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Avaliação Do Potencial Nutricional, Antioxidante E Antibacteriano De Cogumelos Comestíveis

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Avaliação Do Potencial Nutricional, Antioxidante E Antibacteriano De Cogumelos Comestíveis UNIVERSIDADE FEDERAL DO PARANÁ FABIANE BACH AVALIAÇÃO DO POTENCIAL NUTRICIONAL, ANTIOXIDANTE E ANTIBACTERIANO DE COGUMELOS COMESTÍVEIS CURITIBA 2017 FABIANE BACH AVALIAÇÃO DO POTENCIAL NUTRICIONAL, ANTIOXIDANTE E ANTIBACTERIANO DE COGUMELOS COMESTÍVEIS Tese de Doutorado apresentada como requisito parcial à obtenção do grau de Doutora em Engenharia de Alimentos ao Programa de Pós- Graduação em Engenharia de Alimentos, Departamento de Engenharia Química, Setor de Tecnologia, Universidade Federal do Paraná. Orientador: Prof. Dr. Charles Windson Isidoro Haminiuk Co-orientadora: Drª. Cristiane Vieira Helm CURITIBA 2017 B118a Bach, Fabiane Avaliação do potencial nutricional, antioxidante e antibacteriano de cogumelos comestíveis / Fabiane Bach. – Curitiba, 2017. 135 f. : il. color. ; 30 cm. Tese - Universidade Federal do Paraná, Setor de Tecnologia, Programa de Pós- Graduação em Engenharia de Alimentos, 2017. Orientador: Charles Windson Isidoro Haminiuk. Coorientadora: Cristiane Vieira Helm. 1. Cromatografia líquida de ultra-eficiência. 2. Compostos fenólicos totais. 3. Perfil de aminoácidos. I. Universidade Federal do Paraná. II. Haminiuk, Charles Windson Isidoro. III. Helm, Cristiane Vieira. IV. Título. CDD: 613.2 Dedico este trabalho àqueles que tudo fizeram Para que eu me tornasse quem sou, E para que chegasse onde estou: Flavio, Tere, Felipe e Rudi! Agradecimentos Agradeço a Deus, por estar sempre presente e me permitir vida e saúde para a realização desse trabalho. Ao meu pai Flavio, meu herói, que todos os dias dá o exemplo de como ser o melhor ser humano que existe nesse mundo! À minha mãe, Teresinha, uma guerreira, meu exemplo de força e superação! Ao meu irmão, Felipe, por me orgulhar de suas conquistas e ser tão dedicado em tudo o que faz! Ao meu noivo, Rudimar, por todo o apoio, amor, amizade, conselhos, compreensão.... enfim, por ser meu porto seguro! Ao Prof. Dr. Charles W. I. Haminiuk, pela orientação e confiança em mim depositada e também pela compreensão nos momentos mais difíceis. À Drª. Cristiane V. Helm pela co-orientação, apoio, compreensão e confiança no meu trabalho. Ao Dr. Acácio A. F. Zielinski e à prof. Drª. Giselle M. Maciel, pelos conhecimentos repassados. Aos doutores membros da banca, por aceitarem participar da avaliação do trabalho. Aos professores do Programa de Pós Graduação em Engenharia de Alimentos da UFPR. À CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) pela bolsa de estudo concedida. À EMBRAPA (Empresa Brasileira de Pesquisa Agropecuária), UFPR (Universidade Federal do Paraná) e UTFPR (Universidade Tecnológica Federal do Paraná) pela infra- estrutura disponibilizada para a realização do trabalho. Aos colaboradores da Embrapa: Daiane, Dayanne, Edson, Fabrício, Janayna, Marianne e Tiélidy por todo auxílio e esclarecimento. Às técnicas de laboratório Andréa, Patrícia e Sheila, pela amizade e colaboração na realização desse trabalho. Às minhas queridas companheiras de laboratório: Aline, Bruna, Dayanne e Tamires, pela amizade, conversas e gargalhadas que facilitaram o trabalho. Aos meus amigos engenheiros Alessandra, Ana Paula, Elaine, Fernanda, Manoela, Marcelo e Thaísa, por estarem sempre dispostos a ajudar e compartilhar os momentos bons e ruins! Às minhas amigas de longa data: Camila, Crissie, Laís e Luana por permanecerem ao meu lado! Aos demais colegas do programa de pós-graduação em Engenharia de Alimentos, pelos momentos de descontração e trocas de experiências. Aos produtores de cogumelos comestíveis, Vânia (Campina Grande do Sul/PR), Shigueaki (Cornélio Procópio/PR) e Youko (Piedade/SP), por fornecerem algumas amostras que foram analisadas nesse trabalho. A todos aqueles cujos nomes não foram citados, mas que de alguma forma contribuíram para que esse trabalho fosse realizado. Muito obrigada! “É muito melhor lançar-se em busca de conquistas grandiosas, mesmo expondo-se ao fracasso, do que alinhar-se com os pobres de espírito, que nem gozam muito, nem sofrem muito, porque vivem numa penumbra cinzenta, onde não conhecem nem vitória, nem derrota.” Theodore Roosevelt RESUMO GERAL O consumo de cogumelos pela população mundial ocorre há milhares de anos, entretanto, atualmente, ainda não representa uma parcela significativa na dieta humana. Muitas espécies de cogumelos comestíveis são desconhecidas ou apresentam lacunas nas informações referentes ao seu potencial nutricional e nutracêutico. Os cogumelos são nutricionalmente equilibrados e possuem compostos biologicamente ativos que podem apresentar efeitos benéficos à saúde, como potencial antioxidante, antibacteriano, anti-inflamatório, antimutagênico, anticâncer, neuroprotetivo, hepatoprotetivo entre outros. As β-glucanas, polissacarídeos que constituem a fração solúvel das fibras alimentares dos cogumelos, são conhecidas por apresentarem atividade biológica. Além delas, os compostos provenientes do metabolismo secundário desses fungos, como os compostos fenólicos, também apresentam bioatividade (Capítulo 1). Devido a existência de lacunas no estudo do potencial nutricional e presença de compostos bioativos, em algumas espécies de cogumelos comestíveis, esta pesquisa teve por objetivo avaliar a composição físico-química e nutricional de sete espécies de cogumelos comestíveis comercializados no Brasil, otimizar a extração dos compostos fenólicos totais e estimar as atividades antioxidante e antibacteriana dos extratos fenólicos. Os cogumelos analisados foram: Agaricus bisporus (Champignon e Portobelo), A. brasiliensis, Flammulina velutipes, Lentinula edodes, Pleurotus djamor, P. eryngii, P. ostreatus (“Ostra” branca e “Ostra” preta). No Capítulo 2 as amostras foram analisadas quanto a sua composição centesimal, mineral, aminoacídica e fração das fibras solúveis (α e β-glucanas). Os resultados obtidos foram comparados com os padrões de ingestão dietética recomendada (RDA) e tratados utilizando a Análise de Componente Principal. Todos os cogumelos avaliados podem ser considerados como fonte potencial e alternativa de fibra alimentar (24,4 a 46,62%) e proteína (16,47 a 36,96%), contendo todos os aminoácidos essenciais à dieta humana, além de apresentar conteúdo de β-glucanas variando 1,58 a 16,91 mg/g de matéria seca (MS) e baixos teores de lipídeos (1,40 a 2,08%) e sódio (<66 mg/100 g MS). No Capítulo 3, realizou-se a otimização e modelagem da extração dos compostos fenólicos totais (CFT) de cada cogumelo. Os CFT variaram de 4,91 a 13,16 mg ácido gálico equivalente (GAE)/g MS. A partir do extrato fenólico otimizado, foram determinados os flavonoides totais (0,24 a 2,05 mg catequina equivalente (CE)/g MS), a atividade antioxidante pelos ensaios de ABTS (28,85 a 128,60 µmol trolox equivalente (TE)/g), DPPH (8,67 a 50,64 µmol TE/g) e FRAP (8,09 a 48,26 µmol TE/g), a determinação de compostos fenólicos por CLUE e a avaliação da atividade antibacteriana. Foram identificados 17 compostos nos extratos: os ácidos gálico, p-hidroxibenzoico, protocatecuico, gentísico, siríngico, vanílico, p-cumárico, ferúlico, clorogênico, cafeico, e trans-ciâmico; os flavonoides quercetina e campferol; e também os compostos catecol, vanilina e ácidos fumárico e benzoico. A atividade antimicrobiana foi testada usando as bactérias Staphilococcus aureus, Bacillus cereus, Escherichia coli e Salmonella enteretidis, pelo método de microdiluição. Todos os extratos fenólicos de cogumelos foram efetivos na inibição do crescimento das bactérias Gram-positivas (MIC ≤ 200 mg/mL), enquanto que para as bactérias Gram-negativas 66,7% dos extratos apresentaram atividade nas concentrações testadas. No Capítulo 4, realizou-se um estudo complementar para verificar a influência exercida pelo meio de cultivo [Quercus acutíssima (QA) e substrato axênico (SA)] sobre a composição físico-química do L. edodes. Os cogumelos produzidos em SA apresentaram aproximadamente 26% a mais de lipídeos se comparado ao cogumelo produzido em QA, 19% a mais de proteína, 5,3% a mais de carboidratos totais e 3,3% a mais de fibras solúveis. Em contrapartida, os L. edodes produzidos em QA apresentaram maior conteúdo de fibra alimentar (17% a mais) e β-glucanas (24% a mais). Palavras-chave: bioatividade, cromatografia líquida de ultra-eficiência, compostos fenólicos totais, perfil de aminoácidos, β-glucanas. ABSTRACT The mushrooms’s consumption by the world population occurs for thousands of years, however, currently, it does not yet represent a significant part of the human diet. Many species of edible mushrooms are unknown or lack information on their nutritional and nutraceutical potential. The mushrooms are nutritionally balanced and have biologically active compounds which can have beneficial effects on health as antioxidant potential, antibacterial, anti- inflammatory, antimutagenic, anticancer, neuroprotective, hepatoprotective among other potentials. The β-glucans, are polysaccharides of the soluble fiber fraction of mushrooms and They are known to have biological activity. Besides them, compounds from the secondary metabolism of these fungi, such as phenolic compounds, also present bioactivity (Chapter 1). Due to the existence of gaps in the study of the nutritional potential and the presence of bioactive compounds in some edible mushroom species, this study aimed to evaluate the physicochemical and nutritional composition of seven species of edible mushrooms marketed in Brazil, to optimize the extraction of total phenolic compounds and to estimate the antioxidant and antibacterial activities of phenolic extracts. The mushrooms analyzed were: Agaricus bisporus (Champignon and
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