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Biodiversidade De Leveduras Derivadas De Ecossistemas Antárticos Marinhos E Terrestres E Prospecção De Lipases

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Biodiversidade De Leveduras Derivadas De Ecossistemas Antárticos Marinhos E Terrestres E Prospecção De Lipases ALYSSON WAGNER FERNANDES DUARTE Biodiversidade de leveduras derivadas de ecossistemas Antárticos marinhos e terrestres e prospecção de lipases Tese apresentada ao Programa de Pós‐Graduação Interunidades em Biotecnologia USP/Instituto Butantan/IPT, para obtenção do Título de Doutor em Biotecnologia. São Paulo 2015 ALYSSON WAGNER FERNANDES DUARTE Biodiversidade de leveduras derivadas de ecossistemas Antárticos marinhos e terrestres e prospecção de lipases Tese apresentada ao Programa de Pós‐Graduação Interunidades em Biotecnologia USP/ Instituto Butantan/ IPT, para obtenção do Título de Doutor em Biotecnologia. Área de Concentração: Microbiologia Orientadora: Profa Dra. Lara Durães Sette Versão corrigida. A versão original eletrônica encontra‐se disponível tanto na Biblioteca do ICB quanto na Biblioteca Digital de Teses e Dissertações da USP (BDTD). São Paulo 2015 DADOS DE CATALOGAÇÃO NA PUBLICAÇÃO (CIP) Serviço de Biblioteca e Informação Biomédica do Instituto de Ciências Biomédicas da Universidade de São Paulo reprodução não autorizada pelo autor Duarte, Alysson Wagner Fernandes. Biodiversidade de leveduras de ecossistemas antárticos marinhos e terrestres e prospecção de lipases / Alysson Wagner Fernandes Duarte. -- São Paulo, 2014. Orientador: Profa. Dra. Lara Durães Sette. Tese (Doutorado) – Universidade de São Paulo. Instituto de Ciências Biomédicas. Programa de Pós-Graduação Interunidades em Biotecnologia USP/IPT/Instituto Butantan. Área de concentração: Biotecnologia. Linha de pesquisa: Microbiologia de ambiente extremo. Versão do título para o inglês: Biodiversity of marine and terrestrial yeasts from Antartic ecossystems and prospection of lipeases. 1. Levedura 2. Antártica 3. Lipase 4. Diversidade 5. Planejamento experimental 6. Sistema de duas fases aquosas I. Sette, Profa. Dra. Lara Durães II. Universidade de São Paulo. Instituto de Ciências Biomédicas. Programa de Pós-Graduação Interunidades em Biotecnologia USP/IPT/Instituto Butantan III. Título. ICB/SBIB0181/2014 UNIVERSIDADE DE SÃO PAULO Programa de Pós-Graduação Interunidades em Biotecnologia Universidade de São Paulo, Instituto Butantan, Instituto de Pesquisas Tecnológicas _____________________________________________________________________________________________________________ Candidato(a): Alysson Wagner Fernandes Duarte. Título da Tese: Biodiversidade de leveduras de ecossistemas antárticos marinhos e terrestres e prospecção de lipases. Orientador(a): Profa. Dra. Lara Durães Sette. A Comissão Julgadora dos trabalhos de Defesa da Tese de Doutorado, em sessão pública realizada a ................./................./................., considerou ( ) Aprovado(a) ( ) Reprovado(a) Examinador(a): Assinatura: .............................................................................................. Nome: ..................................................................................................... Instituição: .............................................................................................. Examinador(a): Assinatura: .............................................................................................. Nome: ..................................................................................................... Instituição: ............................................................................................... Examinador(a): Assinatura: .............................................................................................. Nome: ...................................................................................................... Instituição: ............................................................................................... Examinador(a): Assinatura: .............................................................................................. Nome: ...................................................................................................... Instituição: ............................................................................................... Presidente: Assinatura: .............................................................................................. Nome: ..................................................................................................... Instituição: .............................................................................................. Aos meus pais Jorge Fernandes Duarte e Maria de Lourdes Fernandes Duarte, pela força e fé com que levam a vida, acreditando sempre que um mundo melhor depende de nossas atitudes rotineiras!! Pelos exemplos de determinação, força, superação, carinho, amor! Por tudo que sou!!! A minha esposa Rafaela Tenório, por todos os momentos!! Por abdicar de parte de seus sonhos e embarcar comigo neste caminho!! Por todos os momentos de força, incentivo, espera e compreensão nessa caminhada! Te AMO!!! AGRADECIMENTOS A finalização de uma caminhada é feita ao lado de muitas pessoas importantes, sem as quais não seria possível a conclusão desse trabalho. Obrigado a todos que estiveram acompanhando esse trajeto. Aqui meus sinceros agradecimentos: À minha Orientadora Profa. Dra. Lara Durães Sette pelos ensinamentos, confiança, liberdade na discussão do trabalho e pela amizade! Obrigado por ter me possibilitado ir a este ambiente tão especial que é o Continente Antártico! A Divisão de Recursos Microbianos (DRM – CPQBA – UNICAMP) em nome das Profas. Dra. Valéria Maia de Oliveira, Dra. Fabiana Faninatti e Dra. Derlene Angelis por toda a infraestrutura e pela convivência durante o desenvolvimento deste trabalho; Às técnicas da Divisão de Recursos Microbianos, Éricka Letícia, Milena Binatti, Samantha Fonseca e Viviane Piccin pela pronta disposição em auxiliar os alunos da DRM, pela convivênvia e amizade. Vivi, obrigado por todos os momentos de auxílio e reflexão a respeito da identificação taxonômica das leveduras; Ao Centro Pluridicisplinar de Pesquisas Químicas, Biológicas e Agrárias (CPQBA – UNICAMP), em nome de toda a equipe técnica e administrativa, especialmente aos funcionários Célio, Celso, Laura, Lídia, Luíz, Paulo, Rita, Sandra e Valdir por todo o auxílio nos trâmites burocráticos, como a compra de reagentes ou no envio de malotes, ou trâmites rotineiros, como a utilização do restaurante universitário; Ao Prof. Dr. Adalberto Pessoa (Faculdade de Ciências Farmacêuticas – USP) pela parceria na abordagem dos Sistemas de Extração de Duas Fases Aquosas da lipase de L. scottii L117 e pela amizade; Ao Prof. Dr. Fernando C. Pagnocca e a Ifeloju Dayo-Owoyemi (UNESP – Rio Claro) pelo auxílio e discussão na avaliação taxonômica das leveduras; À Profa. Eleni Gomes, a Ana Ferrarezi e Rafaela Brito (UNESP – São José do Rio Preto) pelas discussões no planejamento de purificação da lipase de L. scottii L117; À equipe da Profa. Dra. Franciane Pellizzari (Universidade Estadual do Paraná) pela coleta e identificação taxonômica das macroalgas do ambiente Antártico; À equipe das Profas Dra. Maria Virginia Petri (Universidade do Vale do Rio dos Sinos) e Profa. Dra. Rosalinda Carmela Monton (Instituto Oceanográfico - USP), especialmente ao Caio Cipro pela coleta e identificação dos liquens do ambiente Antártico; À banca de qualificação pelas sugestões e conselhos em nome da Profa. Dra. Luiziana F. Silva (ICB – USP), Profa. Dra. Suzan P. Vasconcellos (UNIFESP) e Prof Dr. Wellington Luiz de Araújo (ICB – USP); À banca de defesa de tese em nome dos Professores Dr. Fernando C. Pagnocca (UNESP – Rio Claro), Dra. Eleonora Carmona (UNESP – Rio Claro), Dr. Itamar S. Mello (EMBRAPA - Jaguariúna) e Dr. Wellington L. de Araújo (ICB - USP) pelas sugestões e colaborações na discussão do trabalho; À Dra. Rafaella Bonugli (Universidade Federal da Integração Latino-Americana) pelo auxílio nas análises de Delineamento Experimental; Aos amigos André M. Lopes e João V. D. Molino (Laboratório de Biotecnologia Farmacêutica – USP) pelo auxilio no planejamento e discussão dos Sistemas de Extração de Duas Fases Aquosas da lipase de L. scottii L117; Aos amigos Prof. Dr. Michel Rodrigo Zambrani Passarini (Universidade Federal da Integração Latino- Americana) e Dr. Tiago Palladino (DRM – CPQBA) pelas discussões a respeito das ferramentas em estudos de diversidade; Aos amigos do Laboratório Divisão de Recursos Microbianos (DRM / CPQBA), por todos os momentos do café, pela descontração, convivência e pela amizade: Bruna, Cláudia, Daiane Daniela, Elmer, Fernando, Gileno, Gisele, Júlia, Leandro, Leonardo, Lucélia, Marcela, Mariana Nery, Maria Rapha, Michel, Milene, Meline, Natália, Nataly, Patrícia, Renan, Sanderson, Tiago Palladino, Tiago Rodrigues. Aos amigos que fiz na viagem ao continente gelado, especialmente aos Prof. Dr. Luiz H. Rosa (UFMG), Prof. Dr. Itamar S. Mello e João Luiz (Embrapa - Jaguariúna); Diego Catillo e Amanda Bendia (Institito Oceanográfico – USP); À família de D. Natalina Oliveira e Roberto Oliveira, nossa família paulista, por toda a acolhida quando chegamos em Valinhos. Por toda a confiança genuína em acolher pessoas “desconhecidas” e que foi um diferencial quando aqui chegamos!! Por todos os momentos, pela pronta disposição em auxiliar sempre, pela referência do que é verdadeiramente ser uma Família Cristã!!! Por todos os momentos com aquele café quentinho e uma conversa muito agradável; Ao Programa Brasileiro de Pesquisa na Antártica, em nome da Marinha do Brasil, Força Aérea Brasileira e Ministério de Ciência e Tecnologia pelo suporte e logística durante a viagem a Antártica; A toda equipe embarcada do navio Polar Almirante Maximiano H41 da Marinha do Brasil
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