Development of a defined synthetic medium to mimic sugarcane molasses for ethanol fermentation.

Detalhes bibliográficos
Autor(a) principal: Cunha, Gabriel Caetano de Gois e
Data de Publicação: 2022
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Biblioteca Digital de Teses e Dissertações da USP
Texto Completo: https://www.teses.usp.br/teses/disponiveis/3/3137/tde-02062022-090659/
Resumo: Ethanol production in Brazil occurs through the fermentation of sugarcane juice or molasses by yeasts of the species Saccharomyces cerevisiae. The use of molasses has logistical and economic advantages compared to sugar cane juice, due to its reduced moisture content and high concentration of sugars. However, the quality of the molasses varies even in the same mill, depending on factors such as the quality of the cane, fertilization and the number of recycling to which the molasses was submitted for sugar removal. For this reason, the use of molasses for academic research becomes problematic because it is not possible to guarantee that a given lot of molasses is representative and thus, the reproducibility of experiments is impaired, making its use difficult by research centers. Thus, the objective of this work is to develop a defined medium that is able to reproduce physiological and technological parameters from real molasses. As the first iteration of this medium to be proposed, a published semi-synthetic medium intended to simulate sugar cane molasses must was tested in two industrial lines and two laboratory strains of S. cerevisiae and not able to adequately reproduce the batch growths for the four distinct strains compared to real molasses from sugarcane mills. Thus, the medium was adapted, generating the 1SMol synthetic medium, which had its composition divided into seven nutritional groups that had their effects on the growth of an industrial yeast strain (PE-2) and a laboratory strain (CEN.PK113-7D) in microplate batch growth experiments. The nutritional groups that most impacted the specific growth rate and maximum absorbance were vitamins and the nitrogen source. However, when compared to real molasses, 1SMol presents high specific growth velocity values lower than real molasses. The medium was then adapted, generating 2SMol, a defined medium, with results in simple batches similar to industrial molasses. For this medium, it was observed that the options of vitamins, ammonium and amino acids have had a greater impact on growth parameters in batches. The 2SMol medium was then used in a batch-fed experiment with cell recycling, simulating the Brazilian industrial process of ethanol production, obtaining results similar to the three industrial molasses regarding viability parameters, wine pH, ethanol yield, profile of CO2 and biomass production over five cycles. Thus, the 2SMol medium is presented as a good alternative as a standard medium for carrying out studies involving the fermentation of sugarcane molasses by yeast.
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spelling Development of a defined synthetic medium to mimic sugarcane molasses for ethanol fermentation.Desenvolvimento de um meio de cultura sintético definido para imitar melaço de cana-de-açúcar em fermentação etanólica.EtanolEthanolic fermentationFermentação etanólicaMelaçoMolassesSaccharomycesSaccharomyces cerevisiaeEthanol production in Brazil occurs through the fermentation of sugarcane juice or molasses by yeasts of the species Saccharomyces cerevisiae. The use of molasses has logistical and economic advantages compared to sugar cane juice, due to its reduced moisture content and high concentration of sugars. However, the quality of the molasses varies even in the same mill, depending on factors such as the quality of the cane, fertilization and the number of recycling to which the molasses was submitted for sugar removal. For this reason, the use of molasses for academic research becomes problematic because it is not possible to guarantee that a given lot of molasses is representative and thus, the reproducibility of experiments is impaired, making its use difficult by research centers. Thus, the objective of this work is to develop a defined medium that is able to reproduce physiological and technological parameters from real molasses. As the first iteration of this medium to be proposed, a published semi-synthetic medium intended to simulate sugar cane molasses must was tested in two industrial lines and two laboratory strains of S. cerevisiae and not able to adequately reproduce the batch growths for the four distinct strains compared to real molasses from sugarcane mills. Thus, the medium was adapted, generating the 1SMol synthetic medium, which had its composition divided into seven nutritional groups that had their effects on the growth of an industrial yeast strain (PE-2) and a laboratory strain (CEN.PK113-7D) in microplate batch growth experiments. The nutritional groups that most impacted the specific growth rate and maximum absorbance were vitamins and the nitrogen source. However, when compared to real molasses, 1SMol presents high specific growth velocity values lower than real molasses. The medium was then adapted, generating 2SMol, a defined medium, with results in simple batches similar to industrial molasses. For this medium, it was observed that the options of vitamins, ammonium and amino acids have had a greater impact on growth parameters in batches. The 2SMol medium was then used in a batch-fed experiment with cell recycling, simulating the Brazilian industrial process of ethanol production, obtaining results similar to the three industrial molasses regarding viability parameters, wine pH, ethanol yield, profile of CO2 and biomass production over five cycles. Thus, the 2SMol medium is presented as a good alternative as a standard medium for carrying out studies involving the fermentation of sugarcane molasses by yeast.A produção de etanol no Brasil ocorre através da fermentação do caldo ou melaço de cana-deaçúcar por leveduras da espécie Saccharomyces cerevisiae. O uso do melaço apresenta vantagens logísticas e econômicas em relação ao caldo, devido ao seu teor de umidade reduzido e alta concentração de açúcares. Entretanto, a qualidade dos melaços varia mesmo em uma mesma usina, em função de fatores como a qualidade da cana, fertilização e número de reciclos ao qual o melaço foi submetido para a etapa de cristalização de açúcar. Por isto, utilização de melaços para pesquisas torna-se problemática pois não é possível garantir que um determinado lote de melaço seja adequadamente representativo e assim, a reprodutibilidade de experimentos é prejudicada, dificultando sua utilização em diferentes laboratórios. Desta maneira, o objetivo deste trabalho é desenvolver um meio definido que seja capaz de reproduzir parâmetros fisiológicos e tecnológicos obtidos a partir de amostras de melaços reais. Como primeira iteração deste meio a ser proposto, uma formulação semidefinida recentemente publicada para simular mosto de melaço de cana foi testada em duas linhagens industriais e duas linhagens laboratoriais de S. cerevisiae e esta não foi capaz de reproduzir de maneira adequada os crescimentos em batelada para as quatro linhagens distintas em comparação aos melaços reais de usinas alcooleiras. Assim, o meio foi adaptado, gerando o meio sintético 1SMol, que teve sua composição dividida em sete grupos nutricionais que tiveram seus efeitos avaliados sobre o crescimento de uma levedura industrial (PE-2) e uma laboratorial (CEN.PK113-7D), através de ensaios de crescimento em batelada. Os grupos nutricionais que mais impactaram na velocidade específica de crescimento e na absorbância máxima foram as vitaminas e a fonte de nitrogênio. Entretanto, ao ter os resultados comparados com melaços reais, o meio 1SMol apresentou valores de máxima velocidade específica de crescimento menores que melaços reais. O meio foi então adaptado, gerando 2SMol, um meio definido, com resultados em bateladas simples similares aos de melaços industriais. Para este meio, foi observado que as concentrações de vitaminas, amônio e aminoácidos tiveram maior impacto sobre parâmetros de crescimento em batelada. O meio 2SMol foi então comparado a três melaços industriais em um experimento de batelada alimentada com reciclo de células, simulando o processo industrial brasileiro de produção de etanol. Resultados similares foram obtidos para os cultivos com a formulação sintética e os meios industriais com relação aos parâmetros viabilidade, pH de vinho, rendimento em etanol, perfil de produção de CO2 e biomassa ao longo de cinco reciclos. Assim, o meio 2SMol se apresenta como uma boa alternativa de meio de cultura padronizado para a realização de estudos envolvendo a fermentação de melaço de cana-de-açúcar por leveduras.Biblioteca Digitais de Teses e Dissertações da USPBasso, Thiago OlittaCunha, Gabriel Caetano de Gois e2022-03-11info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttps://www.teses.usp.br/teses/disponiveis/3/3137/tde-02062022-090659/reponame:Biblioteca Digital de Teses e Dissertações da USPinstname:Universidade de São Paulo (USP)instacron:USPLiberar o conteúdo para acesso público.info:eu-repo/semantics/openAccesseng2022-06-02T12:27:22Zoai:teses.usp.br:tde-02062022-090659Biblioteca Digital de Teses e Dissertaçõeshttp://www.teses.usp.br/PUBhttp://www.teses.usp.br/cgi-bin/mtd2br.plvirginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.bropendoar:27212022-06-02T12:27:22Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)false
dc.title.none.fl_str_mv Development of a defined synthetic medium to mimic sugarcane molasses for ethanol fermentation.
Desenvolvimento de um meio de cultura sintético definido para imitar melaço de cana-de-açúcar em fermentação etanólica.
title Development of a defined synthetic medium to mimic sugarcane molasses for ethanol fermentation.
spellingShingle Development of a defined synthetic medium to mimic sugarcane molasses for ethanol fermentation.
Cunha, Gabriel Caetano de Gois e
Etanol
Ethanolic fermentation
Fermentação etanólica
Melaço
Molasses
Saccharomyces
Saccharomyces cerevisiae
title_short Development of a defined synthetic medium to mimic sugarcane molasses for ethanol fermentation.
title_full Development of a defined synthetic medium to mimic sugarcane molasses for ethanol fermentation.
title_fullStr Development of a defined synthetic medium to mimic sugarcane molasses for ethanol fermentation.
title_full_unstemmed Development of a defined synthetic medium to mimic sugarcane molasses for ethanol fermentation.
title_sort Development of a defined synthetic medium to mimic sugarcane molasses for ethanol fermentation.
author Cunha, Gabriel Caetano de Gois e
author_facet Cunha, Gabriel Caetano de Gois e
author_role author
dc.contributor.none.fl_str_mv Basso, Thiago Olitta
dc.contributor.author.fl_str_mv Cunha, Gabriel Caetano de Gois e
dc.subject.por.fl_str_mv Etanol
Ethanolic fermentation
Fermentação etanólica
Melaço
Molasses
Saccharomyces
Saccharomyces cerevisiae
topic Etanol
Ethanolic fermentation
Fermentação etanólica
Melaço
Molasses
Saccharomyces
Saccharomyces cerevisiae
description Ethanol production in Brazil occurs through the fermentation of sugarcane juice or molasses by yeasts of the species Saccharomyces cerevisiae. The use of molasses has logistical and economic advantages compared to sugar cane juice, due to its reduced moisture content and high concentration of sugars. However, the quality of the molasses varies even in the same mill, depending on factors such as the quality of the cane, fertilization and the number of recycling to which the molasses was submitted for sugar removal. For this reason, the use of molasses for academic research becomes problematic because it is not possible to guarantee that a given lot of molasses is representative and thus, the reproducibility of experiments is impaired, making its use difficult by research centers. Thus, the objective of this work is to develop a defined medium that is able to reproduce physiological and technological parameters from real molasses. As the first iteration of this medium to be proposed, a published semi-synthetic medium intended to simulate sugar cane molasses must was tested in two industrial lines and two laboratory strains of S. cerevisiae and not able to adequately reproduce the batch growths for the four distinct strains compared to real molasses from sugarcane mills. Thus, the medium was adapted, generating the 1SMol synthetic medium, which had its composition divided into seven nutritional groups that had their effects on the growth of an industrial yeast strain (PE-2) and a laboratory strain (CEN.PK113-7D) in microplate batch growth experiments. The nutritional groups that most impacted the specific growth rate and maximum absorbance were vitamins and the nitrogen source. However, when compared to real molasses, 1SMol presents high specific growth velocity values lower than real molasses. The medium was then adapted, generating 2SMol, a defined medium, with results in simple batches similar to industrial molasses. For this medium, it was observed that the options of vitamins, ammonium and amino acids have had a greater impact on growth parameters in batches. The 2SMol medium was then used in a batch-fed experiment with cell recycling, simulating the Brazilian industrial process of ethanol production, obtaining results similar to the three industrial molasses regarding viability parameters, wine pH, ethanol yield, profile of CO2 and biomass production over five cycles. Thus, the 2SMol medium is presented as a good alternative as a standard medium for carrying out studies involving the fermentation of sugarcane molasses by yeast.
publishDate 2022
dc.date.none.fl_str_mv 2022-03-11
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