DOCSLIB.ORG

(12) United States Patent (10) Patent No.: US 9,359,581 B2 Ness Et Al

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
(12) United States Patent (10) Patent No.: US 9,359,581 B2 Ness Et Al US009359581B2 (12) United States Patent (10) Patent No.: US 9,359,581 B2 Ness et al. (45) Date of Patent: *Jun. 7, 2016 (54) BIOTRANSFORMATION USING CI2N 5/8 (2006.01) GENETICALLY MODIFIED CANDIDA CI2N 9/04 (2006.01) (52) U.S. Cl. (71) Applicant: Synthezyme, LLC, Rensselaer, NY CPC .............. CIIC3/006 (2013.01); C12N 9/0006 (US) (2013.01); C12N 15/815 (2013.01): CI2P 7/6409 (2013.01): CI2P 7/649 (2013.01): CI2P (72) Inventors: Jon E. Ness, Redwood City, CA (US); 7/6427 (2013.01); C12P 7/6436 (2013.01); Jeremy Minshull, Palo Alto, CA (US) Y02E 50/13 (2013.01); Y02P 20/52 (2015.11) (58) Field of Classification Search (73) Assignee: Synthezyme LLC, Rensselaer, NY (US) None See application file for complete search history. (*) Notice: Subject to any disclaimer, the term of this patent is extended or adjusted under 35 (56) References Cited U.S.C. 154(b) by 0 days. U.S. PATENT DOCUMENTS This patent is Subject to a terminal dis claimer. 8, 158,391 B2 * 4/2012 Gross et al. ................... 435.134 (21) Appl. No.: 14/084.230 OTHER PUBLICATIONS Chica et al. Curr Opin Biotechnol. Aug. 2005:16(4):378-84.* (22) Filed: Nov. 19, 2013 Sen et al. Appl Biochem Biotechnol. Dec. 2007: 143(3):212-23.* (65) Prior Publication Data * cited by examiner US 2015/OO94483 A1 Apr. 2, 2015 Primary Examiner — Christian Fronda Related U.S. Application Data (74) Attorney, Agent, or Firm — Laurence P. Colton; Smith Tempel Blaha LLC (60) Division of application No. 12/775,306, filed on May 6, 2010, now Pat. No. 8,597,923, which is a (57) ABSTRACT SENNES applit s 2,672). A substantially pure Candida host cell is provided for the yo, s • L vs. 8 Y-s u a- als--- u • biotransformation of a substrate to a product wherein the host (60) Provisional application No. 61/176,064, filed on May cell is characterized by a first genetic modification class that 6, 2009. comprises one or more genetic modifications that collectively or individually disrupt at least one alcohol dehydrogenase (51) Int. Cl. gene in the Substantially pure Candida host cell. CI2P 7/64 (2006.01) CIIC3/00 (2006.01) 33 Claims, 28 Drawing Sheets U.S. Patent Jun. 7, 2016 Sheet 1 of 28 US 9,359,581 B2 Fatty Acid rt-NS Fatty Acid Fatty Acid CYP52AType P450 s COAOxidase O-HO-Fatty Acid (B-unsaturated fatty acyl CoA Fatty Alcohol Oxidase CYP52A Type P450 Hydratase OCH-Fatty Acid 3-hydroxy fatty-acyl CoA Dehydrogenase CYP52A Type P450 Dehydrogenase HOOC-Fatty Acid B-OXO fatty-acyl CoA Thiolase Fatty-acyl COA+ Acetyl CoA Figure U.S. Patent Jun. 7, 2016 Sheet 2 of 28 US 9,359,581 B2 Fatty Acid a-Oxidation s B-Oxidation s s is s 8. i. Fatty Acid Fatty Acid CYP52AType P450 X ApOX4 ApOx5 (strain DP1) y C-HO-Fatty Acid 0.8-unsaturated fatty acyl COA Fatty Alcohol Oxidase CYP52A Type P450 Hydratase y OCH-Fatty Acid 3-hydroxy fatty-acyl CoA Dehydrogenase CYP52A Type P450 Dehydrogenase s HOOC-Fatty Acid 8-OXO fatty-acyl CoA Thiolase Fatty-acyl CoA+ Acetyl CoA Figure2 U.S. Patent Jun. 7, 2016 Sheet 3 of 28 US 9,359,581 B2 albicans DH 1A MSLFRIFRGASLTTTTASFTATTGATT.KTLS GSTWLRKSYKRTYSSSWLSSFE albicans ADH 1E MERFFRIFEGGSLTTTTSFTTTGTTTLSGSTWLREEERTESSSWLSSPE tropicalis ADE 4 albicans ADH 24. albicans ADH 2B tropicalis ADH B4 trialis DH 1 tropicalis ADHB11 albicans DH 1A LFFFHFINNERYCHTTTTTNTRTIMSEIFETEAWWFDTNGGOLWYKDYPWPTFEERIE albicans DH 1E LFFFHOFINNERYCHTTTTTNTETIMSEQIPETEAWWFDTNGGOLWYEDYFWPTFEPTE tropicalis ADH 44 ELEYEDIPWRTREME albicans ADH 2A MSWFTTE WIFETIGGELEYEDIFFFERENE albicans DH 2B MSWFTTEAWIFETNGGKLEYEDIPWPKPKANE tropicalis ADH B4 LEEDIPWREPERIE ... tropicalis ADH 10 . KLEYEDWPWPWPEPE tropicalis ADHB11 FLYTDIPWPWPKFNE it fit fit fit, it albicans DH 1A LLIHESGHTILHWEGDWPLTELPLGGHEGGGGEEGWEIGDFGIEW albicans DE 1B LLINESSWCHTILHiWRGITVPLTELPLWGGHEGGGGEEGWEIGIF,GIEW tropicalis ADH 4 LLINKSGHTILHWEGIWFLTELFLGGHEGGGGEEGWEIGDFGIEW allian8 ADH 2. LLINESGHTILHWEGIWFLTELFLGGHEGFLGENEGWEGD, albicans ADH2B LLINESGCHTILHWRGITFLTELFLGGHEGGS WILGEMWKGWEGIGWEW ... tropicalis DH B4 LLINESGCHTILHWEGIWFLITELPLGGHEGGIGINEGWEGILGW tropicalis ADH.10 L LRESGCHSDLHTTEGDWFIFELFLGGHEGGGMGNREGTEGDLGIET tropicalis ADH Bll LLYHWESGWCHSDIHWWEGIWFPSELFWGGHEGGWWIGENWGWEWGDL.GIEM * f : . it it if it if f : * : * * * * * * : * * * : h if f it it if it it. if it; if it if f it: it albicans ADH 1A LNGSCMSCEFCOGEFNCGEDLSGTHDGSFETDAW, EIFGTILITAFIL albicans DH 1E LNGSCMSCEFCOGEFMCGEDLSGTHIGSFETDAWKIFAGTDLMWFIL tropicalis ADH 4 LNGSCMSCEFCOGEFMCGEDLSGTHIGSFETDAWRIPTILEWPIL albicans ADH2A LNGSCLNCEYCSGAEFNCEADLSGYTHIGSFQYATA DAN, RIPAGTDLANWAFIL albicans ADH 2B LNGSCLNCECSGAEFNCEDLSGTHIGSFYIT.I.W.RIPEGTILITAFIL ... tropicalis ADH B4 LNGSCMICEYCOOGEPNCPO, DLSGTHIGSFOOTDRIP, GTDLNWPIL tropicalis DH.10 LNGSCMICEFCOGAEFNCSRADMSG THDGTFOYATADAWKIPEGDMASIAFIL tropicalis ADH Bll LNGSCMNCEYCOGAEPMCPHDWSGSHDGTFYTD.W.EFPGSDLSIPIS * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * allians DH 1. CGTELETILGWISGGGGLGSLWORMGLRWIGGIEEGEFE albicans DH1B CGTWELETILGWISGGGGLGSLWYARMGLRWIDGGDEKGEFE tropicalis ADH 4 CGTELETILGISGGGLSLOWMGLRIGGIEEGFE albicans DH 2A C.G.T.YKALETELEGOWAISG. AGGLGSLWYAKMGYRLAIDGGEDKGEFWES albicans DH 2E CGITELETELEGISGIGGLGSLW.E.MGR LIDGGEDEGEFE Figure 3A U.S. Patent Jun. 7, 2016 Sheet 4 of 28 US 9,359,581 B2 C topicalis ADE E4 GWTWELETEDLFNISG, GLSLWEMGRWIDGGEGEFE C tropicalis ADE 10 CAGSTWELENADLLAGOT...ISGGGGLGSLGWE, MGRWLAIDGGIERGEFWE , tropicalis ADE B11 GWTWELET GLOFWISG, GLSLEMGLRWIGGERFES * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * C albicans ADH 14. LGENDFTEDEDIWEWEETIGGRHGINESSEEIDSWEWRFLGWWLWGLP. albicans ADH 1E LG EASWDFTEIRDIYESERTIGGPHGAINSYSERIDSWEEWRFLGRWLWGLFA topicalis ADE 44 LEIDFLEEEDIWSWEETIGGFHINESSEEISEWRFLGELLP albicans ADH2A LGETFIDFTEEEDWWE WEETINGGPHGWINSWSERIGOSTEYWRTLGEWWLWGLPA albicans ADH2E LGETFIDFTEEEDWWE WEETINGGFHGWINSWSERIGOSTE WRTLG&WLWGLFA tIOpicalis ADF E4 LGAEWFWDFL.EEEDIWGWEKATIGGRHGWNYSISEEINSWDWRTLGEWWLWGLFA , tropicalis DH 1. LGAEYYIDFLEEDIYSIRRATGGGFHGWINSWSEEINSWEWRTLGKWWLWSLFA tropicalis ADEE 11 LGAEWFWDFTEEANYSEIIRATIGGAHGSINESISEEINSWEEWRTLGTWLWGLPA * * * * * * * : ; ; t; fift fit; if it: k, t, if it, if t if f it? C albicans ADH 1A HETPWFIWWESIEIGSEGNREITEIDFFSRGLIECPIKIWGLSDLPEFELM C albicans ADH 1E HETPWFD WESIEIRGSGNREDTEIDFFSRGLIECPIEIWGLSDLPEFELE C tIOpialis DH GSEWTGWFEWWESIEIEGSYWGNREDTENDFFSRGLIECFIEIWGLSELFWFELM C albicans TH 2. GART3TPWFD WIFTIOIRGSYWGNRRDTAE ANDFFTRG, IRCFTRIWGLSELPEWYRLY allicans ADH2E GAEISTFWFD WIETIIEGSYNGNREDTAETIFFTRGLIECFIKIWGLSELFEWELE C , tropicalis ADF E4 GSEWSFWFDSWWESIQIEGSYWGNREDTESDFFSRGLIECFIEWGLSELFEWYELM C tIOpicalis ADE 10 GGELTPLFESWARSIOIRTTCWGNREDTTEIDFFWRGLIDCPIKWGLSEWPEIFILM tropicalis ADE B11 ELEFIFMSAEIIRGSYNNRRDTAEWDFFARGLWECFIEWGLSELFEIFELL f; ; ; ; ; ; ; ; f : h : h if it; it fifth if it? fift fit fift f ; ; ; ; ; ; , f : C. albicans ADH 1, EEGEILSRYWLITS-SEC ID MO; 72 C. albicans ADH 1B EEGRILGRYWLITSE SEC ID MO; 173 C. tropicalis ADH4 --------------- SE ID MO; 155 C. albicans ADH2. EEGRILGRYWLINDE SEO, ID MO; 174 C, albicans ADH 2B EEGKILGRLINK SE II MO; 175 C. tropicalis ADH E4 -------------- SE ID MO;154 C. tropicalis ADH 10 -------------- SE ID MO; 152 C. tropicalis ADH E 11 -------------- SE II MO; 151 Figure3B U.S. Patent Jun. 7, 2016 Sheet 5 of 28 US 9,359,581 B2 ReCOmbinase ReCOmbinase Site Site Targeting Recombinase Targeting Sequence enCOding gene Selective marker Sequence Restriction site to release targeting Restriction site to release targeting Construct from Sequences required for Construct from Sequences required for bacterial propagation bacterial propagation Figure 4 U.S. Patent Jun. 7, 2016 Sheet 6 of 28 US 9,359,581 B2 ReCOmbinase ReCOmbinase site Targeting COnstruct site Targeting Recombinase Targeting Sequence 1 enCOding gene Selective marker Z8 : TargetW: TargetNEB Sequence 1 Sequence 2 GenOmic DNA DNA region to be replaced HomologOUS recombination Integrant W ''''''''''''''''''''''^ ReCOmbinase GenOmic DNA enCOding gene Selective marker ReCOmbinase ReCOmbinase Site Site EXCised DNA W: N D Figure 5 U.S. Patent Jun. 7, 2016 Sheet 7 of 28 US 9,359,581 B2 ReCOmbinase ReCOmbinase Site Site ReCOmbinase encoding gene Selective marker Z8E0 N A GenOmic DNA Induction of reCOmbinase ReCOmbinase WXNE B : GenOmic DNA ReCOmbinase Recombinase site encoding gene Selective marker U.S. Patent Jun. 7, 2016 Sheet 8 of 28 US 9,359,581 B2 Insertion Sequences ReCOmbinase ReCOmbinase site Site Targeting Recombinase Targeting Sequence encoding gene Selective marker Sequence WIO E.8%:W.III E: Restriction site to release targeting Restriction site to release targeting construct from sequences required for Construct from sequences required for bacterial propagation bacterial propagation Figure 7 U.S. Patent Jun. 7, 2016 Sheet 9 of 28 US 9,359,581 B2 Insertion ReCOmbinase ReCOmbinase Sequences Targeting COnstruct Site Site Targeting Recombinase Targeting Sequence? encoding gene Selective marker eCuence 2 W| EN A : Target Target Sequence Sequence 2 Genomic DNA DNA region to be replaced Homologous recombination Insertion Sequence Integrant EWIE8Ex ReCOmbinase GenOmic DNA enCOding gene Selective marker ReCOmbinase ReCOmbinase Site site EXCISec DNA U.S. Patent Jun. 7, 2016 Sheet 10 of 28 US 9,359,581 B2 ReCOmbinase ReCOmbinase Site Site ReCOmbinase encoding gene Selective marker EZIEEEx:N Genomic DNA \""Sequences Induction OfreCOmbinase Insertion Sequences ReCOmbinase WIEX NE B GenOmic DNA Recombinase ReCombinase site enCOding gene Selective marker Figure 9 U.S. Patent Jun. 7, 2016 Sheet 11 of 28 US 9,359,581 B2 Original genomic sequence Target Sequence
Load more

Recommended publications
  • (12) Patent Application Publication (10) Pub. No.: US 2015/0094483 A1 Ness Et Al
    US 2015 0094.483A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0094483 A1 Ness et al. (43) Pub. Date: Apr. 2, 2015 (54) BIOTRANSFORMATION USING Publication Classification GENETICALLY MODIFIED CANDDA (51) Int. Cl. (71) Applicants: Jon E. Ness, Redwood City, CA (US); CI2P 7/64 (2006.01) Jeremy Minshull, Palo Alto, CA (US) CIIC3/00 (2006.01) CI2N 5/8 (2006.01) (72) Inventors: Jon E. Ness, Redwood City, CA (US); (52) U.S. Cl. Jeremy Minshull, Palo Alto, CA (US) CPC ............. CI2P 7/6409 (2013.01); CI2N 15/815 (2013.01): CI IC3/006 (2013.01) (21) Appl. No.: 14/084.230 USPC ....................... 554/219; 435/254.22:554/213 (22) Filed: Nov. 19, 2013 Related U.S. Application Data (57) ABSTRACT (60) Division of application No. 12,775,306, filed O May A substantially pure Candida host cell is provided for the 6, 2010, now Pat. No. 8,597,923, which is a continu biotransformation of a substrate to a product wherein the host ation-in-part of application No. 12/436,729, filed on cell is characterized by a first genetic modification class that May 6, 2009, now Pat. No. 8,158,391. comprises one or more genetic modifications that collectively (60) Provisional application No. 61/176,064, filed on May or individually disrupt at least one alcohol dehydrogenase 6, 2009. gene in the Substantially pure Candida host cell. Patent Application Publication Apr. 2, 2015 Sheet 1 of 28 US 201S/0094.483 A1 Fatty Acid rt-NS Fatty Acid Fatty Acid CYP52A Type P450 s COAOxidase (HO-Fatty Acid CB-unsaturated fatty acyl CoA Fatty Alcohol Oxidase CYP52A Type P450 Hydratase OCH-Fatty Acid 8-hydroxy fatty-acyl CoA Dehydrogenase CYP32A Type P450 Dehydrogenase HOOC-Fatty Acid BOXO fatty-acyl CoA Thiolase Fatty-acyl CoA+ Acetyl CoA Patent Application Publication Apr.
    [Show full text]
  • Diversity and Bioprospection of Fungal Community Present in Oligotrophic Soil of Continental Antarctica
    Extremophiles (2015) 19:585–596 DOI 10.1007/s00792-015-0741-6 ORIGINAL PAPER Diversity and bioprospection of fungal community present in oligotrophic soil of continental Antarctica Valéria M. Godinho · Vívian N. Gonçalves · Iara F. Santiago · Hebert M. Figueredo · Gislaine A. Vitoreli · Carlos E. G. R. Schaefer · Emerson C. Barbosa · Jaquelline G. Oliveira · Tânia M. A. Alves · Carlos L. Zani · Policarpo A. S. Junior · Silvane M. F. Murta · Alvaro J. Romanha · Erna Geessien Kroon · Charles L. Cantrell · David E. Wedge · Stephen O. Duke · Abbas Ali · Carlos A. Rosa · Luiz H. Rosa Received: 20 November 2014 / Accepted: 16 February 2015 / Published online: 26 March 2015 © Springer Japan 2015 Abstract We surveyed the diversity and capability of understanding eukaryotic survival in cold-arid oligotrophic producing bioactive compounds from a cultivable fungal soils. We hypothesize that detailed further investigations community isolated from oligotrophic soil of continen- may provide a greater understanding of the evolution of tal Antarctica. A total of 115 fungal isolates were obtained Antarctic fungi and their relationships with other organisms and identified in 11 taxa of Aspergillus, Debaryomyces, described in that region. Additionally, different wild pristine Cladosporium, Pseudogymnoascus, Penicillium and Hypo- bioactive fungal isolates found in continental Antarctic soil creales. The fungal community showed low diversity and may represent a unique source to discover prototype mol- richness, and high dominance indices. The extracts of ecules for use in drug and biopesticide discovery studies. Aspergillus sydowii, Penicillium allii-sativi, Penicillium brevicompactum, Penicillium chrysogenum and Penicil- Keywords Antarctica · Drug discovery · Ecology · lium rubens possess antiviral, antibacterial, antifungal, Fungi · Taxonomy antitumoral, herbicidal and antiprotozoal activities.
    [Show full text]
  • Diversidade E Bioprospecção De Fungos Presentes Em Solos Antárticos
    UNIVERSIDADE FEDERAL DE OURO PRETO NÚCLEO DE PESQUISAS EM CIÊNCIAS BIOLÓGICAS PROGRAMA DE PÓS-GRADUAÇÃO EM BIOTECNOLOGIA ELDON CARLOS QUERES GOMES DIVERSIDADE E BIOPROSPECÇÃO DE FUNGOS PRESENTES EM SOLOS ANTÁRTICOS Orientador: Prof. Dr. Luiz Henrique Rosa Coorientadora: Profa. Dra. Valéria Martins Godinho OURO PRETO MINAS GERAIS - BRASIL 2017 UNIVERSIDADE FEDERAL DE OURO PRETO NÚCLEO DE PESQUISAS EM CIÊNCIAS BIOLÓGICAS PROGRAMA DE PÓS-GRADUAÇÃO EM BIOTECNOLOGIA ELDON CARLOS QUERES GOMES DIVERSIDADE E BIOPROSPECÇÃO DE FUNGOS PRESENTES EM SOLOS ANTÁRTICOS Dissertação apresentada à Universidade Federal de Ouro Preto, como parte das exigências do Programa de Pós-Graduação em Biotecnologia, para obtenção do título de Mestre em Biotecnologia, área de concentração Biotecnologia Industrial. OURO PRETO MINAS GERAIS – BRASIL 2017 3 Q35d Queres, Eldon Carlos Gomes. Diversidade e bioprospecção de fungos presentes em solos Antárticos [manuscrito] / Eldon Carlos Gomes Queres. - 2017. 66f.: il.: color; grafs; tabs; mapas. Orientador: Prof. Dr. Luiz Henrique Rosa. Coorientadora: Profª. Drª. Valéria Martins Godinho. Dissertação (Mestrado) - Universidade Federal de Ouro Preto. Pró-Reitoria de Pesquisa e Pós-Graduação. Núcleo de Pesquisas em Ciências Biológicas. Programa de Pós-Graduação em Biotecnologia. Área de Concentração: Biotecnologia Industrial. 1. Biotecnologia. 2. Fungos . 3. Fungos do solo. 4. Solos . 5. Microbiologia. I. Rosa, Luiz Henrique . II. Godinho, Valéria Martins . III. Universidade Federal de Ouro Preto. IV. Titulo. CDU: 606 Catalogação: www.sisbin.ufop.br i ii "Porque Deus tanto amou o mundo que deu o seu Filho Unigênito, para que todo o que nele crer não pereça, mas tenha a vida eterna. ” João 3:16 "Vocês são o sal da terra. Mas se o sal perder o seu sabor, como restaurá-lo? Não servirá para nada, exceto para ser jogado fora e pisado pelos homens.
    [Show full text]
  • Fungos Presentes Em Sedimentos Marinhos E Lacustres Da Antártica: Taxonomia, Diversidade E Bioprospecção De Metabólitos Bioativos
    Universidade Federal de Minas Gerais Programa de Pós-Graduação em Microbiologia Fungos presentes em sedimentos marinhos e lacustres da Antártica: taxonomia, diversidade e bioprospecção de metabólitos bioativos Mayara Baptistucci Ogaki Belo Horizonte, MG 2019 Universidade Federal de Minas Gerais Programa de Pós-Graduação em Microbiologia Fungos presentes em sedimentos marinhos e lacustres da Antártica: taxonomia, diversidade e bioprospecção de metabólitos bioativos Tese apresentada ao programa de Pós-Graduação em Microbiologia do Instituto de Ciências Biológicas da Universidade Federal de Minas Gerais. Orientador: Dr. Luiz Henrique Rosa Departamento de Microbiologia/ICB, UFMG Belo Horizonte, MG 2019 Colaboradores: Dr. Carlos Augusto Rosa Departamento de Microbiologia/ICB, UFMG Dr. Carlos Leomar Zani Dra. Tânia Maria de Almeida Alves Centro de Pesquisas René Rachou, FIOCRUZ/MG Dra. Rosemary Vieira Dr. Arthur Ayres Universidade Federal Fluminense, RJ Dr. Juan Manoel Lírio Instituto Antartico Argentino, Buenos Aires, Argentina 043 Ogaki, Mayara Baptistucci. Fungos presentes em sedimentos marinhos e lacustres da Antártica: taxonomia, diversidade e bioprospecção de metabólitos bioativos [manuscrito] / Mayara Baptistucci Ogaki. – 2019. 212 f. : il. ; 29,5 cm. Orientador: Dr. Luiz Henrique Rosa. Tese (doutorado) – Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas. Programa de Pós-Graduação em Microbiologia. 1. Microbiologia. 2. Regiões Antárticas. 3. Fungos. 4. Bioprospecção. 5. Biodiversidade. I. Rosa, Luiz Henrique. II. Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas. III. Título. CDU: 579 Ficha Catalográfica elaborada por Fabiane C. M. Reis – CRB6/ 2680 Dedicatória Aos meus maiores apoios, “minha família e amigos”. Agradecimentos Agradeço a todo corpo docente e funcionários do programa de pós-graduação em Microbiologia e ao apoio financeiro do CNPq e CAPES pela concessão de minha bolsa de estudos e pelos subsídios que permitiram a concretização desta tese.
    [Show full text]
  • Dissertação De Mestrado Diversidade De Fungos Decompositores De
    Universidade Federal de Minas Gerais Instituto de Ciências Biológicas Departamento de Microbiologia Dissertação de Mestrado Diversidade de fungos decompositores de madeira presentes em ecossistemas brasileiros: bioprospecção de espécies úteis em processo de biodegradação de poluentes aromáticos Barbara Muniz Soares Belo Horizonte 2013 Barbara Muniz Soares Diversidade de fungos decompositores de madeira presentes em ecossistemas brasileiros: bioprospecção de espécies úteis em processo de biodegradação de poluentes aromáticos Dissertação de mestrado apresentada ao Programa de Pós-Graduação em Microbiologia do Departamento de Microbiologia do Instituto de Ciências Biológicas da Universidade Federal de Minas Gerais como requisito parcial para obtenção do título de Mestre em Microbiologia. Orientador: Luiz Henrique Rosa Coorientador: Carlos Augusto Rosa 2013 Departamento de Microbiologia Instituto de Ciências Biológicas Universidade Federal de Minas Gerais Agradecimentos À Deus pela oportunidade de uma vida repleta de possibilidades e de aprendizado; Ao Prof. Carlos A. Rosa pela acolhida e ensinamentos. Agradeço as oportunidades que me foram concedidas, toda orientação profissional e cuidado dispensados a mim durante todos esses anos. É um grande privilégio fazer parte desta equipe de trabalho que tanto admiro; Ao Prof. Luiz H. Rosa pela orientação e confiança. Agradeço pela oportunidade de trabalhar com projetos extraordinários, pela paciência de ensinar e pelo apoio constante. Sua ajuda e solidez foram imprescindíveis para o desenvolvimento
    [Show full text]
  • Diversity and Distribution of Hidden Cultivable Fungi Associated with Marine Animals of Antarctica Fungal Biology
    Fungal Biology 123 (2019) 507e516 Contents lists available at ScienceDirect Fungal Biology journal homepage: www.elsevier.com/locate/funbio Diversity and distribution of hidden cultivable fungi associated with marine animals of Antarctica Valeria Martins Godinho a, Maria Theresa Rafaela de Paula a, Debora Amorim Saraiva Silva a, Karla Paresque b, Aline Paternostro Martins c, * Pio Colepicolo c, Carlos Augusto Rosa a, Luiz Henrique Rosa a, a Departamento de Microbiologia, Instituto de Ci^encias Biologicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil b Laboratorio de Bentologia, Universidade Federal de Alagoas, Maceio, AL, Brazil c Departamento de Bioquímica, Instituto de Química, Universidade de Sao~ Paulo, Sao~ Paulo, Brazil article info abstract Article history: In the present study, we surveyed the distribution and diversity of fungal assemblages associated with 10 Received 22 November 2018 species of marine animals from Antarctica. The collections yielded 83 taxa from 27 distinct genera, which Received in revised form were identified using molecular biology methods. The most abundant taxa were Cladosporium sp. 1, 17 April 2019 Debaryomyces hansenii, Glaciozyma martinii, Metschnikowia australis, Pseudogymnoascus destructans, Accepted 1 May 2019 Thelebolus cf. globosus, Pseudogymnoascus pannorum, Tolypocladium tundrense, Metschnikowia australis, Available online 14 May 2019 and different Penicillium species. The diversity, richness, and dominance of fungal assemblages ranged Corresponding Editor: Nik Money among the host; however, in general, the fungal community, which was composed of endemic and cold- adapted cosmopolitan taxa distributed across the different sites of Antarctic Peninsula, displayed high Keywords: diversity, richness, and dominance indices. Our results contribute to knowledge about fungal diversity in Antarctic ocean the marine environment across the Antarctic Peninsula and their phylogenetic relationships with species Extremophiles that occur in other cold, temperate, and tropical regions of the World.
    [Show full text]
  • Utilização Do Vírus Da Febre Amarela No Desenvolvimento De Ferramentas Para O Uso Em Diagnóstico Diferencial E Descoberta De Novos Antivirais Contra Flavivírus
    1 Universidade Federal de Pernambuco Centro de Ciências Biológicas Programa de Pós-Graduação em Genética Amanda Gomes de Oliveira Carvalho Utilização do vírus da febre amarela no desenvolvimento de ferramentas para o uso em diagnóstico diferencial e descoberta de novos antivirais contra flavivírus Recife 2015 ii Amanda Gomes de Oliveira Carvalho Utilização do vírus da febre amarela no desenvolvimento de ferramentas para o uso em diagnóstico diferencial e descoberta de novos antivirais contra flavivírus Tese apresentada ao Programa de Pós-Graduação em Genética da Universidade Federal de Pernambuco como parte dos requisitos exigidos para obtenção do título de Doutor em Genética. Orientadora: Laura Helena Vega Gonzalez Gil, PhD. Coorientador: Giovani Rota Bertani, PhD. Recife 2015 Catalogação na fonte Elaine Barroso CRB 1728 Carvalho, Amanda Gomes de Oliveira Utilização do vírus da febre amarela no desenvolvimento de ferramentas para o uso em diagnóstico diferencial e descoberta de novos antivirais contra flavivírus/ Amanda Gomes de Oliveira Carvalho– Recife: O Autor, 2015. 103 folhas : il., fig., tab. Orientadora: Laura Helena Vega Gonzalez Gil Coorientador: Giovani Rota Bertani Tese (doutorado) – Universidade Federal de Pernambuco. Centro de Ciências Biológias. Genética, 2015. Inclui bibliografia e anexos 1. Vírus- genética 2. Flavivírus 3. Agentes antivirais I. Gonzalez Gil, Laura Helena Vega (orientadora) II. Bertani, Giovani Rota (coorientador) III. Título 576.5 CDD (22.ed.) UFPE/CCB-2015-106 iii Amanda Gomes de Oliveira Carvalho Utilização do vírus da febre amarela no desenvolvimento de ferramentas para o uso em diagnóstico diferencial e descoberta de novos antivirais contra flavivírus Aprovado em 02 / 03 / 2015 Banca Examinadora: ____________________________________________ Dra. Laura Helena Vega Gonzalez Gil Centro de Pesquisas Aggeu Magalhães ____________________________________________ Dr.
    [Show full text]
  • Diversity, Distribution, and Ecology of Fungi in the Seasonal Snow of Antarctica
    microorganisms Article Diversity, Distribution, and Ecology of Fungi in the Seasonal Snow of Antarctica Graciéle C.A. de Menezes 1, Soraya S. Amorim 1,Vívian N. Gonçalves 1, Valéria M. Godinho 1, Jefferson C. Simões 2, Carlos A. Rosa 1 and Luiz H. Rosa 1,* 1 Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, Brazil; [email protected] (G.C.A.d.M.); [email protected] (S.S.A.); [email protected] (V.N.G.); [email protected] (V.M.G.); [email protected] (C.A.R.) 2 Centro Polar e Climático, Universidade Federal do Rio Grande do Sul, Porto Alegre 91201-970, Brazil; jeff[email protected] * Correspondence: [email protected]; Tel.: +55-31-3409-2749 Received: 13 September 2019; Accepted: 8 October 2019; Published: 12 October 2019 Abstract: We characterized the fungal community found in the winter seasonal snow of the Antarctic Peninsula. From the samples of snow, 234 fungal isolates were obtained and could be assigned to 51 taxa of 26 genera. Eleven yeast species displayed the highest densities; among them, Phenoliferia glacialis showed a broad distribution and was detected at all sites that were sampled. Fungi known to be opportunistic in humans were subjected to antifungal minimal inhibition concentration. Debaryomyces hansenii, Rhodotorula mucilaginosa, Penicillium chrysogenum, Penicillium sp. 3, and Penicillium sp. 4 displayed resistance against the antifungals benomyl and fluconazole. Among them, R. mucilaginosa isolates were able to grow at 37 ◦C. Our results show that the winter seasonal snow of the Antarctic Peninsula contains a diverse fungal community dominated by cosmopolitan ubiquitous fungal species previously found in tropical, temperate, and polar ecosystems.
    [Show full text]
  • The Yeast Handbook Carlos Rosa, Gábor Péter (Eds.)
    The Yeast Handbook Carlos Rosa, Gábor Péter (Eds.) Biodiversity and Ecophysiology of Yeasts With 40 Figures and 43 Tables Dr. Gábor Péter Professor Carlos Rosa Budapest University of Economic Departmento de Microbiologia Sciences and Public Administration ICB, CP 486 Faculty of Food Sciences Universidade Federal de Minas Gerais National Collection of Agricultural and Belo Horizonte, MG Industrial Microorganisms 31270-901 Somloi út 14-16 Brazil H-1118 Budapest Hungary Cover pictures by courtesy of: Vincent Robert, Centraalbureau voor Schimmelcultures (CBS), The Netherlands Carlos Rosa, Universidade Federal de Minas Gerais, Brazil ISBN-10 3-540-26100-1 Springer Berlin Heidelberg New York ISBN-13 978-3-540-26100-1 Springer Berlin Heidelberg New York Library of Congress Control Number: 2005930628 This work is subject to copyright. All rights are reserved, whether the whole or part of the mate- rial is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recita- tion, broadcasting, reproduction on microfilm or in any other way, and storage in data banks. Duplication of this publication or parts thereof is permitted only under the provisions of the German Copyright Law of September 9, 1965, in its current version, and permission for use must always be obtained from Springer. Violations are liable to prosecution under the German Copyright Law. Springer-Verlag is a part of Springer Science+Business Media springeronline.com © Springer-Verlag Berlin Heidelberg 2006 Printed in Germany The use of general descriptive names, registered names, trademarks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use.
    [Show full text]
  • TESIS390270 VAL.Pdf
    TESIS DOCTORAL María Isabel Pozo Romero 2012 YEASTS in FLORAL NECTAR: community ecology and interactions with insect pollinators and host plants TESIS DOCTORAL Yeasts in floral nectar: community ecology and interactions with insect pollinators and host plants Autora: María Isabel Pozo Romero Directores: Carlos M. Herrera y Conchita Alonso Estación Biológica de Doñana (CSIC). Departamento de Ecología Evolutiva. Sevilla, 2012 i TESIS DOCTORAL Yeasts in floral nectar: community ecology and interactions with insect pollinators and host plants Levaduras en el néctar floral: ecología de comunidades e interacciones con insectos polinizadores y plantas hospedadoras Memoria presentada por María Isabel Pozo Romero para optar al grado de Doctor por la Universidad de Sevilla Fdo. María Isabel Pozo Romero Vº Bº de los directores, Dr. Carlos M. Herrera Maliani Dr. María Concepción Alonso Menéndez Estación Biológica de Doñana, Estación Biológica de Doñana, Consejo Superior de Investigaciones Consejo Superior de Investigaciones Científicas Científicas Tutora de los cursos de doctorado, Dr. Montserrat Arista Palmero Universidad de Sevilla iii A mi familia. A Alfredo. v Agradecimientos Cuando empecé esta tesis, en 2007, ya erais muchos los “culpables” a los que agradecer mi incursión (o intromisión, una “ambientóloga”… ☺) en este mundo. Ahora, mientras escribo estas líneas, sólo puedo dejar constancia que el número de culpables ha crecido considerablemente, y que todos y cada uno de los citados -y los que me dejo en el camino, seguro- debéis atribuiros una parte considerable de este trabajo y, por qué no decirlo, de esta parte de mi vida. Empecemos. En primer lugar, quiero darle las gracias a mi director, Carlos.
    [Show full text]
  • Nectar-Living Yeasts of a Tropical Host Plant Community: Diversity and Effects on Community-Wide Floral Nectar Traits
    Nectar-living yeasts of a tropical host plant community: diversity and effects on community-wide floral nectar traits Azucena Canto1, Carlos M. Herrera2,* and Rosalina Rodriguez1,* 1 Centro de Investigacion Cientifica de Yucatan, A.C., Merida, Yucatan, Mexico 2 Estación Biológica de Doñana, CSIC, Sevilla, Spain * These authors contributed equally to this work. ABSTRACT We characterize the diversity of nectar-living yeasts of a tropical host plant community at different hierarchical sampling levels, measure the associations between yeasts and nectariferous plants, and measure the effect of yeasts on nectar traits. Using a series of hierarchically nested sampling units, we extracted nectar from an assemblage of host plants that were representative of the diversity of life forms, flower shapes, and pollinator types in the tropical area of Yucatan, Mexico. Yeasts were isolated from single nectar samples; their DNA was identified, the yeast cell density was estimated, and the sugar composition and concentration of nectar were quantified using HPLC. In contrast to previous studies from temperate regions, the diversity of nectar-living yeasts in the plant community was characterized by a relatively high number of equally common species with low dominance. Analyses predict highly diverse nectar yeast communities in a relatively narrow range of tropical vegetation, suggesting that the diversity of yeasts will increase as the number of sampling units increases at the level of the species, genera, and botanical families of the hosts. Significant associations between specific yeast species and host plants were also detected; the interaction between yeasts and host plants impacted the effect of yeast cell density on nectar sugars.
    [Show full text]
  • Kluyveromyces Siamensis Sp. Nov., an Ascomycetous
    RESEARCH ARTICLE Kluyveromyces siamensis sp. nov., an ascomycetous yeast isolated from water in a mangrove forest in Ranong Province,Thailand Somjit Am-In, Wichien Yongmanitchai & Savitree Limtong Department of Microbiology, Faculty of Science, Kasetsart University, Bangkok, Thailand Correspondence: Savitree Limtong, Abstract Department of Microbiology, Faculty of Science, Kasetsart University, Bangkok Seven strains of a novel Kluyveromyces species were isolated from seven water 10900, Thailand. Tel.: 166 2 562 5444, samples collected from a mangrove forest. Analysis of the D1/D2 domain of the ext. 4017; fax: 166 2 579 2081; e-mail: large subunit (LSU) rRNA gene sequences revealed that the sequences of five [email protected] strains (RS8T, RS20, RS54, RV42 and RV89) were identical and differed from the other two strains (RS65 and RV153) by only one nucleotide substitution in Received 29 January 2008; revised 28 March 544 nucleotides (nt). The closest species in terms of pairwise sequences similarity 2008; accepted 26 April 2008. was Kluyveromyces aestuarii, but the level of nucleotide substitution (six to seven First published online 6 June 2008. nucleotide substitutions in 544 nt) was sufficient to justify the description of a separate species. The phylogenetic tree based on the sequence of the D1/D2 DOI:10.1111/j.1567-1364.2008.00396.x domain of the LSU rRNA gene rather suggested that the new species is a sister Editor: Cletus Kurtzman species of K. aestuarii and forms a clade with the other six recognized species of Kluyveromyces. Sequence analysis of the internal transcribed spacer (ITS1– Keywords 5.8S rRNA gene–ITS2) region supported their distinct status as a species.
    [Show full text]