an absolute pleasure to work with and learn from him. I am also very grateful to all my colleagues in AMEG group for their wonderful collaboration. They supported me impressively and were always willing to help me. Thanks a lot for the nice time we have spent together outside of work. I will cherish these memories forever. I would like to express my deepest gratitude to my parents, Samaa and MhD. Saleh, my sister Souhaila and my brothers: Ossama, Ahmed, Mouwafak and Abderrahman, who always believe in me and encourage me to follow my dreams. Words fail to express my appreciation and respect to them for their moral and emotional support in all aspects of my life. I would like to thank my nieces and nephews that made me laugh from my heart in the hardest moments. To Mouwafak and Abderrahman I am so grateful to have you here beside me and for helping in whatever way you could during all challenging times. Thanks a lot for the very delicious meals and for making sure that I did not have to cook when I was very busy. I am also obliged to my other family members and friends who have supported me along the way. I sincerely express my gratitude from my heart to my beloved husband, Ahmad, for his warm love, continued patience, and endless support during the happy and hard times. He has been a true and great supporter for the last two and half year of my studies, living every single minute of it. I thank God for enlightening my life with his presence. Without him this journey would not have been possible. Yousra Ahmed Saarbrücken, April 01, 2019 5 Publications and Conference Contributions Publications Amin, R., Franz-Wachtel, M., Tiffert, Y., Heberer, M., Meky, M., Ahmed, Y., Matthews, A., Krysenko, S., Jakobi, M., Hinder, M., et al. (2016) Post- translational Serine/Threonine Phosphorylation and Lysine Acetylation: A Novel Regulatory Aspect of the Global Nitrogen Response Regulator GlnR in S. coelicolor M145. Front. Mol. Biosci., 3, 38 Ahmed, Y., Rebets, Y., Tokovenko, B., Brötz, E., Luzhetskyy, A. 2017. Identification of butenolide regulatory system controlling secondary metabolism in Streptomyces albus J1074. Sci Rep 7:9784. Busche, T., Tsolis, K.C., Koepff, J., Rebets Y., Ruckert, C., Hamed, M.B., Bleidt, A., Wiechert, W., Lopatniuk, M., Ahmed, Y. et al. Multi-Omics and Targeted Approaches to Determine the Role of Cellular Proteases in Streptomyces Protein Secretion. Front Microbiol 2018, 9:1174. Rebets, Y., Tsolis, K.C., Guðmundsdóttir, E.E., Koepff, J., Wawiernia, B., Busche, T., Bleidt, A., Horbal, L., Myronovskyi, M., Ahmed, Y., Wiechert, W., Rückert, C., Hamed, M.B., Bilyk, B., Anné, J., Friðjónsson, Ó., Kalinowski, J., Oldiges, M., Economou, A., Luzhetskyy, A. Characterization of Sigma Factor Genes in Streptomyces lividans TK24 Using a Genomic Library-Based Approach for Multiple Gene Deletions. Frontiers in Microbiology 9, doi:10.3389/fmicb.2018.03033 (2018). Ahmed, Y., Rebets, Y., Rodríguez Estévez, M., Luzhetskyy, A. Engineering of Streptomyces lividans for heterologous expression of secondary metabolites gene clusters (manuscript in preparation) Conference Contributions Amin, R., Tiffert, Y., Heberer, M., Ahmed, Y., Krysenko, S., Jakobi, M., Hinder, M., et al. Post-translational Serine/Threonine Phosphorylation and Lysine Acetylation: A Novel Regulatory Aspect of the Global Nitrogen Response Regulator GlnR in S. coelicolor M145, the Annual Meeting of the part "Association for General and Applied Microbiology“ Tuebingen, Germany (VAAM). (Poster) Ahmed, Y., Rebets, Y., Brötz, E. and Luzhetskyy, A. new strategy in awaking silent secondary metabolites biosynthesis gene clusters in Streptomyces albus J1074, VAAM workshop, Freiburg, Germany 2014. (Poster) Ahmed, Y., Rebets, Y., Brötz, E. and Luzhetskyy, A. Regulatory elements silencing the secondary metabolism in Streptomyces albus J1074. 5th HIPS Symposium on Pharmaceutical Sciences Devoted to Infection Research, Saarbruecken, 2015. (Poster) 6 Abstract Abstract Actinobacteria have remarkable chemical potential that is not explored due to low level of corresponding genes expression. In order to uncover this reservoir of natural products we developed a reporter-guided screening strategy combined with transposon mutagenesis. It was used to activate silent polycyclic tetramate macrolactam biosynthesis gene cluster in Streptomyces albus J1074. As result, the mutant with awaken secondary metabolism was obtained. Analysis of this strain led to identification of new regulatory system consisting of transcriptional regulator XNR_3174 and bacterial hormone-like compound butenolide. XNR_3174 and butenolide biosynthesis genes orthologues are present in the genomes of different Streptomyces. The identified regulatory system comprises a new condition-depended cascade controlling secondary metabolism in Actinobacteria. We also developed new host strains for heterologous production of natural products by deleting 11 endogenous secondary metabolite gene clusters from chromosome of S. lividans and introducing up to 2 sites for integration of foreign DNA. When expressing three heterologous gene clusters the generated hosts have shown better performance than the parental strain. S. lividans TK24 was also improved as a host for heterologous protein production by deleting a set of proteases encoding genes. The developed strains represent a step forward to a better panel of organisms for bioprospecting and genome mining of novel natural products. 7 Zusammenfassung Zusammenfassung Actinobakterien besitzen ungeahntes chemisches Potenzial, das aufgrund niedriger Exprimierung entsprechender Gene nicht erforscht werden kann. Um Dieses aufzudecken, wurde eine Kombination aus reportergeführter Screening-Strategie und Transposon Mutagene entwickelt. Die Verwendung dieser Strategie führte zur Aktivierung des polyzyklischen Tetramat- Makrolaktam Gen-Clusters in Streptomyces albus J1074. Der erhaltene Stamm weißt eine aktivierte Produktion von Sekundärmetaboliten auf und Analysen führten zur Identifizierung eines Regulationssystems, bestehend aus dem transkriptionellen Regulator XNR_3174 und dem hormonähnlichen Naturstoff Butenolid. Orthologe von XNR_3174 und der Butenolid Gene findet man in Genomen verschiedener Streptomyceten. Das identifizierte System umfasst die neue „condition-dependent“-Kaskade, die den Sekundärstoffwechsel in Actinobakterien steuert. Zusätzlich entwickelten wir, durch das Entfernen von 11 endogenen Gen-Clustern aus dem Genom von S. lividans und dem Einfügen von zwei DNA Integrationsstellen, neue Wirtsstämme für die heterologe Produktion von Naturstoffen. Bei der heterologen Exprimierung von drei Gen- Clustern zeigten die optimierten Wirte bessere Resultate als der ursprüngliche Stamm. Durch das Entfernen der Proteasegene wurde S. lividans TK24 als Wirt für die Proteinproduktion verbessert. Die entwickelten Stämme vereinfachen die Bioprospektion und die Entdeckung neuer Naturstoffe unter Verwendung des „Genome-Mining“ Ansatzes. 8 Table of Contents Table of Contents Acknowledgment 5 Publications and Conference Contributions 6 Abstract 7 Zusammenfassung 8 Table of Contents 9 1. Introduction 11 1.1 Importance of Natural Products in Medicines Development 11 1.2 Natural Products of Actinobacteria 14 1.3 Activation of Silent Biosynthetic Gene Clusters 15 1.3.1 OSMAC: One Strain Many Compounds 16 1.3.2 Manipulation of Transcriptional and Translational Machineries 18 1.3.3 Manipulation of Global and Pathway-Specific Regulators 19 a) Manipulation of global Regulators 20 b) Manipulation of Pathway-Specific Regulators 21 1.3.4 Gene Cluster Refactoring 23 1.3.5 Reporter-Guided Mutant Selection (RGMS) 25 1.4 Heterologous Expression of Secondary Metabolite Gene Clusters 27 1.4.1 Streptomyces coelicolor 28 1.4.2 Streptomyces avermitilis 29 1.4.3 Streptomyces albus 29 1.4.4 Streptomyces lividans 30 1.5
Recommended publications
  • Successful Drug Discovery Informed by Actinobacterial Systematics

    Successful Drug Discovery Informed by Actinobacterial Systematics

    Successful Drug Discovery Informed by Actinobacterial Systematics Verrucosispora HPLC-DAD analysis of culture filtrate Structures of Abyssomicins Biological activity T DAD1, 7.382 (196 mAU,Up2) of 002-0101.D V. maris AB-18-032 mAU CH3 CH3 T extract H3C H3C Antibacterial activity (MIC): S. leeuwenhoekii C34 maris AB-18-032 175 mAU DAD1 A, Sig=210,10 150 C DAD1 B, Sig=230,10 O O DAD1 C, Sig=260,20 125 7 7 500 Rt 7.4 min DAD1 D, Sig=280,20 O O O O Growth inhibition of Gram-positive bacteria DAD1 , Sig=310,20 100 Abyssomicins DAD1 F, Sig=360,40 C 75 DAD1 G, Sig=435,40 Staphylococcus aureus (MRSA) 4 µg/ml DAD1 H, Sig=500,40 50 400 O O 25 O O Staphylococcus aureus (iVRSA) 13 µg/ml 0 CH CH3 300 400 500 nm 3 DAD1, 7.446 (300 mAU,Dn1) of 002-0101.D 300 mAU Mode of action: C HO atrop-C HO 250 atrop-C CH3 CH3 CH3 CH3 200 H C H C H C inhibitior of pABA biosynthesis 200 Rt 7.5 min H3C 3 3 3 Proximicin A Proximicin 150 HO O HO O O O O O O O O O A 100 O covalent binding to Cys263 of PabB 100 N 50 O O HO O O Sea of Japan B O O N O O (4-amino-4-deoxychorismate synthase) by 0 CH CH3 CH3 CH3 3 300 400 500 nm HO HO HO HO Michael addition -289 m 0 B D G H 2 4 6 8 10 12 14 16 min Newcastle Michael Goodfellow, School of Biology, University Newcastle University, Newcastle upon Tyne Atacama Desert In This Talk I will Consider: • Actinobacteria as a key group in the search for new therapeutic drugs.
  • Genome-Based Classification of The

    Genome-Based Classification of The

    Antonie van Leeuwenhoek https://doi.org/10.1007/s10482-021-01564-0 (0123456789().,-volV)( 0123456789().,-volV) ORIGINAL PAPER Genome-based classification of the Streptomyces violaceusniger clade and description of Streptomyces sabulosicollis sp. nov. from an Indonesian sand dune Ali B. Kusuma . Imen Nouioui . Michael Goodfellow Received: 26 January 2021 / Accepted: 18 March 2021 Ó The Author(s) 2021 Abstract A polyphasic study was designed to fatty acids. Whole-genome sequences generated for determine the taxonomic provenance of a strain, the isolate and Streptomyces albiflaviniger DSM isolate PRKS01-29T, recovered from an Indonesian 41598T and Streptomyces javensis DSM 41764T were sand dune and provisionally assigned to the Strepto- compared with phylogenetically closely related myces violaceusniger clade. Genomic, genotypic and strains, the isolate formed a branch within the S. phenotypic data confirmed this classification. The violaceusniger clade in the resultant phylogenomic isolate formed an extensively branched substrate tree. Whole-genome sequences data showed that mycelium which carried aerial hyphae that differen- isolate PRKS01-29T was most closely related to the tiated into spiral chains of rugose ornamented spores, S. albiflaviniger strain but was distinguished from the contained LL-as the wall diaminopimelic acid, MK-9 latter and from other members of the clade using (H6,H8) as predominant isoprenologues, phos- combinations of phenotypic properties and average phatidylethanolamine as the diagnostic phospholipid nucleotide identity and digital DNA:DNA hybridiza- and major proportions of saturated, iso- and anteiso- tion scores. Consequently, it is proposed that isolate PRKS01-29T (= CCMM B1303T = ICEBB-02T- = NCIMB 15210T) should be classified in the genus Supplementary Information The online version contains Streptomyces as Streptomyces sabulosicollis sp.
  • Alpine Soil Bacterial Community and Environmental Filters Bahar Shahnavaz

    Alpine Soil Bacterial Community and Environmental Filters Bahar Shahnavaz

    Alpine soil bacterial community and environmental filters Bahar Shahnavaz To cite this version: Bahar Shahnavaz. Alpine soil bacterial community and environmental filters. Other [q-bio.OT]. Université Joseph-Fourier - Grenoble I, 2009. English. tel-00515414 HAL Id: tel-00515414 https://tel.archives-ouvertes.fr/tel-00515414 Submitted on 6 Sep 2010 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. THÈSE Pour l’obtention du titre de l'Université Joseph-Fourier - Grenoble 1 École Doctorale : Chimie et Sciences du Vivant Spécialité : Biodiversité, Écologie, Environnement Communautés bactériennes de sols alpins et filtres environnementaux Par Bahar SHAHNAVAZ Soutenue devant jury le 25 Septembre 2009 Composition du jury Dr. Thierry HEULIN Rapporteur Dr. Christian JEANTHON Rapporteur Dr. Sylvie NAZARET Examinateur Dr. Jean MARTIN Examinateur Dr. Yves JOUANNEAU Président du jury Dr. Roberto GEREMIA Directeur de thèse Thèse préparée au sien du Laboratoire d’Ecologie Alpine (LECA, UMR UJF- CNRS 5553) THÈSE Pour l’obtention du titre de Docteur de l’Université de Grenoble École Doctorale : Chimie et Sciences du Vivant Spécialité : Biodiversité, Écologie, Environnement Communautés bactériennes de sols alpins et filtres environnementaux Bahar SHAHNAVAZ Directeur : Roberto GEREMIA Soutenue devant jury le 25 Septembre 2009 Composition du jury Dr.
  • Genomic and Phylogenomic Insights Into the Family Streptomycetaceae Lead

    Genomic and Phylogenomic Insights Into the Family Streptomycetaceae Lead

    1 Supplementary Material 2 Genomic and phylogenomic insights into the family Streptomycetaceae lead 3 to proposal of Charcoactinosporaceae fam. nov. and 8 novel genera with 4 emended descriptions of Streptomyces calvus 5 Munusamy Madhaiyan1, †, *, Venkatakrishnan Sivaraj Saravanan2, †, Wah-Seng See-Too3, † 6 1Temasek Life Sciences Laboratory, 1 Research Link, National University of Singapore, 7 Singapore 117604; 2Department of Microbiology, Indira Gandhi College of Arts and Science, 8 Kathirkamam 605009, Pondicherry, India; 3Division of Genetics and Molecular Biology, 9 Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, 10 Malaysia 1 11 Table S3. List of the core genes in the genome used for phylogenomic analysis. NCBI Protein Accession Gene WP_074993204.1 NUDIX hydrolase WP_070028582.1 YggS family pyridoxal phosphate-dependent enzyme WP_074992763.1 ParB/RepB/Spo0J family partition protein WP_070022023.1 lipoyl(octanoyl) transferase LipB WP_070025151.1 FABP family protein WP_070027039.1 heat-inducible transcriptional repressor HrcA WP_074992865.1 folate-binding protein YgfZ WP_074992658.1 recombination protein RecR WP_074991826.1 HIT domain-containing protein WP_070024163.1 adenylosuccinate synthase WP_009190566.1 anti-sigma regulatory factor WP_071828679.1 preprotein translocase subunit SecG WP_070026304.1 50S ribosomal protein L13 WP_009190144.1 30S ribosomal protein S5 WP_014674378.1 30S ribosomal protein S8 WP_070026314.1 50S ribosomal protein L5 WP_009300593.1 30S ribosomal protein S13 WP_003998809.1
  • Streptomyces Cytochrome P450 Enzymes and Their Roles in the Biosynthesis of Macrolide Therapeutic Agents

    Streptomyces Cytochrome P450 Enzymes and Their Roles in the Biosynthesis of Macrolide Therapeutic Agents

    Review Biomol Ther 27(2), 127-133 (2019) Streptomyces Cytochrome P450 Enzymes and Their Roles in the Biosynthesis of Macrolide Therapeutic Agents Myung-A Cho, Songhee Han, Young-Ran Lim, Vitchan Kim, Harim Kim and Donghak Kim,* Department of Biological Sciences, Konkuk University, Seoul 05025, Republic of Korea Abstract The study of the genus Streptomyces is of particular interest because it produces a wide array of clinically important bioactive molecules. The genomic sequencing of many Streptomyces species has revealed unusually large numbers of cytochrome P450 genes, which are involved in the biosynthesis of secondary metabolites. Many macrolide biosynthetic pathways are catalyzed by a series of enzymes in gene clusters including polyketide and non-ribosomal peptide synthesis. In general, Streptomyces P450 enzymes accelerate the final, post-polyketide synthesis steps to enhance the structural architecture of macrolide chemistry. In this review, we discuss the major Streptomyces P450 enzymes research focused on the biosynthetic processing of macrolide therapeutic agents, with an emphasis on their biochemical mechanisms and structural insights. Key Words: Streptomyces, P450, CYP, Biosynthesis, Macrolide, Secondary metabolite INTRODUCTION isms became important to human health with the discovery of penicillin in 1928 by Fleming, and the discovery of the anti- The phylum actinobacteria is one of the major lineages cur- tuberculosis agent streptomycin from Streptomyces griseus rently recognized within bacteria (Ventura et al., 2007). Acti- in 1944 by Waksman (Ikeda, 2017). More recently, the 2015 nobacteria are widely distributed in terrestrial, especially soil, Nobel prize in Physiology or Medicine was awarded to Omura and aquatic ecosystems (McCarthy and Williams, 1992; Stach and Campbell for their contributions to the discovery of the and Bull, 2005).
  • Biosynthetic Origin of Anthracimycin: a Tricyclic Macrolide from Streptomyces Sp

    Biosynthetic Origin of Anthracimycin: a Tricyclic Macrolide from Streptomyces Sp

    The Journal of Antibiotics (2016) 69, 403–405 & 2016 Japan Antibiotics Research Association All rights reserved 0021-8820/16 www.nature.com/ja NOTE Biosynthetic origin of anthracimycin: a tricyclic macrolide from Streptomyces sp. Enjuro Harunari1, Hisayuki Komaki2 and Yasuhiro Igarashi1 The Journal of Antibiotics (2016) 69, 403–405; doi:10.1038/ja.2015.118; published online 2 December 2015 Anthracimycin (1) is a decalin-fused tricyclic macrolide isolated from signals arising from the 13C–13C coupling were clearly observed for all terrestrial and marine Streptomyces species (Figure 1).1–3 1 shows the carbons except for three methyl groups (C-23, C-24 and C-25) strong activity against Gram-stain-positive bacteria including Bacillus and C-11 and C-12 (Supplementary Figure S1). The outer split peaks anthracis and methicillin-resistant Staphylococcus aureus (MRSA).3,4 were invisible owing to the roof effect7 caused by the close chemical The most intriguing feature of this molecule is its enantiomerism to shifts between C-11 and C-12 (Δ 0.11 p.p.m.) although these carbons chlorotonil A (2), a secondary metabolite of a myxobacterium are likely derived from a single acetate unit. In the 2D-INADEQUATE 5 1 Sorangium cellulosum (Figure 1). Except for a methyl substitution at spectrum measured with a parameter set optimized for JCC 45 Hz, 13 C-8 in 2, the absolute configurations of the remaining chiral centers at cross peaks derived from the intact C2 acetate units were observed C-2, C-6, C-7, C-12, C-15 and C-16 are opposite between 1 and 2 for C-1/C-2, C-3/C-4, C-5/C-6, C-7/C-8, C-9/C-10, C-13/C-14, C-15/ although the dichrorination at C-4 in 2 is an additional difference C-16, C-17/C-18, C-19/C-20 and C-21/C-22 (Table 1, Supplementary between these compounds.
  • Study of Actinobacteria and Their Secondary Metabolites from Various Habitats in Indonesia and Deep-Sea of the North Atlantic Ocean

    Study of Actinobacteria and Their Secondary Metabolites from Various Habitats in Indonesia and Deep-Sea of the North Atlantic Ocean

    Study of Actinobacteria and their Secondary Metabolites from Various Habitats in Indonesia and Deep-Sea of the North Atlantic Ocean Von der Fakultät für Lebenswissenschaften der Technischen Universität Carolo-Wilhelmina zu Braunschweig zur Erlangung des Grades eines Doktors der Naturwissenschaften (Dr. rer. nat.) genehmigte D i s s e r t a t i o n von Chandra Risdian aus Jakarta / Indonesien 1. Referent: Professor Dr. Michael Steinert 2. Referent: Privatdozent Dr. Joachim M. Wink eingereicht am: 18.12.2019 mündliche Prüfung (Disputation) am: 04.03.2020 Druckjahr 2020 ii Vorveröffentlichungen der Dissertation Teilergebnisse aus dieser Arbeit wurden mit Genehmigung der Fakultät für Lebenswissenschaften, vertreten durch den Mentor der Arbeit, in folgenden Beiträgen vorab veröffentlicht: Publikationen Risdian C, Primahana G, Mozef T, Dewi RT, Ratnakomala S, Lisdiyanti P, and Wink J. Screening of antimicrobial producing Actinobacteria from Enggano Island, Indonesia. AIP Conf Proc 2024(1):020039 (2018). Risdian C, Mozef T, and Wink J. Biosynthesis of polyketides in Streptomyces. Microorganisms 7(5):124 (2019) Posterbeiträge Risdian C, Mozef T, Dewi RT, Primahana G, Lisdiyanti P, Ratnakomala S, Sudarman E, Steinert M, and Wink J. Isolation, characterization, and screening of antibiotic producing Streptomyces spp. collected from soil of Enggano Island, Indonesia. The 7th HIPS Symposium, Saarbrücken, Germany (2017). Risdian C, Ratnakomala S, Lisdiyanti P, Mozef T, and Wink J. Multilocus sequence analysis of Streptomyces sp. SHP 1-2 and related species for phylogenetic and taxonomic studies. The HIPS Symposium, Saarbrücken, Germany (2019). iii Acknowledgements Acknowledgements First and foremost I would like to express my deep gratitude to my mentor PD Dr.
  • Towards the Total Synthesis of Anthracimycin

    Towards the Total Synthesis of Anthracimycin

    Towards the Total Synthesis of Anthracimycin Giacomo Lodovici Doctor of Philosophy University of York Chemistry January 2019 1. Abstract In 2013 W. Fenical et al. reported the isolation of a natural product from a marine microorganism of streptomyces species, which possessed significant activity against Gram-positive pathogens Bacillus anthracis, methicillin-resistant and vancomycin resistant Staphylococcus aureus (MRSA).15 The compound responsible for the antibiotic activity was found to be the 14-membered macrolide named anthracimycin. The research detailed in this thesis describes the efforts towards the total synthesis of this natural product and specifically the formation of the core of anthracimycin in 12-steps. Direct palladium catalysed oxidation formed the enone used as a dienophile in a stereo- and regio-selective Diels‒Alder/epimerisation sequence, which afforded the trans-decalin. A facial and stereoselective Hosomi‒Sakurai 1,4-addition reaction, followed by a selective borylation/dihydroxylation sequence on the exocyclic alkene, allowed the formation of the core of anthracimycin (Scheme 1). Scheme 1. The formation of the decalin core of anthracimcyin. 1 2. Contents 1. Abstract ............................................................................................................................................... 1 2. Contents .............................................................................................................................................. 2 3. List of Figures .....................................................................................................................................
  • Book of Abstracts

    Book of Abstracts

    Book of Abstracts VI International Conference on Environmental, Industrial and Applied Microbiology – BioMicroWorld2015 Barcelona (Spain), 28-30 October 2015 VI International Conference on Environmental, Industrial and Applied Microbiology - BioMicroWorld2015 VI International Conference on Environmental, Industrial and Applied Microbiology - BioMicroWorld2015 Barcelona (Spain), 28-30 October 2015 Barcelona (Spain), 28-30 October 2015 Effect on Metaresistome and metabolic profile (CLPP) soil bacterial communities of different 30 Introduction XXIII agricultural management in Vitis vinifera plots Plenary Lectures XXIX Effects of Inorganic Fertilizers and Organic Manure on Cyanobacteria of Paddy Fields 31 Endophytic bacteria isolated from two varieties of Oryza sativa cultivated in southern Brazil. 32 Session 1: Agriculture, soil, forest microbiology 1 Evaluation of the biological activity of extracts obtained from bacteria associated with nematodes 33 1-Aminocyclopropane-1-carboxylate deaminase producing bacteria promote wheat growth under 2 against Leaf Cutting Ants Atta cephalotes Linnaeus (Hymenoptera Formicidae) water stress Exploration of microorganisms associated with insects, searching for active substances produced 34 454-Pyrosequencing reveals high and differential level of fungal diversity in the Oasis farming 3 from bacteria system in Oman Functional traits of endophytic and rhizosphere fungi and bacteria of Butia archeri Glassman roots 35 A new Cryphonectria hypovirus 1 subtype found in Portugal 4 Genetic tools for site-specific
  • Genomic and Phylogenomic Insights Into the Family Streptomycetaceae Lead to Proposal of Charcoactinosporaceae Fam. Nov. and 8 No

    Genomic and Phylogenomic Insights Into the Family Streptomycetaceae Lead to Proposal of Charcoactinosporaceae Fam. Nov. and 8 No

    bioRxiv preprint doi: https://doi.org/10.1101/2020.07.08.193797; this version posted July 8, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 Genomic and phylogenomic insights into the family Streptomycetaceae 2 lead to proposal of Charcoactinosporaceae fam. nov. and 8 novel genera 3 with emended descriptions of Streptomyces calvus 4 Munusamy Madhaiyan1, †, * Venkatakrishnan Sivaraj Saravanan2, † Wah-Seng See-Too3, † 5 1Temasek Life Sciences Laboratory, 1 Research Link, National University of Singapore, 6 Singapore 117604; 2Department of Microbiology, Indira Gandhi College of Arts and Science, 7 Kathirkamam 605009, Pondicherry, India; 3Division of Genetics and Molecular Biology, 8 Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, 9 Malaysia 10 *Corresponding author: Temasek Life Sciences Laboratory, 1 Research Link, National 11 University of Singapore, Singapore 117604; E-mail: [email protected] 12 †All these authors have contributed equally to this work 13 Abstract 14 Streptomycetaceae is one of the oldest families within phylum Actinobacteria and it is large and 15 diverse in terms of number of described taxa. The members of the family are known for their 16 ability to produce medically important secondary metabolites and antibiotics. In this study, 17 strains showing low 16S rRNA gene similarity (<97.3 %) with other members of 18 Streptomycetaceae were identified and subjected to phylogenomic analysis using 33 orthologous 19 gene clusters (OGC) for accurate taxonomic reassignment resulted in identification of eight 20 distinct and deeply branching clades, further average amino acid identity (AAI) analysis showed 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.07.08.193797; this version posted July 8, 2020.
  • Taxonomy, Identification and Biological Activities of a Novel Isolate of Streptomyces Albus

    Taxonomy, Identification and Biological Activities of a Novel Isolate of Streptomyces Albus

    Kadiri Sunanda Kumari et al. / Journal of Pharmacy Research 2011,4(12),4678-4680 Research Article Available online through ISSN: 0974-6943 www.jpronline.info Taxonomy, Identification and Biological Activities of a Novel Isolate of Streptomyces albus Kadiri Sunanda Kumari* and Vidavalur Siddaiah Department of Organic Chemistry & FDW, Andhra University, Visakhapatnam 530 003, India Received on:20-08-2011; Revised on: 15-09-2011; Accepted on:10-11-2011 ABSTRACT Various isolates of streptomycetes species were isolated from different soil sediments of Visakhapatnam Coast. Preliminary screening by cross-streak method showed that 19 isolates exhibited good antimicrobial activity. Among these 19 isolates tested, isolate MB201 was the most active one and thus was selected for identification. Morphological, cultural, physiological, biochemical characteristics and biological properties as well as enzymatic activities and cell wall composition suggested that the isolate belonged to the genus streptomyces. The 16S ribosomal DNA amplification for phylogenetic study revealed that the isolate was highly related to Streptomyces albus (~99%), so it was designated as Streptomyces albus gangavarams. This new strain produced antimicrobial agents active against Gram +ve and Gram –ve bacteria and fungi. KEY WORDS: Streptomyces, S.albus INTRODUCTION: Numerous classifications were devised to accommodate the increasing number Morphological characterization: of streptomyces species, most of them based on a few subjectively chosen The spore chain morphology of the
  • The Antimicrobial Potential of Streptomyces from Insect Microbiomes

    The Antimicrobial Potential of Streptomyces from Insect Microbiomes

    ARTICLE https://doi.org/10.1038/s41467-019-08438-0 OPEN The antimicrobial potential of Streptomyces from insect microbiomes Marc G. Chevrette 1,2, Caitlin M. Carlson2, Humberto E. Ortega3, Chris Thomas4, Gene E. Ananiev5, Kenneth J. Barns4, Adam J. Book2, Julian Cagnazzo2, Camila Carlos2, Will Flanigan2, Kirk J. Grubbs2, Heidi A. Horn2, F. Michael Hoffmann5, Jonathan L. Klassen6, Jennifer J. Knack 7, Gina R. Lewin8, Bradon R. McDonald 2, Laura Muller2, Weilan G.P. Melo 3, Adrián A. Pinto-Tomás9, Amber Schmitz2, Evelyn Wendt-Pienkowski2, Scott Wildman4, Miao Zhao10, Fan Zhang4, Tim S. Bugni4, David R. Andes10, Monica T. Pupo 3 & Cameron R. Currie2 1234567890():,; Antimicrobial resistance is a global health crisis and few novel antimicrobials have been discovered in recent decades. Natural products, particularly from Streptomyces, are the source of most antimicrobials, yet discovery campaigns focusing on Streptomyces from the soil largely rediscover known compounds. Investigation of understudied and symbiotic sources has seen some success, yet no studies have systematically explored microbiomes for anti- microbials. Here we assess the distinct evolutionary lineages of Streptomyces from insect microbiomes as a source of new antimicrobials through large-scale isolations, bioactivity assays, genomics, metabolomics, and in vivo infection models. Insect-associated Streptomyces inhibit antimicrobial-resistant pathogens more than soil Streptomyces. Genomics and meta- bolomics reveal their diverse biosynthetic capabilities. Further, we describe cyphomycin, a new molecule active against multidrug resistant fungal pathogens. The evolutionary trajectories of Streptomyces from the insect microbiome influence their biosynthetic potential and ability to inhibit resistant pathogens, supporting the promise of this source in augmenting future antimicrobial discovery.