DOCSLIB.ORG

Production and Characterization of Β- Glucanase from Streptomyces Sp

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Production and Characterization of Β- Glucanase from Streptomyces Sp Production and Characterization of β- Glucanase from Streptomyces sp. FINAL TECHNICAL REPORT (Project Reference No: 050/ WSD-BLS/2014/CSTE) Project Period: 01.12.2015 to 30.11.2018 Back To Lab Programme WOMEN SCIENTISTS DIVISION KERALA STATE COUNCIL FOR SCIENCE TECHNOLOGY AND ENVIRONMENT GOVT. OF KERALA LEKSHMI K. EDISON Research Scholar Microbiology Divison KSCSTE-Jawaharlal Nehru Tropical Botanic Garden and Research Institute Palode, Thiruvananthapuram, Kerala, India January 2019 1 AUTHORIZATION The project Production and characterization of β- glucanase from Streptomyces sp. Lekshmi K. Edison, was carried out under the entitled “ Science Technology and Environment,” by Govt. of Kerala. The work was carried“Back out toat Microbiologylab programme” Division, of Women KSCSTE- Scientists Jawaharlal Division, Nehru Kerala Tropical State Botanic Council Gardenfor and Research Institute, Palode under the mentorship of Dr. N. S. Pradeep. The project was initiated on 1st December 2015 with sanction No: 523/2015/KSCSTE dated 14/09/2015, and scheduled completion by 30th November 2018. As per the schedule the field and laboratory works were completed by 30th November 2018 with a financial expenditure of Rs. 16,67,624 lakhs. 2 ACKNOWLEDGMENTS I am deeply indebted to Kerala State Council for Science, Technology and Environment for the financial aid through “Back to Lab program for Women” which helped me to get an independent project with great exposure leading to career development. I am grateful to Dr. K.R. Lekha, Head, Women Scientists Division for the encouragement and support throughout the tenure of the project. I am grateful to Dr. N. S. Pradeep, Scientist mentor, JNTBGRI for giving consent to become the mentor of the Project. I would like to express my gratitude for the encouragement and guidance provided for completion of the work. I am highly obliged to KSCSTE- Jawaharlal Nehru Tropical Botanic Garden and Research Institute for providing me necessary facilities and logistical support for carrying out the a laboratory experiments during the course of the project. I would like to express my deep gratitude to Dr. P. G. Latha, Former Director, for the facilities, support and encouragement provided. I would also like to extend my deep gratitude to Dr. R. Prakash Kumar, Director, for the facilities extended. I wish to record my bountiful thanks to all the scientists, researchers and technical staffs of Microbiology Division. I owe my greatest debt of gratitude to the God Almighty for strengthening me with his love, grace and peace. Lekshmi K. Edison 3 Contents Abstract 5 Abbreviation…………………………………………………………………………………………. ................ 6 Chapter 1. General…………………………………………………………………...... Introduction 7 Chapter 2. Review of Literature …………………………………………………… 14 Chapter 3. Isolation and Screening……………………………………………………. of β-Glucanase Producing Actinomycetes Strains from Western Ghats Regions .. 33 Chapter 4. Identification and Characterization of β-Glucanase Producing Potent Actinomycetes Isolates …… 51 Chapter 5. Isolation, Sequencing and in silico Analysis of β- Glucanase Genes from Streptomyces Strains……………………... ... 67 Chapter 6. Codon Optimization and Overexpression of β-Glucanase genes in E. coli ...................................……………... .. 88 Chapter 7. Purification and Characterization of Overexpressed …………………………………... Recombinant β-Glucanases ............................................................. 102 Summary ………………………….. 112 Achievements ……………………………………………………………. 114 Scope of Future………………………………………………………………………………… Work ..... 121 Major references ………………………………………………………………….... 121 ………………………………………………………………………….. 4 ABSTRACT Beta glucanase (EC 3.2.1.x) are group of glycosyl hydrolase, hydrolyses glycosidic bonds in the - Glucan rich cereals plays numerous beneficial effects. Since it is an immune modulator, it use as a replacement of dietary antibiotics in animal feeds -glucan containinginsoluble beta barley glucan is the molecules. principal β component in breweries, which increases flavour, colour and nutritional properties. Apart from the positive effects, the studies has also been conducted. High β relating with the negative effects. It impairs the energy metabolism in animals such as cattle and poultry. Creates the numerous problems in brewing industry including inefficient filtration, gels and hazes formations which aff -glucanase enzymes -glucan molecules. The currentlyect the existing flavour enzymes and taste are of not beer. capable The marked of the effectample of depolymer β - and its applications- in particular industrial sectors can overcome- the negative effects of β - isation of β glucan.-glucanases. In cereal When β glucan, considering around 70%current of glycosidicdemand andlinkage is β (1→4) and remaining 30% is β- glucanase(1→3). Therefore in brewing the completeindustry andhydrolysis animal offeed this enzyme polymer industry, require we the make synergic a passionate action of attentiondifferent onβ exo-1,4- -glucanase and endo-1,3- - the emerging marketing impact-glucan of β degradation. Streptomyces are the largest genus of phylum actinobacteria, well characterized as the largest reservoirsβ of unhampered naturalβ glucanase, sources projectedof novel tocompounds applicable with in cerealvast industrialβ applications. In this study, we are immensely focused on the isolation of exo- -1,4-glucanase and endo- -1,3-glucanase producing Streptomyces strains by exploring the Western Ghats habitats of Kerala. Then the enzyme encoding genes were isolated and codon optimized forβ obtaining high level expressionβ in E. coli host. The over-expressed enzymes were purified and characterised for the efficient degradation of beta glucan molecule. The codon optimisation of genes showed substantially higher protein expression level as well as enzyme activity than native ones. After nickel affinity purification the optimized exo14 (optexo14) and endo13 (optendo13) protein showed specific activity of 72 U.mg-1 and 65.63 U.mg-1 respectively, which was the highest activities ever reported from Streptomyces spp. Both enzymes showed high thermostability and pH stability. The calculated kinetic parameters of exo14 (Km=7.508, Vmax=58.62) and endo13 - beta glucan. The influence of brewery relevant parameters such as ethanol, ferullic acid, gallic acid and sulphites on(Km=9.221, the activity Vmax=95.51) of individual enzymes exo14, endo13indicating as bothwell ashave cocktail high affinity enzyme towards mix were barley tested. β It revealed, the ethanol, gallic acid and ferullic acid had no significant influence on enzyme activity. The effect of sulphite could be detected around 20% of reduction in relative activity of individual enzymes, cocktail enzyme mix showed more than 90% of activities under varying sulphite concentrations. The results obtained from the present study provide a good understanding for effective industrial producti -glucanase enzymes and suggested the both enzymes, individually as well as cocktail mix, will be the better candidates for the applications in brewing industry onand as animal well as feed utilization industry. of both β Key words: exo- -1,4-glucanase, endo- -1,3-glucanase, codon optimization, over-expression, cocktail mix. β β 5 Abbreviations % Percent G score Glide score °C Degree Celsius GC Guanine cytosine content content µL Microlitre GH Glycosyl hydrolase µmol Micromole GRAVY Grand average of hydropathy AZCL Azurin cross-linked H-bond Hydrogen bond BCC Business Communication Company IMAC Immobilized metal affinity chromatography BLAST Basic Local Alignment Search Tool IPTG Isopropyl β-D-1-thiogalactopyranoside bp Base pair ISP International Streptomyces Project CaCl2 Calcium chloride IUBMB International Union of Biochemistry and Molecular Biology CaCO3 Calcium carbonate IUPAC International Union for Pure and Applied Chemistry CAGR Compound Annual Growth Gate LB Lauria-Bertani CAZy Carbohydrate-Active Enzymes MEGA Molecular Evolutionary Genetics Analysis CBM Carbohydrate binding module min Minute CDD Conserved domain database rRNA Ribosomal RNA MUSCLE Multiple Sequence Comparison by Log- RMSD Root-mean-square deviation Expectation CMC Carboxymethyl cellulose SCA Starch casein agar CM- Carboxymethyl curdlan sdf Structure-data file Curdlan CPD Critical Point Drying SDS Sodium dodecyl sulphate C-score Confidence score sp. Species C-terminal Caboxy-terminal spp. Plural of species D score Docking score TM-score Template modelling Score dbCAN Database for carbohydrate active enzyme UV Ultraviolet radiation annotation dist. H2O Distilled water w/v Weight/volume DNS 3,5-Dinitrosalicylic acid wl Wild dNTPs Deoxyribonucleotide triphosphate x-gal 5-bromo-4-chloro-3-indolyl-β-D- galactopyranoside E.C. Enzyme Commission YEME Yeast extract malt extract EC Electrical conductivity β Beta EDTA Ethylene diamine tetra acetic acid mL Millilitre EI Enzymatic index Fig. Figure ENDO13 Endo-β-1,3-glucanase gene OD Optical density endo13 Endo-β-1,3-glucanase protein BSA Bovine Serum Albumin EtBr Ethidium Bromide kb Kilo base pair EXO14 Exo-β-1,4-glucanase gene KDa Kilo Daltons exo14 Exo-β-1,4-glucanase protein PAGE Polyacrylamide gel electrophoresis G score Glide score cm Centimetre GC Guanine cytosine content CFU.g-1 Colony-forming units per gram content GH Glycosyl hydrolase U.mL-1 Units per millilitre GRAVY Grand average of hydropathy 3D Three dimensional H-bond Hydrogen bond nm Nanomole IMAC Immobilized metal affinity chromatography h Hour 6 Chapter 1 General Introduction 1.1. Industrial Enzymes Biotechnology
Load more

Recommended publications
  • Estimation of Antimicrobial Activities and Fatty Acid Composition Of
    Estimation of antimicrobial activities and fatty acid composition of actinobacteria isolated from water surface of underground lakes from Badzheyskaya and Okhotnichya caves in Siberia Irina V. Voytsekhovskaya1,*, Denis V. Axenov-Gribanov1,2,*, Svetlana A. Murzina3, Svetlana N. Pekkoeva3, Eugeniy S. Protasov1, Stanislav V. Gamaiunov2 and Maxim A. Timofeyev1 1 Irkutsk State University, Irkutsk, Russia 2 Baikal Research Centre, Irkutsk, Russia 3 Institute of Biology of the Karelian Research Centre of the Russian Academy of Sciences, Petrozavodsk, Karelia, Russia * These authors contributed equally to this work. ABSTRACT Extreme and unusual ecosystems such as isolated ancient caves are considered as potential tools for the discovery of novel natural products with biological activities. Acti- nobacteria that inhabit these unusual ecosystems are examined as a promising source for the development of new drugs. In this study we focused on the preliminary estimation of fatty acid composition and antibacterial properties of culturable actinobacteria isolated from water surface of underground lakes located in Badzheyskaya and Okhotnichya caves in Siberia. Here we present isolation of 17 strains of actinobacteria that belong to the Streptomyces, Nocardia and Nocardiopsis genera. Using assays for antibacterial and antifungal activities, we found that a number of strains belonging to the genus Streptomyces isolated from Badzheyskaya cave demonstrated inhibition activity against Submitted 23 May 2018 bacteria and fungi. It was shown that representatives of the genera Nocardia and Accepted 24 September 2018 Nocardiopsis isolated from Okhotnichya cave did not demonstrate any tested antibiotic Published 25 October 2018 properties. However, despite the lack of antimicrobial and fungicidal activity of Corresponding author Nocardia extracts, those strains are specific in terms of their fatty acid spectrum.
    [Show full text]
  • Microbial Factors Associated with the Natural Suppression of Take-All In
    Title Page Microbial factors associated with the natural suppression of take-all in wheat in New Zealand A thesis submitted in partial fulfilment of the requirements for the Degree of Doctor of Philosophy At Lincoln University, Canterbury, New Zealand by Soon Fang Chng Lincoln University 2009 Abstract of a thesis submitted in partial fulfilment of the requirements for the Degree of Doctor of Philosophy Abstract Microbial factors associated with the natural suppression of take- all in wheat in New Zealand by Soon Fang Chng Take-all, caused by the soilborne fungus, Gaeumannomyces graminis var. tritici (Ggt), is an important root disease of wheat that can be reduced by take-all decline (TAD) in successive wheat crops, due to general and/or specific suppression. A study of 112 New Zealand wheat soils in 2003 had shown that Ggt DNA concentrations (analysed using real-time PCR) increased with successive years of wheat crops (1-3 y) and generally reflected take-all severity in subsequent crops. However, some wheat soils with high Ggt DNA concentrations had low take-all, suggesting presence of TAD. This study investigated 26 such soils for presence of TAD and possible suppressive mechanisms, and characterised the microorganisms from wheat roots and rhizosphere using polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE). A preliminary pot trial of 29 soils (including three from ryegrass fields) amended with 12.5% w/w Ggt inoculum, screened their suppressiveness against take-all in a growth chamber. Results indicated that the inoculum level was too high to detect the differences between soils and that the environmental conditions used were unsuitable.
    [Show full text]
  • New Records of Streptomyces and Non Streptomyces Actinomycetes Isolated from Soils Surrounding Sana'a High Mountain
    International Journal of Research in Pharmacy and Biosciences Volume 3, Issue 3, March 2016, PP 19-31 ISSN 2394-5885 (Print) & ISSN 2394-5893 (Online) New Records of Streptomyces and Non Streptomyces Actinomycetes Isolated from Soils Surrounding Sana'a High Mountain Qais Yusuf M. Abdullah1, Maher Ali. Al-Maqtari2, Ola, A A. Al-Awadhi3 Abdullah Y. Al-Mahdi 4 Department of Biology, Microbiology Section Faculty of Science, Sana'a University1, 3, 4 Department of Chemistry, Biochemistry Section, Faculty of Science, Sana'a University2 ABSTRACT Actinomycetes are ubiquitous soil-dwelling saprophytes known to produce secondary metabolites may of which are antibiotic. 20 soil samples were collected from three different sites at high altitude environments surrounding Sana'a city, which ranged from 2300-3000 m above sea level as a prime source of promising native rare actinomycetes. 516 actinomycetes isolates were isolated in pure culture and five selective pretreatment isolation methods. Out of 232 isolates, 26 actinomycetes showing good activity. The identification of 26 selected actinomycetes based on cultural morphology, physiology and biochemical characterization. From the preceding bacteria, thirteen actinomycetes were recorded for the first time from Yemeni soil of these: four non-Streptomyces namely (Intrasporangium sp., Nocardiodes luteus, Sporichthya polymorpha and Streptovirticillium cinnamoneum) and nine Streptomyces namely (S. anulatus, S. celluloflavus, S. cellulosae, S. chromofucus, S. erythrogriseus, S. flavidvirens, S. flavissimus, S. globosus and S. griseoflavus). The results of this study suggested that the soil of high mountains such as Sana'a Mountains could be an interesting source to explore new strains that recorded for the first time in Yemen, Sana'a City.
    [Show full text]
  • Supplementary Information
    Supplementary Information Table S1. Phylum level composition of bacterial communities in eight New Brunswick sediments. Phylum IB * IB% 2B* 2B% 3B * 3B% 4B * 4B% 5B * 5B% 6B * 6B% 7B * 7B% 8B * 8B% Acidobacteria 270 4.0 248 5.3 383 6.4 81 1.0 63 1.4 73 1.4 474 7.5 Actinobacteria 542 8.0 111 2.4 181 3.0 168 2.7 Bacteroidetes 1882 27.7 1133 24.4 1196 20.1 1645 20.9 879 20.1 990 18.7 1450 39.5 2012 32.0 Chlorobi 54 1.2 66 1.2 Planctomycetes 88 1.3 Proteobacteria 2284 33.6 1851 39.9 2468 41.4 4349 55.4 2430 55.4 3020 56.9 1523 41.5 2510 39.9 Verrucomicrobia 307 4.5 133 3.6 65 1.0 Unclassified Bacteria 1367 20.1 1205 26.0 1581 26.5 1474 18.8 824 18.8 1058 19.9 487 13.3 926 14.7 “Rare Phyla” 51 0.8 95 2.0 148 2.5 223 2.8 133 3.0 99 1.9 79 2.2 128 2.0 Total 6791 100.0 4643 100.0 5957 100.0 7856 100.0 4383 100.0 5306 100.0 3672 100.0 6283 100.0 * Denotes number of sequences. Table S2. Number of sequences represented in the “Rare Phyla” group presented in Table S1. Phylum IB 2B 3B 4B 5B 6B 7B 8B Acidobacteria 5 Actinobacteria 62 39 33 13 Armatimonadetes 1 9 Chlorobi 13 2 5 5 Chloroflexi 3 4 4 12 27 8 6 20 Fusobacteria 3 1 1 1 Gemmatimonadetes 9 1 1 9 Lentisphaerae 4 2 2 2 12 4 Nitrospira 5 1 22 2 36 Planctomycetes 37 55 52 18 21 9 28 Spirochaetes 2 1 Verrucomicrobia 46 51 73 33 26 Mar.
    [Show full text]
  • Diversity of Antibiotic-Active Bacteria Associated with the Brown Alga Laminaria Saccharina from the Baltic Sea
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by OceanRep Mar Biotechnol (2009) 11:287–300 DOI 10.1007/s10126-008-9143-4 ORIGINAL ARTICLE Diversity of Antibiotic-Active Bacteria Associated with the Brown Alga Laminaria saccharina from the Baltic Sea Jutta Wiese & Vera Thiel & Kerstin Nagel & Tim Staufenberger & Johannes F. Imhoff Received: 28 January 2008 /Accepted: 1 September 2008 /Published online: 15 October 2008 # Springer Science + Business Media, LLC 2008 Abstract Bacteria associated with the marine macroalga (Bartsch et al. 2008). Epiphytic bacteria have been studied Laminaria saccharina, collected from the Kiel Fjord (Baltic by microscopic methods (Corre and Prieur 1990) and by Sea, Germany), were isolated and tested for antimicrobial genetic and cultivation approaches. Bacterial cell numbers activity. From a total of 210 isolates, 103 strains inhibited of up to 107 colony-forming units (CFU) per centimeter the growth of at least one microorganism from the test squared were reported for Laminaria digitata from the panel including Gram-negative and Gram-positive bacteria coast of Brittany (France) and Laminaria pallida on the as well as a yeast. Most common profiles were the Bengal upwelling region of southern Africa (Corre and inhibition of Bacillus subtilis only (30%), B. subtilis and Prieur 1990; Mazure and Field 1980). However, the Staphylococcus lentus (25%), and B. subtilis, S. lentus, and interactions between members of the epiphytic and endo- Candida albicans (11%). In summary, the antibiotic-active phytic communities and the relationships between these isolates covered 15 different activity patterns suggesting communities and Laminaria spp.
    [Show full text]
  • F Ull L Ength Original R Esearch P Aper
    Copyright © 2013 By IYPF All rights reserved Open Access Contents Int. J. Drug Dev. & Res . | July-September 2013 | Vol. 5 | Issue 3 | ISSN 0975-9344 | www.ijddr.in Biological activities of Streptomyces species SRDP-07 isolated from soil of Thirthahalli, Karnataka, India Prashith Kekuda T. R 1,2* Abstract: Western Ghats are one of the biodiversity hotspots in the w orld. The present study was undertaken to investigate biological activities viz., antibacterial, 1 Dileep N insecticidal, anthelmintic and antioxidant activities of a Streptomyces species SRDP-07 isolated from a soil sample collected in Thirthahalli, Karnataka (state ), 1 FullLength Syed Junaid India. Isolation of actinomycetes was carried out by plating the serially diluted soil sample on Starch casein agar. Primary screening of the actinomycete isolates for 1 antibacterial activity was done by Cross streak method. Isolate SRDP-07, showing Rakesh. K. N prom inent inhibition of test bacteria in cross streak technique, was selected for further characterization and for determining biological activities. The isolate was 2 Sunita C Mesta grown in starch casein broth for 10 days and the culture filtrate was extracted with ethyl ace tate. Antibacterial activity of ethyl acetate extract was tested against a panel of 6 bacteria by Agar well diffusion assay. Insecticidal activity of the extract Onkarappa R 2 was evaluated in terms of larvicidal effect against 2 nd instar larvae of Aedes aegypti . Anthel mintic activity of the extract was performed by determining time 1 Department of Microbiology, taken for paralysis and death of adult Indian earthworms. Antioxidant activity of Page 268 Page 268 S.R.N.M.N College of Applied the extract was determined by DPPH free radical scavenging and Ferric reducing Sciences, NES Campus, Balraj assay.
    [Show full text]
  • African Journal of Biotechnology
    OPEN ACCESS African Journal of Biotechnology 20 September 2010 ISSN 1684-5315 DOI: 10.5897/AJB www.academicjournals.org About AJB The African Journal of Biotechnology (AJB) is a peer reviewed journal which commenced publication in 2002. AJB publishes articles from all areas of biotechnology including medical and pharmaceutical biotechnology, molecular diagnostics, applied biochemistry, industrial microbiology, molecular biology, bioinformatics, genomics and proteomics, transcriptomics and genome editing, food and agricultural technologies, and metabolic engineering. Manuscripts on economic and ethical issues relating to biotechnology research are also considered. Indexing CAB Abstracts, CABI’s Global Health Database, Chemical Abstracts (CAS Source Index) Dimensions Database, Google Scholar, Matrix of Information for The Analysis of Journals (MIAR), Microsoft Academic, Research Gate Open Access Policy Open Access is a publication model that enables the dissemination of research articles to the global community without restriction through the internet. All articles published under open access can be accessed by anyone with internet connection. The African Journals of Biotechnology is an Open Access journal. Abstracts and full texts of all articles published in this journal are freely accessible to everyone immediately after publication without any form of restriction. Article License All articles published by African Journal of Biotechnology are licensed under the Creative Commons Attribution 4.0 International License. This permits anyone
    [Show full text]
  • Phylogenetic Study of the Species Within the Family Streptomycetaceae
    Antonie van Leeuwenhoek DOI 10.1007/s10482-011-9656-0 ORIGINAL PAPER Phylogenetic study of the species within the family Streptomycetaceae D. P. Labeda • M. Goodfellow • R. Brown • A. C. Ward • B. Lanoot • M. Vanncanneyt • J. Swings • S.-B. Kim • Z. Liu • J. Chun • T. Tamura • A. Oguchi • T. Kikuchi • H. Kikuchi • T. Nishii • K. Tsuji • Y. Yamaguchi • A. Tase • M. Takahashi • T. Sakane • K. I. Suzuki • K. Hatano Received: 7 September 2011 / Accepted: 7 October 2011 Ó Springer Science+Business Media B.V. (outside the USA) 2011 Abstract Species of the genus Streptomyces, which any other microbial genus, resulting from academic constitute the vast majority of taxa within the family and industrial activities. The methods used for char- Streptomycetaceae, are a predominant component of acterization have evolved through several phases over the microbial population in soils throughout the world the years from those based largely on morphological and have been the subject of extensive isolation and observations, to subsequent classifications based on screening efforts over the years because they are a numerical taxonomic analyses of standardized sets of major source of commercially and medically impor- phenotypic characters and, most recently, to the use of tant secondary metabolites. Taxonomic characteriza- molecular phylogenetic analyses of gene sequences. tion of Streptomyces strains has been a challenge due The present phylogenetic study examines almost all to the large number of described species, greater than described species (615 taxa) within the family Strep- tomycetaceae based on 16S rRNA gene sequences Electronic supplementary material The online version and illustrates the species diversity within this family, of this article (doi:10.1007/s10482-011-9656-0) contains which is observed to contain 130 statistically supplementary material, which is available to authorized users.
    [Show full text]
  • Molecular and Biological Studies of Streptomyces Sp. Producing Antibacterial Agents Against Some Pathogenic Bacteria
    International Journal of Pharmaceutical and Phytopharmacological Research (eIJPPR) | February 2021 | Volume 11| Issue 1 | Page 125-133 Afra M. Baghdadi, Molecular and Biological Studies of Streptomyces sp. Producing Antibacterial Agents against Some Pathogenic Bacteria Molecular and Biological Studies of Streptomyces sp. Producing Antibacterial Agents against Some Pathogenic Bacteria Afra M. Baghdadi*, Naylah M. Alblady Department of Biology Sciences, Faculty of Science, University of Jeddah, Jeddah, Saudi Arabia. ABSTRACT The genus Streptomyces, known as prolific antibiotic products and many other bioactive secondary metabolites, is a unique subgroup of actinomycetes bacteria. This study aims to isolate Streptomyces strains from plant rhizosphere that has antibacterial activity. Soil samples were collected from Saudi Arabia. About 39 isolates of Streptomyces sp. were obtained from 12 soil samples, using serial dilution and plating techniques on the solid starch nitrate medium. The Streptomyces strains were cross streaked against some human pathogen bacteria. Against various pathogenic bacteria, only 16 out of 39 isolates demonstrated strong antimicrobial activity. The most active strain Streptomyces NM38 was chosen for secondary screening and identification. The molecular identification was confirmed using 16S rRNA as Streptomyces globosus, the phylogenetic tree was built; the sequence was presented to the GenBank under the accession number: MN538259.1. Key Words: Antibacterial activity, Streptomyces, Pathogenic bacteria, Identification. eIJPPR 2021; 11(1):125-133 HOW TO CITE THIS ARTICLE: Afra M. Baghdadi, Naylah M. Alblady (2021). Molecular and Biological Studies of Streptomyces sp. Producing Antibacterial Agents against Some Pathogenic Bacteria , International Journal of Pharmaceutical and Phytopharmacological Research, 11(1), pp.125-133. “ ” INTRODUCTION Microbial diseases are growing year after year and have metabolites.
    [Show full text]
  • Description of Unrecorded Bacterial Species Belonging to the Phylum Actinobacteria in Korea
    Journal of Species Research 10(1):23­45, 2021 Description of unrecorded bacterial species belonging to the phylum Actinobacteria in Korea Mi­Sun Kim1, Seung­Bum Kim2, Chang­Jun Cha3, Wan­Taek Im4, Won­Yong Kim5, Myung­Kyum Kim6, Che­Ok Jeon7, Hana Yi8, Jung­Hoon Yoon9, Hyung­Rak Kim10 and Chi­Nam Seong1,* 1Department of Biology, Sunchon National University, Suncheon 57922, Republic of Korea 2Department of Microbiology, Chungnam National University, Daejeon 34134, Republic of Korea 3Department of Biotechnology, Chung-Ang University, Anseong 17546, Republic of Korea 4Department of Biotechnology, Hankyong National University, Anseong 17579, Republic of Korea 5Department of Microbiology, College of Medicine, Chung-Ang University, Seoul 06974, Republic of Korea 6Department of Bio & Environmental Technology, Division of Environmental & Life Science, College of Natural Science, Seoul Women’s University, Seoul 01797, Republic of Korea 7Department of Life Science, Chung-Ang University, Seoul 06974, Republic of Korea 8School of Biosystem and Biomedical Science, Korea University, Seoul 02841, Republic of Korea 9Department of Food Science and Biotechnology, Sungkyunkwan University, Suwon 16419, Republic of Korea 10Department of Laboratory Medicine, Saint Garlo Medical Center, Suncheon 57931, Republic of Korea *Correspondent: [email protected] For the collection of indigenous prokaryotic species in Korea, 77 strains within the phylum Actinobacteria were isolated from various environmental samples, fermented foods, animals and clinical specimens in 2019. Each strain showed high 16S rRNA gene sequence similarity (>98.8%) and formed a robust phylogenetic clade with actinobacterial species that were already defined and validated with nomenclature. There is no official description of these 77 bacterial species in Korea.
    [Show full text]
  • High Diversity and Suggested Endemicity of Culturable Actinobacteria in an Extremely Oligotrophic Desert Oasis
    High diversity and suggested endemicity of culturable Actinobacteria in an extremely oligotrophic desert oasis Hector Fernando Arocha-Garza1, Ricardo Canales-Del Castillo2, Luis E. Eguiarte3, Valeria Souza3 and Susana De la Torre-Zavala1 1 Facultad de Ciencias Biológicas, Instituto de Biotecnología, Universidad Autónoma de Nuevo León, San Nicolás de los Garza, Nuevo León, Mexico 2 Facultad de Ciencias Biológicas, Laboratorio de Biología de la Conservación, Universidad Autónoma de Nuevo León, San Nicolás de los Garza, Nuevo León, Mexico 3 Departamento de Ecología Evolutiva, Instituto de Ecología, Universidad Nacional Autónoma de México, Mexico City, Mexico ABSTRACT The phylum Actinobacteria constitutes one of the largest and anciently divergent phyla within the Bacteria domain. Actinobacterial diversity has been thoroughly researched in various environments due to its unique biotechnological potential. Such studies have focused mostly on soil communities, but more recently marine and extreme environments have also been explored, finding rare taxa and demonstrating dispersal limitation and biogeographic patterns for Streptomyces. To test the distribution of Actinobacteria populations on a small scale, we chose the extremely oligotrophic and biodiverse Cuatro Cienegas Basin (CCB), an endangered oasis in the Chihuahuan desert to assess the diversity and uniqueness of Actinobacteria in the Churince System with a culture-dependent approach over a period of three years, using nine selective media. The 16S rDNA of putative Actinobacteria were sequenced using both bacteria universal and phylum-specific primer pairs. Phylogenetic reconstructions were performed to analyze OTUs clustering and taxonomic identification of the isolates in an evolutionary context, using validated type species of Streptomyces from previously phylogenies as a reference.
    [Show full text]
  • Diversity of Actinobacteria Isolated from Date Palms Rhizosphere and Saline Environments: Isolation, Identification and Biological Activity Evaluation
    microorganisms Article Diversity of Actinobacteria Isolated from Date Palms Rhizosphere and Saline Environments: Isolation, Identification and Biological Activity Evaluation Omar Messaoudi 1,2,3,* , Joachim Wink 3 and Mourad Bendahou 1 1 Laboratory of Applied Microbiology in Food, Biomedical and Environment, Abou Bekr Belkaïd University, 13000 Tlemcen, Algeria; [email protected] 2 Department of Biology, Faculty of Science, University of Amar Telidji, 03000 Laghouat, Algeria 3 Microbial Strain Collection, Helmholtz Centre for Infection Research GmbH (HZI), Inhoffenstrasse 7, 38124 Braunschweig, Germany; [email protected] * Correspondence: [email protected] or [email protected]; Tel.: +213-557084015 Received: 1 October 2020; Accepted: 2 November 2020; Published: 25 November 2020 Abstract: The diversity of cultural Actinobacteria in two types of Algerian Sahara environments, including saline environments and date palms rhizosphere, was investigated. In this study, a total of 40 strains of actinomycetes was isolated from different soil samples, using a rehydration and centrifugation method. Molecular identification, based on 16S rRNA gene sequence analysis, revealed that these isolates were affiliated to six clusters corresponding to eight genera, including Streptomyces, Nocardiopsis, Saccharopolyspora, Actinomadura, Actinocorallia, Micromonospora, Couchioplanes, and Planomonospora. A taxonomic analysis, based on the morphological, physiological, biochemical, and molecular investigation, of selected strains, which
    [Show full text]