DOCSLIB.ORG

Insight Into the Molecular Mechanisms for Microcystin Biodegradation in Lake Erie and Lake Taihu

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Insight Into the Molecular Mechanisms for Microcystin Biodegradation in Lake Erie and Lake Taihu University of Tennessee, Knoxville TRACE: Tennessee Research and Creative Exchange Microbiology Publications and Other Works Microbiology 2019 Insight Into the Molecular Mechanisms for Microcystin Biodegradation in Lake Erie and Lake Taihu Lauren E. Krausfeldt University of Tennessee, Knoxville Morgan M. Steffen James Madison University, Harrisonburg Robert M. McKay Great Lakes Institute for Environmental Research, University of Windsor, Windsor, ON, Canada George S. Bullerjahn Bowling Green State University, Bowling Green Gregory L. Boyer College of Environmental Science and Forestry, State University of New York, Syracuse, See next page for additional authors Follow this and additional works at: https://trace.tennessee.edu/utk_micrpubs Recommended Citation Krausfeldt, Lauren E.; Steffen, Morgan M.; McKay, Robert M.; Bullerjahn, George S.; Boyer, Gregory L.; and Wilhelm, Steven W., "Insight Into the Molecular Mechanisms for Microcystin Biodegradation in Lake Erie and Lake Taihu" (2019). Microbiology Publications and Other Works. https://trace.tennessee.edu/utk_micrpubs/110 This Article is brought to you for free and open access by the Microbiology at TRACE: Tennessee Research and Creative Exchange. It has been accepted for inclusion in Microbiology Publications and Other Works by an authorized administrator of TRACE: Tennessee Research and Creative Exchange. For more information, please contact [email protected]. Authors Lauren E. Krausfeldt, Morgan M. Steffen, Robert M. McKay, George S. Bullerjahn, Gregory L. Boyer, and Steven W. Wilhelm This article is available at TRACE: Tennessee Research and Creative Exchange: https://trace.tennessee.edu/ utk_micrpubs/110 fmicb-10-02741 December 7, 2019 Time: 11:11 # 1 ORIGINAL RESEARCH published: 10 December 2019 doi: 10.3389/fmicb.2019.02741 Insight Into the Molecular Mechanisms for Microcystin Biodegradation in Lake Erie and Lake Taihu Lauren E. Krausfeldt1, Morgan M. Steffen2, Robert M. McKay3, George S. Bullerjahn4, Gregory L. Boyer5 and Steven W. Wilhelm1* 1 Department of Microbiology, The University of Tennessee, Knoxville, Knoxville, TN, United States, 2 Department of Biology, James Madison University, Harrisonburg, VA, United States, 3 Great Lakes Institute for Environmental Research, University of Windsor, Windsor, ON, Canada, 4 Department of Biological Sciences, Bowling Green State University, Bowling Green, OH, United States, 5 Department of Chemistry, College of Environmental Science and Forestry, State University of New York, Syracuse, NY, United States Microcystins are potent hepatotoxins that are frequently detected in fresh water lakes plagued by toxic cyanobacteria. Microbial biodegradation has been referred to as the most important avenue for removal of microcystin from aquatic environments. The Edited by: biochemical pathway most commonly associated with the degradation of microcystin Pia H. Moisander, University of Massachusetts is encoded by the mlrABCD (mlr) cassette. The ecological significance of this pathway Dartmouth, United States remains unclear as no studies have examined the expression of these genes in natural Reviewed by: environments. Six metatranscriptomes were generated from microcystin-producing Fei Yang, Central South University, China Microcystis blooms and analyzed to assess the activity of this pathway in environmental Rachel Susan Poretsky, samples. Seventy-eight samples were collected from Lake Erie, United States/Canada University of Illinois at Chicago, and Lake Tai (Taihu), China, and screened for the presence of mlr gene transcripts. United States Read mapping to the mlr cassette indicated transcripts for these genes were absent, *Correspondence: Steven W. Wilhelm with only 77 of the collective 3.7 billion reads mapping to any part of the mlr cassette. [email protected] Analysis of the assembled metatranscriptomes supported this, with only distantly related sequences identified as mlrABC-like. These observations were made despite Specialty section: This article was submitted to the presence of microcystin and over 500,000 reads mapping to the mcy cassette for Aquatic Microbiology, microcystin production. Glutathione S-transferases and alkaline proteases have been a section of the journal Frontiers in Microbiology previously hypothesized to be alternative pathways for microcystin biodegradation, and Received: 02 July 2019 expression of these genes was detected across space and time in both lakes. While the Accepted: 11 November 2019 activity of these alternative pathways needs to be experimentally confirmed, they may Published: 10 December 2019 be individually or collectively more important than mlr genes in the natural environment. Citation: Krausfeldt LE, Steffen MM, Importantly, the lack of mlr expression could indicate microcystin biodegradation was McKay RM, Bullerjahn GS, Boyer GL not occurring in the analyzed samples. This study raises interesting questions about the and Wilhelm SW (2019) Insight Into ubiquity, specificity and locality of microcystin biodegradation, and highlights the need the Molecular Mechanisms for Microcystin Biodegradation for the characterization of relevant mechanisms in natural communities to understand in Lake Erie and Lake Taihu. the fate of microcystin in the environment and risk to public health. Front. Microbiol. 10:2741. doi: 10.3389/fmicb.2019.02741 Keywords: biodegradation, cyanotoxins, cyanobacteria, harmful algal blooms, RNA-sequencing Frontiers in Microbiology | www.frontiersin.org 1 December 2019 | Volume 10 | Article 2741 fmicb-10-02741 December 7, 2019 Time: 11:11 # 2 Krausfeldt et al. Microcystin Biodegradation in Large Lakes INTRODUCTION the most important route for the disappearance of MCs in nature (Christoffersen et al., 2002; Holst et al., 2003; Chen et al., 2008; Microcystins (MCs) are the one of the most frequently detected Grützmacher et al., 2009; Bukowska et al., 2018). cyanotoxins within freshwater harmful cyanobacterial blooms The reported genetic pathway involved in MC biodegradation (cyanoHABs). With cyanoHABS on the rise around the world, involves four enzymes encoded by mlrA, mlrB, mlrC, and mlrD understanding the fate of MCs in the environment has assumed (Bourne et al., 1996; Bourne et al., 2001). This pathway has greater importance as they pose both ecological and public health been primarily associated with the biodegradation of MC-LR, risks. MCs can be potent protein phosphatase 1 and 2A inhibitors a prevalent congener in nature (Carmichael and Boyer, 2016). as well as potential tumor promoters. Consumption of MCs can MlrA, termed microcystinase, is a metalloprotease linearizing result in acute hepatocytosis, cancer and various gastrointestinal the MC-LR at the Arg-ADDA bond. It is commonly used as problems (Carmichael, 2001; Zurawell et al., 2005; Carmichael marker gene and considered to catalyze the most important and Boyer, 2016). Conventional municipal water treatments are step in biodegradation of MCs (Dziga et al., 2012; Dexter effective at removing or inactivating cyanobacterial cells, but et al., 2018; Lezcano et al., 2018). MlrB is a serine protease the methods risk the release of MCs from the cell. MCs are that drives the hydrolysis of the linearized MC-LR at the Ala- not removed by flocculation processes and require secondary Leu bond, forming a tetrapeptide. The tetrapeptide is then treatments that include the use of activated carbon, which is subject to further degradation by a second metalloprotease, effective but costly (He et al., 2016). Indeed, liver cancer and MlrC, and it was demonstrated that MlrC can also cleave colorectal cancer has been linked to nearby bodies of water the linearized MC at the -Adda group (Dziga et al., 2012). plagued with toxic cyanobacteria in the United States, Serbia MlrD is a putative transporter, but whether MC biodegradation and China (Carmichael, 2001; Hernández et al., 2009; Svircev occurs intracellularly or extracellularly remains inconclusive et al., 2009). Cases of mortality and morbidity due to cyanotoxins (Bourne et al., 2006). have been reported since the 1800s involving birds, livestock, This pathway has been extensively studied in culture dogs, fish, and even humans (Chorus and Bartram, 1999; Wood, (Supplementary Table S1, Bourne et al., 2006; Dziga et al., 2012; 2016). The number of cases has risen gradually over time and Maseda et al., 2012; Dziga et al., 2016; Dexter et al., 2018;as it is predicted that the number of individuals affected is greatly examples), although not all MC degrading isolates have tested underestimated (Wood, 2016). positive for these genes. Further, the environmental relevance Microcystins are chemically stable and resistant to many and distribution of the mlr cassette in the environment remains abiotic factors, which has led to numerous studies questioning unclear. The mlrA gene has been detected in DNA isolated their fate in the environment (Harada and Tsuji, 1998). Whereas from microbial communities with MC degradation capabilities their disappearance has been attributed to some abiotic effects from freshwater samples (Mou et al., 2013; Kansole and Lin, including dilution, adsorption, and photodegradation (Harada 2016; Lezcano et al., 2018), but more commonly environments and Tsuji, 1998; Corbel et al., 2014), they are generally where MC biodegradation has been tested are not screened considered to be resistant to abiotic factors. MCs are also for use of this pathway. More often, the mlrA gene has been resistant to the activity of common peptidases (Tsuji et al., detected in enriched samples such as
Load more

Recommended publications
  • The Fate of Microcystins in the Environment and Challenges for Monitoring
    Toxins 2014, 6, 3354-3387; doi:10.3390/toxins6123354 OPEN ACCESS toxins ISSN 2072-6651 www.mdpi.com/journal/toxins Review The Fate of Microcystins in the Environment and Challenges for Monitoring Justine R. Schmidt 1,*, Steven W. Wilhelm 2 and Gregory L. Boyer 1 1 Department of Chemistry, College of Environmental Science and Forestry, State University of New York, Syracuse, NY 13210, USA; E-Mail: [email protected] 2 Department of Microbiology, University of Tennessee, Knoxville, TN 37996-0845, USA; E-Mail: [email protected] * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +1-315-470-6844; Fax: +1-315-470-6856. External Editor: Lesley V. D'Anglada Received: 1 November 2014; in revised form: 29 November 2014 / Accepted: 5 December 2014 / Published: 12 December 2014 Abstract: Microcystins are secondary metabolites produced by cyanobacteria that act as hepatotoxins in higher organisms. These toxins can be altered through abiotic processes, such as photodegradation and adsorption, as well as through biological processes via metabolism and bacterial degradation. Some species of bacteria can degrade microcystins, and many other organisms metabolize microcystins into a series of conjugated products. There are toxicokinetic models used to examine microcystin uptake and elimination, which can be difficult to compare due to differences in compartmentalization and speciation. Metabolites of microcystins are formed as a detoxification mechanism, and little is known about how quickly these metabolites are formed. In summary, microcystins can undergo abiotic and biotic processes that alter the toxicity and structure of the microcystin molecule. The environmental impact and toxicity of these alterations and the metabolism of microcystins remains uncertain, making it difficult to establish guidelines for human health.
    [Show full text]
  • Genome Diversification in Globally Distributed Novel Marine
    bioRxiv preprint doi: https://doi.org/10.1101/814418; this version posted October 22, 2019. 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 Genome diversification in globally distributed novel 2 marine Proteobacteria is linked to environmental adaptation 3 4 Authors: Zhichao Zhou1, Patricia Q. Tran1, 2, Kristopher Kieft1, Karthik Anantharaman1* 5 6 7 Affiliations: 8 1Department of Bacteriology, University of Wisconsin–Madison, Madison, WI, 53706, USA 9 2Department of Integrative Biology, University of Wisconsin–Madison, Madison, WI 53706, 10 USA 11 12 13 *Corresponding author 14 Email: [email protected] 15 Address: 4550 Microbial Sciences Building, 1550 Linden Dr., Madison, WI, 53706 16 17 Abstract 18 Proteobacteria constitute the most diverse and abundant group of microbes on Earth. In 19 productive marine environments like deep-sea hydrothermal systems, Proteobacteria have been 20 implicated in autotrophy coupled to sulfur, methane, and hydrogen oxidation, sulfate reduction, 21 and denitrification. Beyond chemoautotrophy, little is known about the ecological significance 22 of novel Proteobacteria that are globally distributed and active in hydrothermal systems. Here 23 we apply multi-omics to characterize 51 metagenome-assembled genomes from three 24 hydrothermal vent plumes in the Pacific and Atlantic Oceans that are affiliated with nine novel 25 Proteobacteria lineages. Metabolic analyses revealed these organisms to contain a diverse 26 functional repertoire including chemolithotrophic ability to utilize sulfur and C1 compounds, 27 and chemoorganotrophic ability to utilize environment-derived fatty acids, aromatics, 28 carbohydrates, and peptides.
    [Show full text]
  • <I>MICROCYSTIS</I> BLOOMS
    University of Tennessee, Knoxville TRACE: Tennessee Research and Creative Exchange Doctoral Dissertations Graduate School 8-2018 MOLECULAR CHARACTERIZATION OF FACTORS CONSTRAINING THE SUCCESS AND TOXICITY OF MICROCYSTIS BLOOMS Lauren Elisabeth Krausfeldt University of Tennessee, [email protected] Follow this and additional works at: https://trace.tennessee.edu/utk_graddiss Recommended Citation Krausfeldt, Lauren Elisabeth, "MOLECULAR CHARACTERIZATION OF FACTORS CONSTRAINING THE SUCCESS AND TOXICITY OF MICROCYSTIS BLOOMS. " PhD diss., University of Tennessee, 2018. https://trace.tennessee.edu/utk_graddiss/5030 This Dissertation is brought to you for free and open access by the Graduate School at TRACE: Tennessee Research and Creative Exchange. It has been accepted for inclusion in Doctoral Dissertations by an authorized administrator of TRACE: Tennessee Research and Creative Exchange. For more information, please contact [email protected]. To the Graduate Council: I am submitting herewith a dissertation written by Lauren Elisabeth Krausfeldt entitled "MOLECULAR CHARACTERIZATION OF FACTORS CONSTRAINING THE SUCCESS AND TOXICITY OF MICROCYSTIS BLOOMS." I have examined the final electronic copy of this dissertation for form and content and recommend that it be accepted in partial fulfillment of the requirements for the degree of Doctor of Philosophy, with a major in Microbiology. Steven W. Wilhelm, Major Professor We have read this dissertation and recommend its acceptance: Alison Buchan, Shawn R. Campagna, Karen G. Lloyd Accepted for the Council: Dixie L. Thompson Vice Provost and Dean of the Graduate School (Original signatures are on file with official studentecor r ds.) MOLECULAR CHARACTERIZATION OF FACTORS CONSTRAINING THE SUCCESS AND TOXICITY OF MICROCYSTIS BLOOMS A Dissertation Presented for the Doctor of Philosophy Degree The University of Tennessee, Knoxville Lauren Elisabeth Krausfeldt August 2018 Copyright © 2018 by Lauren E.
    [Show full text]
  • Extracellular Enzymes in Aquatic Environments
    University of Tennessee, Knoxville TRACE: Tennessee Research and Creative Exchange Masters Theses Graduate School 8-2020 Extracellular Enzymes in Aquatic Environments: Possible Role of Non-Specific eptidasesP in Microcystin Degradation and Effects of Assay Protocol on Calculated Activities Christopher Cook University of Tennessee Follow this and additional works at: https://trace.tennessee.edu/utk_gradthes Recommended Citation Cook, Christopher, "Extracellular Enzymes in Aquatic Environments: Possible Role of Non-Specific Peptidases in Microcystin Degradation and Effects of Assay Protocol on Calculated Activities. " Master's Thesis, University of Tennessee, 2020. https://trace.tennessee.edu/utk_gradthes/6073 This Thesis is brought to you for free and open access by the Graduate School at TRACE: Tennessee Research and Creative Exchange. It has been accepted for inclusion in Masters Theses by an authorized administrator of TRACE: Tennessee Research and Creative Exchange. For more information, please contact [email protected]. To the Graduate Council: I am submitting herewith a thesis written by Christopher Cook entitled "Extracellular Enzymes in Aquatic Environments: Possible Role of Non-Specific eptidasesP in Microcystin Degradation and Effects of Assay Protocol on Calculated Activities." I have examined the final electronic copy of this thesis for form and content and recommend that it be accepted in partial fulfillment of the requirements for the degree of Master of Science, with a major in Geology. Andrew Steen, Major Professor We have
    [Show full text]
  • In Silico Proteomic Analysis Provides Insights Into Phylogenomics and Plant Biomass Deconstruction Potentials of the Tremelalles
    fbioe-08-00226 April 2, 2020 Time: 17:58 # 1 ORIGINAL RESEARCH published: 03 April 2020 doi: 10.3389/fbioe.2020.00226 In silico Proteomic Analysis Provides Insights Into Phylogenomics and Plant Biomass Deconstruction Potentials of the Tremelalles Habibu Aliyu1*†, Olga Gorte1†, Xinhai Zhou1,2, Anke Neumann1 and Katrin Ochsenreither1* 1 Institute of Process Engineering in Life Science 2: Technical Biology, Karlsruhe Institute of Technology, Karlsruhe, Germany, 2 State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing, China Edited by: Basidiomycetes populate a wide range of ecological niches but unlike ascomycetes, Joao Carlos Setubal, their capabilities to decay plant polymers and their potential for biotechnological University of São Paulo, Brazil approaches receive less attention. Particularly, identification and isolation of CAZymes Reviewed by: Marco Antonio Seiki Kadowaki, is of biotechnological relevance and has the potential to improve the cache of currently Université libre de Bruxelles, Belgium available commercial enzyme cocktails toward enhanced plant biomass utilization. The Renato Graciano De Paula, order Tremellales comprises phylogenetically diverse fungi living as human pathogens, Federal University of Espírito Santo, Brazil mycoparasites, saprophytes or associated with insects. Here, we have employed Gabriel Paes, comparative genomics approaches to highlight the phylogenomic relationships among Fractionnation of AgroResources and Environment (INRA), France thirty-five Tremellales and to identify putative enzymes of biotechnological interest *Correspondence: encoded on their genomes. Evaluation of the predicted proteomes of the thirty-five Habibu Aliyu Tremellales revealed 6,918 putative carbohydrate-active enzymes (CAZYmes) and 7,066 [email protected]; peptidases. Two soil isolates, Saitozyma podzolica DSM 27192 and Cryptococcus sp.
    [Show full text]
  • Elucidating the Regulatory Functions of Mlra Originated from Novosphingobium Sp
    ntal & A me na n ly o t ir ic Li et al., J Environ Anal Toxicol 2018, 8:2 v a n l T E o Journal of f x DOI: 10.4172/2161-0525.1000556 o i l c o a n l o r g u y o J Environmental & Analytical Toxicology ISSN: 2161-0525 ResearchResearch Article Article OpenOpen Access Access Elucidating the Regulatory Functions of MlrA Originated from Novosphingobium sp. THN1 in Microcystin-LR Degradation Jieming Li, Ruiping Wang and Ji Li* College of Resources and Environmental Sciences, China Agricultural University, Beijing 100193, PR China Abstract Microcystin-LR (MC-LR), produced by harmful cyanobacteria, seriously endangers animals and humans. Biodegradation appears as the major pathway for natural MC-LR attenuation. To elucidate the regulatory function of mlrA gene of Novosphingobium sp. THN1 (i.e., THN1-mlrA gene) in MC-LR biodegradation, this study constructed a recombinant bacterium and succeeded in heterlogously expressing the MlrA of THN1 strain (i.e., THN1-MlrA enzyme). The recombinant MlrA exhibited the activity for smoothly degrading 20 μg mL-1 of MC-LR at an average rate of 0.16 μg mL-1 h-1 within 80 h. Mass spectrum analysis confirmed that recombinant MlrA hydrolyzed cyclic MC-LR by cleaving the peptide bond between Adda and arginine residue and generated linearized MC-LR as primary intermediate. Such linearization for MC-LR catalyzed by THN1-MlrA enzyme was particularly important during MC-LR biodegradation process, because it opened the highly-stable cyclic structure of MC-LR and caused substantial detoxification. These findings for the first time manifested thatmlrA gene homolog of Novosphingobium genus conserved its original catalytic function as described elsewhere.
    [Show full text]
  • Synthese Et Evaluation Biologique De Derives De L'aminobenzosuberone
    Synthese et evaluation biologique de derives de l'aminobenzosuberone, inhibiteurs puissants et selectifs de l'aminopeptidase N ou CD13 Sarah Alavi To cite this version: Sarah Alavi. Synthese et evaluation biologique de derives de l'aminobenzosuberone, inhibiteurs puissants et selectifs de l'aminopeptidase N ou CD13. Chimie organique. Universit´ede Haute Alsace - Mulhouse, 2013. Fran¸cais. <NNT : 2013MULH4071>. <tel-01223670> HAL Id: tel-01223670 https://tel.archives-ouvertes.fr/tel-01223670 Submitted on 3 Nov 2015 HAL is a multi-disciplinary open access L'archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destin´eeau d´ep^otet `ala diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publi´esou non, lished or not. The documents may come from ´emanant des ´etablissements d'enseignement et de teaching and research institutions in France or recherche fran¸caisou ´etrangers,des laboratoires abroad, or from public or private research centers. publics ou priv´es. THESE En vue de l’obtention du Présentée et soutenue Doctorat de l’Université de Haute-Alsace par : Sarah ALAVI Spécialité : Chimie Organique Le jeudi 28 novembre 2013 Synthèse et évaluation biologique de dérivés de l’amino-benzosubérone, inhibiteurs puissants et sélectifs de l’Aminopeptidase N ou CD13. Ecole Doctorale : Jean Henri-Lambert 494 – Pôle Chimie, Physique et Matériaux Unité de recherche : Laboratoire de Chimie Organique et Bioorganique (COB) – EA 4566 Directeurs de Thèse : Pr Céline TARNUS (COB – UHA Mulhouse) Dr Philippe BISSERET (COB – UdS Strasbourg) Rapporteurs : Dr Jean-Bernard BEHR (ICMR – Univ. Champagne-Ardenne Reims) Dr Siden TOP (Lab.
    [Show full text]
  • Genome Sequencing and Functional Genes Comparison Between Sphingopyxis
    bioRxiv preprint doi: https://doi.org/10.1101/2021.03.29.437629; this version posted March 31, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 Genome sequencing and functional genes comparison between Sphingopyxis 2 USTB-05 and Sphingomonas morindae NBD5 3 4 Chao Liu1, Qianqian Xu1, Zhenzhen Zhao1, Shahbaz Ahmad1, Haiyang Zhang1, Yufan 5 Zhang1, Yu Pang1, Abudumukeyiti Aikemu1, Yang Liu1,* Hai Yan1,* 6 7 1 School of Chemistry and Biological Engineering, University of Science and 8 Technology Beijing, Beijing 100083, China 9 *Corresponding author: Prof Hai Yan; mail: [email protected]. Yang Liu; mail: 10 [email protected]. 11 12 Keywords: Sphingomonadaceae; Genome; Lutein; Hepatotoxin; Microcystins; 13 Biodegradation 14 15 16 17 18 19 20 21 22 23 24 25 1 bioRxiv preprint doi: https://doi.org/10.1101/2021.03.29.437629; this version posted March 31, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 26 ABSTRACT 27 Sphingomonadaceae has a large number of strains that can biodegrade hepatotoxins or 28 environmental pollutants. The latest research reported that certain strains can also 29 produce lutein. Based on the third-generation sequencing technology, we analyzed the 30 whole genome sequence and compared related functional genes of two strains of 31 Sphingomonadaceae isolated from different habitats. The genome of Sphingopyxis 32 USTB-05 was 4,679,489 bp and contained 4312 protein coding genes.
    [Show full text]
  • Tássio Brito De Oliveira Fungos Na Compostagem Da
    UNIVERSIDADE ESTADUAL PAULISTA “JÚLIO DE MESQUITA FILHO” unesp INSTITUTO DE BIOCIÊNCIAS – RIO CLARO PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIAS BIOLÓGICAS (ÁREA: MICROBIOLOGIA APLICADA) TÁSSIO BRITO DE OLIVEIRA FUNGOS NA COMPOSTAGEM DA TORTA DE FILTRO: DIVERSIDADE, GENÔMICA E POTENCIAL BIOTECNOLÓGICO Tese apresentada ao Instituto de Biociências, do Câmpus de Rio Claro, Universidade Estadual Paulista, como parte dos requisitos para obtenção do título de Doutor em Ciências Biológicas (Área: Microbiologia Aplicada). Rio Claro Dezembro – 2016 TÁSSIO BRITO DE OLIVEIRA FUNGOS NA COMPOSTAGEM DA TORTA DE FILTRO: DIVERSIDADE, GENÔMICA E POTENCIAL BIOTECNOLÓGICO Tese apresentada ao Instituto de Biociências, do Câmpus de Rio Claro, Universidade Estadual Paulista, como parte dos requisitos para obtenção do título de Doutor em Ciências Biológicas (Área: Microbiologia Aplicada). Orientador: Prof. Dr. André Rodrigues Co-orientadora: Profª. Drª. Eleni Gomes Rio Claro Dezembro – 2016 589.2 Oliveira, Tássio Brito de O48f Fungos na compostagem da torta de filtro : diversidade, genômica e potencial biotecnológico / Tássio Brito de Oliveira. - Rio Claro, 2016 127 f. : il., figs., gráfs., tabs., fots. Tese (doutorado) - Universidade Estadual Paulista, Instituto de Biociências de Rio Claro Orientador: André Rodrigues Coorientadora: Eleni Gomes 1. Fungos. 2. Pirosequenciamento. 3. Cana-de-açúcar. 4. Enzimas lignocelulolíticas. 5. Tratamento de resíduo. 6. Fungos termofílicos. I. Título. Ficha Catalográfica elaborada pela STATI - Biblioteca da UNESP Campus de Rio Claro/SP AGRADECIMENTOS Ao meu orientador, Dr. André Rodrigues, quem muito admiro pela competência profissional e genialidade, pela oportunidade, dedicação e incentivo à pesquisa durante sua orientação, que me abriu caminhos e diversas colaborações, permitindo meu crescimento profissional e pessoal. À minha co-orientadora, Drª.
    [Show full text]
  • Microcystins and Nodularin) by Novel Gram-Positive Bacteria
    OpenAIR@RGU The Open Access Institutional Repository at Robert Gordon University http://openair.rgu.ac.uk Citation Details Citation for the version of the work held in ‘OpenAIR@RGU’: WELGAMAGE DON, A. C. D., 2012. An investigation into the biodegradation of peptide cyanotoxins (microcystins and nodularin) by novel gram-positive bacteria. Available from OpenAIR@RGU. [online]. Available from: http://openair.rgu.ac.uk Copyright Items in ‘OpenAIR@RGU’, Robert Gordon University Open Access Institutional Repository, are protected by copyright and intellectual property law. If you believe that any material held in ‘OpenAIR@RGU’ infringes copyright, please contact [email protected] with details. The item will be removed from the repository while the claim is investigated. AN INVESTIGATION INTO THE BIODEGRADATION OF PEPTIDE CYANOTOXINS (MICROCYSTINS AND NODULARIN) BY NOVEL GRAM-POSITIVE BACTERIA By Welgamage Don Aakash Channa Dharshan A thesis submitted in partial fulfilment for the degree of Doctor of Philosophy Robert Gordon University February 2012 i TABLE OF CONTENTS Title page .............................................................................................. i Contents ............................................................................................... ii Declaration .......................................................................................... iii Dedication ........................................................................................... iv Acknowledgements ...............................................................................
    [Show full text]
  • Balsano Evelyn.Pdf
    Uptake, physiological responses and technical application of the aquatic fungus Mucor hiemalis EH5 for the removal of cyanobacterial toxins vorgelegt von MSc Evelyn Balsano geb. in Bozen, Italien von der Fakultät III – Prozesswissenschaften der Technischen Universität Berlin zur Erlangung des akademischen Grades Doktor der Naturwissenschaften - Dr. rer. nat. - genehmigte Dissertation Promotionsausschuss: Vorsitzender: Prof. Dr. Juri Rappsilber Gutachter: Prof. Dr. Stephan Pflugmacher Lima Gutachter: Prof. Dr. Enamul Hoque Gutachterin: Prof. Dr.–Ing. Vera Meyer Tag der wissenschaftlichen Aussprache: 05. Oktober 2017 Berlin 2017 Auch eine Reise von tausend Meilen beginnt mit dem ersten Schritt ACKNOWLEDGEMENTS This study was conducted at the Department of Ecological Impact Research and Ecotoxicology and was funded by the Elsa-Neumann-Scholarship (formally known as Nachwuchsförderungsgesetz-NaFöG) and in part by the Studienbeihilfen für postuniversitäre Ausbildungen, Autonome Provinz Bozen Südtirol. I especially thank my academic supervisor Prof. Dr. Stephan Pflugmacher Lima for the opportunity to work in his well-equipped department on such a fascinating and innovative topic. I am thankful for the guidance and advice during the years and his confidence in my work. I further very much appreciated the possibility to present the Green Liver Systems on the exhibition IFAT in Munich, the inauguration of Felidae in Barnim and other occasions at the TU Berlin, thereby offering me the opportunity of being also involved in other fruitful outcomes of our research group. I am deeply grateful to my co-supervisor Prof. Dr. Enamul Hoque for the constant support and interest in the progress of my thesis throughout all these years. I am especially thankful for always welcoming me at the Helmholtz Zentrum München as well as introducing me to his wonderful family.
    [Show full text]
  • Supplemental Materials and Methods
    Supplemental Materials and Methods Cell isolation and in vitro stimulation Human monocytes and neutrophils were isolated from fresh peripheral blood using RosetteSep™ Human Monocyte Enrichment Cocktail (STEMCELL Technology) and Polymorphophrep solution (Axis-Shield), respectively. Fresh cells (1 x 106) resuspended in complete RPMI 1640 medium without penicillin and streptomycin were plated into 96-well U- bottom plates, and then infected with live BPZE1 at a bacterium-to-cell ratio of 10:1. Cells and bacteria were centrifuged to facilitate contact at 2200 rpm for 2 min, followed by incubation for 2h at 37°C in a 5% CO2 atmosphere. Afterwards, polymyxin B (5 µg/ml) was added to kill extracellular bacteria. Then cells were extensively washed and further incubated for 6h or 12h. Cell activation was evaluated using flow cytometric analysis. Supernatants were collected from cell culture for the measurement of cytokines. Bacterial strains, growing conditions and lysates preparation Vaccine strain BPZE1 and a virulent clinical B. pertussis strain BP611-98 were obtained from the Public Health Agency of Sweden. As for the lysates preparation, both B. pertussis strains were grown on charcoal agar plates supplemented with 10% horse blood for 5 days at 37°C. Bacteria were then scraped gently from the plates, suspended in PBS, and then filtered through a 40 µm cell strainer. Bacteria suspensions were centrifuged at 3500 rpm for 15min and pellets were collected. An appropriate volume of lysis buffer (8M urea, 500mM bicarbonate, PH=8.5) was used to lyse the bacteria on ice for 1h, followed by centrifugation and collection of supernatants. Protein concentration was determined using Pierce BCA Protein Assay Kit according to the manual.
    [Show full text]