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Leidiane Andreia Acordi Menezes SOURDOUGH BIOTECHNOLOGY

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Leidiane Andreia Acordi Menezes SOURDOUGH BIOTECHNOLOGY Leidiane Andreia Acordi Menezes SOURDOUGH BIOTECHNOLOGY FOR IMPROVING BREAD NUTRITIONAL PROPERTIES Tese submetida ao Departamento de Ciência e Tecnologia de Alimentos da Universidade Federal de Santa Catarina para a obtenção do grau Doutor em Ciência dos Alimentos. Orientador: Prof. Dr. Juliano De Dea Lindner Florianópolis 2019 Ficha de identificação da obra elaborada pelo autor através do Programa de Geração Automática da Biblioteca Universitária da UFSC. A ficha de identificação é elaborada pelo próprio autor Maiores informações em: http://portalbu.ufsc.br/ficha Leidiane Andreia Acordi Menezes SOURDOUGH BIOTECHNOLOGY FOR IMPROVING BREAD NUTRITIONAL PROPERTIES Esta Tese foi julgada adequada para obtenção do Título de Doutora em Ciência dos Alimentos e aprovada em sua forma final pelo Programa ... Local, 15 de março de 2019. ________________________ Prof.(a). Dr (a). Ana Carolina de Oliveira Costa Coordenadora do Curso Banca Examinadora: ________________________ Prof. Dr. Juliano de Dea Linder Orientador (UFSC) ________________________ Prof.(a) Dr. (a) Michele Rigon Spier Membro (UFPR) ________________________ Prof.(a) Dr. (a) Monica Gatti Membro (UNIPR) ________________________ Prof.(a) Dr. (a) Edna Regina Amante Membro (UFSC) ________________________ Prof.(a) Dr. (a) Amanda Bagolin do Nascimento Membro (UFPR) À minha nona Santina Giusti Acordi. (in memorian) À minha mãe Maria Alice Acordi. À minha irmã Luciene Acordi e ao meu sobrinho e afilhado Felipe Luiz Acordi Nascimento. ACKNOWLEDGEMETS I sincerely thank to everyone that make this project possible. To Federal University of Santa Catarina (UFSC) and to the Post Graduation Program in Food Science (PPGCAL), for accept me as Doctorate student. To Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), for the scholarship in Brazil and in Italy. To Professor Juliano, for everything. For accepting to advise me and for trusting me. Thank you for orientation, friendship, and for being an admirable person. To Ministério da Agricultura Pecuária e Abastecimento (MAPA/São José – SC), Dr. Heitor Daguer, MSc. Luciano Molognoni and Leandro Ploêncio, for the collaboration with the chromatographic analysis. To Neoprospecta/Florianópolis – SC for the contribution with metagenomic analysis. To Professor Dr. Rubens T. D. Duarte, Department of Microbiology, Immunology and Parasitology – UFSC, for collaborating with the metagenomic analysis and bioinformatics. To Professors DR. Norma Machado da Silva and Dr. Carlos Peres Silva, to Dr. Roseane Panini and MSc. Cristina Rios, from the Laboratory of Biochemistry – UFSC, for the collaboration with electrophoresis. To Professor Dr. Mariana S. Larraz Ferreira and to MSc Thais de Oliveira Alves, from the Post Graduation Program in Food and Nutrition, from the University do Estado do Rio de Janeiro (UNIRIO), for collaborating with the proteomic analysis. To UNIPR – Università Degli Studi di Parma, for accept me as external student. To professors Erasmo Neviani and Monica Gatti, for receiving me and making possible the doctoral stage in Italy. To Maria Luisa Savo-Sardaro and UNIPR team for the support and for the working hours. To UTFPR/Medianeira professors, technicians and friends. To UFSC professors, technicians and friends, of the laboratories of Bioprocesses, Food Technology, Biotechnology and Molecular Biology. To Catharina Costa, Nátaly Neves and Fabio Costa for friendship and for the collaboration with the analysis. To my family, my mother Maria Alice Acordi, my sister Luciene Acordi, my nephew Felipe Luiz Acordi Nascimento and my Godmother Adenir Accordi, for always encouraging me to continue studying, and for being the light of my existence. To Juliane(a), Ana Beatriz, Mariana(s), Fernanda, Catiussa, Denise, Pâmela, Nívea, Daiany, Camila, Gabriela, Natália and Catharina for being my partners of life and soul. To my friends and athletes of the Desterro Atlantis Flag Football and Floripa Va’a. To my childhood friends, my friends from Cazona, and all my friends from Florianópolis, Santa Terezinha de Itaipu, Medianeira and Foz do Iguaçu. Every day I woke up sad, you raised me up. I'm very grateful to everyone. Non c’è bisogno di un motivo per tutto ciò che fai nella tua vita. Fallo perché vuoi farlo. Perché è divertente. Perché ti rende felice. Autore sconosciuto RESUMO A fermentação sourdough tem sido estudada por sua capacidade de melhorar as propriedades sensoriais, nutricionais e tecnológicas de pães, resultado do metabolismo da comunidade microbiana do sourdough, composta principalmente por bactérias ácido-láticas (BAL) e leveduras. O objetivo deste estudo foi caracterizar a evolução da comunidade microbiana de sourdoughs produzidos no Brasil durante as etapas de preparo (propagação) e investigar os efeitos da fermentação sourdough na concentração de FODMAPs (do inglês, Fermentable Oligosaccharides, Disaccharides, Monosaccharides and Polyols) em pães. FODMAPs são uma classe de carboidratos associados ao desencadeamento dos sintomas da Síndrome do Intestino Irritável (SII) e da Sensibilidade Não Celíaca ao Glúten (SNCG). Duas temperaturas foram testadas para propagação, 21 ºC ± 1ºC (SD1) e 30 ºC ± 1ºC (SD2). A composição microbiana dos sourdoughs foi investigada por métodos dependentes e independentes de cultivo. A concentração de FODMAPs foi avaliada por métodos enzimáticos e por Cromatografia Líquida acoplada à Espectrometria de Massas (LC/MS-MS). A comunidade microbiana da farinha de trigo foi composta exclusivamente por Proteobacteria (Escherichia hermannii, Kosakonia cowanii, Pantoea ananatis e algumas espécies de Pseudomonas). BAL foram detectados somente após a primeira etapa de propagação (BS1) e tornaram-se predominantes (Lactobacillus farciminis e Lactobacillus curvatus) após a quarta etapa (BS4). A partir desta etapa, a diferença entre as temperaturas afetou a dinâmica microbiana entre os dois tratamentos. Ao final, a comunidade microbiana do SD1 foi composta exclusivamente por BAL (L. farciminis, L. curvatus, Leuconostoc citreum e Pediococcus pentosaceus). Bacillus, Pseudomonas, Enterococcus e enterobacteria foram inibidos gradualmente durante a propagação e não foram detectados na etapa final. Por outro lado, a temperatura de 30 ºC ± 1ºC favoreceu a presença de bactérias atípicas (que não BAL) para o SD2. L. farciminis e Escherichia hermannii foram dominantes. Clostridium, Bacillus, Enterococcus foram inibidos, no entanto, Kosakonia cowanii, Pantoea ananatis e algumas espécies de Pseudomonas ainda estavam presentes ao final da propagação. Em relação aos FODMAPs, sacarose, frutose e glicose foram completamente consumidas a partir da primeira etapa de fermentação, enquanto os demais subgrupos dos FODMAPs tiveram suas concentrações reduzidas após a terceira e quarta etapas de fermentação, coincidindo com o predomínio da BAL na comunidade microbiana. Os pães fermentados por sourdough apresentaram maiores proporções de ácidos orgânicos e menor concentração de sacarose, frutose, glicose e frutanos comparados aos pães fermentados por Saccharomyces cerevisiae. A concentração de frutanos, o subgrupo mais importante de FODMAPs, foi de 40 a 62% menor nos pães sourdough. Portanto, a fermentação sourdough representa uma alternativa promissora para o desenvolvimento de pães que possam ser consumidos por indivíduos que possuam SSI e/ou SNCG. Palavras-chave: Sourdough. FODMAPs. Frutanos. Bactérias Ácido- láticas. Temperatura. Dinâmica Microbiana. RESUMO EXPANDIDO Introdução Alimentos derivados de cereais são componentes importantes da dieta, dentre os quais, destaca-se o pão, um dos principais alimentos consumidos diariamente em todas as partes do mundo. A fermentação de pães tem sido praticada ao longo de milênios, por meio do sourdough, uma mistura de farinha e água, fermentada por bactérias ácido-láticas (BAL) e leveduras. Inúmeros estudos têm confirmado a capacidade da fermentação sourdough para promover modificações benéficas nas propriedades nutricionais, tecnológicas e sensoriais de pães. A contribuição do sourdough na qualidade do pão é decorrente das conversões microbianas e enzimáticas ocorridas durante o preparo da massa e dependente do metabolismo da comunidade microbiana presente no processo de fermentação. Mais de 80 espécies de bactérias já foram identificadas em sourdoughs em diversas partes do mundo. No entanto, um estudo deste caráter ainda não havia sido realizado no Brasil. A atividade metabólica da comunidade microbiana afeta macro e micronutrientes dos pães, dentre as quais, a fermentação dos carboidratos é considerada a modificação mais expressiva. O termo FODMAPs (do inglês, Fermentable Oligosaccharides, Disaccharides, Monosaccharides and Polyols) é utilizado para descrever carboidratos fermentáveis de cadeia curta, um grupo heterogêneo de moléculas mal absorvidas durante o processo digestivo, com um amplo efeito sobre os processos gastrointestinais, incluindo-se o desencadeamento dos sintomas da Síndrome do Intestino Irritável (SII). Além disso, a ingestão de FODMAPs tem sido recentemente associada ao desencadeamento dos sintomas da Sensibilidade ao Glúten Não Celíaca (SGNC). As manifestações clínicas para ambas as condições são similares e incluem distensão e dor abdominal, diarreia, flatulência e mudança no padrão evacuatório, fadiga e depressão. Enquanto a SII acomete de 7 a 21% da população global, a SGNC afeta cerca de 0,6 a 6%, sendo frequente que um paciente apresente as duas condições simultaneamente. Apesar da conversão de hexoses e pentoses em ácidos orgânicos, inerente ao processo fermentativo,
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