Desenvolvimento e aplicação de métodos analíticos para caracterização de biomassas microalgais
Ano de defesa: | 2020 |
---|---|
Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | , , , |
Tipo de documento: | Tese |
Tipo de acesso: | Acesso aberto |
Idioma: | por |
Instituição de defesa: |
Universidade Federal de Santa Maria
Centro de Ciências Rurais |
Programa de Pós-Graduação: |
Programa de Pós-Graduação em Ciência e Tecnologia dos Alimentos
|
Departamento: |
Ciência e Tecnologia dos Alimentos
|
País: |
Brasil
|
Palavras-chave em Português: | |
Palavras-chave em Inglês: | |
Área do conhecimento CNPq: | |
Link de acesso: | http://repositorio.ufsm.br/handle/1/25293 |
Resumo: | Microalgal biomass has compounds of commercial importance, lipids, proteins, carbohydrates, pigments and toxins are the most researched and applied. However, synthesis intermediates can also be studied to help understand the metabolic pathways performed by these microorganisms. The objective of this work was to evaluate microalgal metabolites and to characterize biomass obtained from cultivations with different external conditions. Analyzes were carried out with integrated sample preparation methods, and for pigments, specifically, a new extraction method was developed. Except for pigments that are thermally unstable and determined by spectrophotometry or liquid chromatography (LC), gas chromatography (GC) presents itself as a suitable analytical tool for characterizing metabolites. This technique has the advantage of separating and detecting gases, volatile organic compounds, polar (organic acids, amino acids, sugars and sugar alcohols) and non-polar (fatty acids and sterols) metabolites, in addition to high resolution and selectivity. GC was used in the study of the Scenedesmus obliquus metabolomics, allowing the identification and quantification of organic acids, free amino acids and fatty acids. In addition to the precursors, proteins, lipids and total chlorophylls were determined. The synthesis of precursor metabolites was affected by the external cultivation conditions, different photoperiods, 24:0 and 12:12 (hour:hour; light:dark), and also by the phases of cell growth, exponential, stationary and declining. At the end of the cultivations, the highest concentrations of biomass (4020 mg L-1) and protein (47.30%) were obtained in the cultivation with constant lighting. In the 12:12 photoperiod (light:dark) culture, S. obliquus triggered a higher production of lipids (23.00%) and chlorophylls (26.40 mg g-1). The major pigments present in microalgae are chlorophylls and carotenoids, and there are still limitations in the extraction methods applied in these biomasses, such as high solvent consumption and time. For the method development of simultaneous extraction of chlorophylls and carotenoids from the microalgae Spirulina sp. and S. obliquus, different mechanical homogenization times and extracting solvents were evaluated. Satisfactory recovery values, from 101.11-124.18% for most pigments, in relation to the maceration method (MM) used as a reference, were obtained with 30 min of homogenization and extraction with chloroform followed by ethanol (PM30E). In general, the PM30E pigment profiles showed fewer isomers and possible degradation compounds, in addition to a reduction of about five times in the volume of organic solvents and is four times faster than MM. Subsequently, the characterization of S. obliquus biomasses cultivated with 3, 5, 10, 15, 20 and 25% of CO2 was carried out. Cultures with 3, 5, and 10% CO2 showed greater synthesis of amino acids and proteins (greater than 60% of dry biomass). The highest concentrations of chlorophylls were found in cultures with 15, 20 and 25% CO2, 24.23, 23.06 and 30.79 mg g-1. The total carotenoids were three times bigger cultures with 3, 5 and 10% of CO2 and all-trans-β-carotene was the main compound. The synthesis of lipids was intensified with the increase of CO2 percentages, as well as the accumulation of polyunsaturated fatty acids. |
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2022-07-08T12:50:05Z2022-07-08T12:50:05Z2020-10-30http://repositorio.ufsm.br/handle/1/25293Microalgal biomass has compounds of commercial importance, lipids, proteins, carbohydrates, pigments and toxins are the most researched and applied. However, synthesis intermediates can also be studied to help understand the metabolic pathways performed by these microorganisms. The objective of this work was to evaluate microalgal metabolites and to characterize biomass obtained from cultivations with different external conditions. Analyzes were carried out with integrated sample preparation methods, and for pigments, specifically, a new extraction method was developed. Except for pigments that are thermally unstable and determined by spectrophotometry or liquid chromatography (LC), gas chromatography (GC) presents itself as a suitable analytical tool for characterizing metabolites. This technique has the advantage of separating and detecting gases, volatile organic compounds, polar (organic acids, amino acids, sugars and sugar alcohols) and non-polar (fatty acids and sterols) metabolites, in addition to high resolution and selectivity. GC was used in the study of the Scenedesmus obliquus metabolomics, allowing the identification and quantification of organic acids, free amino acids and fatty acids. In addition to the precursors, proteins, lipids and total chlorophylls were determined. The synthesis of precursor metabolites was affected by the external cultivation conditions, different photoperiods, 24:0 and 12:12 (hour:hour; light:dark), and also by the phases of cell growth, exponential, stationary and declining. At the end of the cultivations, the highest concentrations of biomass (4020 mg L-1) and protein (47.30%) were obtained in the cultivation with constant lighting. In the 12:12 photoperiod (light:dark) culture, S. obliquus triggered a higher production of lipids (23.00%) and chlorophylls (26.40 mg g-1). The major pigments present in microalgae are chlorophylls and carotenoids, and there are still limitations in the extraction methods applied in these biomasses, such as high solvent consumption and time. For the method development of simultaneous extraction of chlorophylls and carotenoids from the microalgae Spirulina sp. and S. obliquus, different mechanical homogenization times and extracting solvents were evaluated. Satisfactory recovery values, from 101.11-124.18% for most pigments, in relation to the maceration method (MM) used as a reference, were obtained with 30 min of homogenization and extraction with chloroform followed by ethanol (PM30E). In general, the PM30E pigment profiles showed fewer isomers and possible degradation compounds, in addition to a reduction of about five times in the volume of organic solvents and is four times faster than MM. Subsequently, the characterization of S. obliquus biomasses cultivated with 3, 5, 10, 15, 20 and 25% of CO2 was carried out. Cultures with 3, 5, and 10% CO2 showed greater synthesis of amino acids and proteins (greater than 60% of dry biomass). The highest concentrations of chlorophylls were found in cultures with 15, 20 and 25% CO2, 24.23, 23.06 and 30.79 mg g-1. The total carotenoids were three times bigger cultures with 3, 5 and 10% of CO2 and all-trans-β-carotene was the main compound. The synthesis of lipids was intensified with the increase of CO2 percentages, as well as the accumulation of polyunsaturated fatty acids.A biomassa microalgal apresenta compostos de importância comercial, lipídios, proteínas, carboidratos, pigmentos e toxinas são os mais pesquisados e aplicados. Entretanto, intermediários da síntese também podem ser estudados, para auxiliar no entendimento das vias metabólicas desempenhadas por esses microrganismos. O objetivo desse trabalho foi avaliar metabólitos microalgais e caracterizar biomassas obtidas a partir de cultivos com diferentes condições externas. As análises foram realizadas com métodos integrados de preparo de amostra e para pigmentos, especificamente, foi desenvolvido um novo método de extração. Com exceção dos pigmentos que são termo instáveis e determinados por espectrofotometria ou cromatografia líquida (LC), a cromatografia em fase gasosa (GC) se apresenta como uma ferramenta analítica adequada para caracterização de metabólitos. Essa técnica tem como vantagens separação e detecção de gases, compostos orgânicos voláteis, metabólitos polares (ácidos orgânicos, aminoácidos, açúcares e açúcares álcoois) e não polares (ácidos graxos e esteróis), além de elevada resolução e seletividade. A GC foi emprega no estudo da metabolômica de Scenedesmus obliquus, permitindo a identificação e quantificação de ácidos orgânicos, aminoácidos livres e ácidos graxos. Além dos precursores, foram determinados proteínas, lipídios e clorofilas totais. A síntese dos metabólitos precursores foi afetada pelas condições externas dos cultivos, diferentes fotoperíodos, 24:0 e 12:12 (hora:hora; claro:escuro), e também pelas fases do crescimento celular, exponencial, estacionária e de declínio. Ao final dos cultivos as maiores concentrações de biomassa (4020 mg L-1) e proteína (47,30%) foram obtidas no cultivo com iluminação constante. Já no cultivo com fotoperíodo 12:12 (claro:escuro) a S. obliquus desencadeou maior produção de lipídios (23,00%) e clorofilas (26,4 mg g-1). Os pigmentos majoritários presentes em microalgas são as clorofilas e os carotenoides e ainda existem limitações nos métodos de extração aplicados nessas biomassas, como elevado consumo de solventes e tempo. Para o desenvolvimento do método de extração simultânea de clorofilas e carotenoides das microalgas Spirulina sp. e S. obliquus, foram avaliados diferentes tempos de homogeneização mecânica e solventes extratores. Valores de recuperação satisfatórios, de 101,11-124,18% para maioria dos pigmentos, em relação método de maceração (MM) utilizado como referência, foram obtidos com 30 min de homogeneização e extração com clorofórmio seguida por etanol (PM30E). Em geral, os perfis dos pigmentos do PM30E apresentaram menos isômeros e possíveis compostos de degradação, além da redução de cerca de cinco vezes no volume de solventes orgânicos e ser quatro vezes mais rápido que o MM. Posteriormente, foi realizada a caracterização de biomassas de S. obliquus cultivadas com 3, 5, 10, 15, 20 e 25% de CO2. Cultivos com 3, 5 e 10% de CO2 apresentaram maior síntese de aminoácidos e proteínas (superior a 60% da biomassa seca). As maiores concentrações de clorofilas foram encontradas nos cultivos com 15, 20 e 25% CO2, 24,23, 23,06 e 30,79 mg g-1. Os carotenoides totais foram três vezes maiores cultivos com 3, 5 e 10% de CO2 e all-trans-β-caroteno foi o composto principal. A síntese de lipídeos foi intensificada com o aumento das porcentagens de CO2, assim como o acúmulo de ácidos graxos poli-insaturados.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESporUniversidade Federal de Santa MariaCentro de Ciências RuraisPrograma de Pós-Graduação em Ciência e Tecnologia dos AlimentosUFSMBrasilCiência e Tecnologia dos AlimentosAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessScenedesmus obliquusSpirulina sp.Cromatografia em fase gasosaÁcidos orgânicosAminoácidosÁcidos graxosProteínasLipídiosPigmentosGas chromatographyOrganic acidsAmino acidsFatty acidsProteinsLipidsPigmentsCNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOSDesenvolvimento e aplicação de métodos analíticos para caracterização de biomassas microalgaisDevelopment and application of analytical methods for microalgal biomasses characterizationinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisWagner, Rogerhttp://lattes.cnpq.br/4780821244553957Meinhart, Adriana DillenburgBallus, Cristiano AugustoLopes, Eduardo JacobFurlan, Valcenir Júnior Mendeshttp://lattes.cnpq.br/2894625411934949Vendruscolo, Raquel Guidetti50070000000660060060060060060060097d863bb-f506-4732-abde-1f459447c5664b3af928-98b3-40f6-8f6a-e8982942f74e708b8997-635c-4b35-91f3-79723a9dbd838aed6c33-9c7e-40d6-9fe2-fd5138d7021947c186e2-507d-4040-8e21-0c5b55eb24c12af30eb6-58d5-43d9-881b-71e767dc5befreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMORIGINALTES_PPGCTA_2020_VENDRUSCOLO_RAQUEL.pdfTES_PPGCTA_2020_VENDRUSCOLO_RAQUEL.pdfTese de doutoradoapplication/pdf3583084http://repositorio.ufsm.br/bitstream/1/25293/1/TES_PPGCTA_2020_VENDRUSCOLO_RAQUEL.pdf5a17638fe575a225f7cffc04d44fc4f7MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; 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dc.title.por.fl_str_mv |
Desenvolvimento e aplicação de métodos analíticos para caracterização de biomassas microalgais |
dc.title.alternative.eng.fl_str_mv |
Development and application of analytical methods for microalgal biomasses characterization |
title |
Desenvolvimento e aplicação de métodos analíticos para caracterização de biomassas microalgais |
spellingShingle |
Desenvolvimento e aplicação de métodos analíticos para caracterização de biomassas microalgais Vendruscolo, Raquel Guidetti Scenedesmus obliquus Spirulina sp. Cromatografia em fase gasosa Ácidos orgânicos Aminoácidos Ácidos graxos Proteínas Lipídios Pigmentos Gas chromatography Organic acids Amino acids Fatty acids Proteins Lipids Pigments CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS |
title_short |
Desenvolvimento e aplicação de métodos analíticos para caracterização de biomassas microalgais |
title_full |
Desenvolvimento e aplicação de métodos analíticos para caracterização de biomassas microalgais |
title_fullStr |
Desenvolvimento e aplicação de métodos analíticos para caracterização de biomassas microalgais |
title_full_unstemmed |
Desenvolvimento e aplicação de métodos analíticos para caracterização de biomassas microalgais |
title_sort |
Desenvolvimento e aplicação de métodos analíticos para caracterização de biomassas microalgais |
author |
Vendruscolo, Raquel Guidetti |
author_facet |
Vendruscolo, Raquel Guidetti |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Wagner, Roger |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/4780821244553957 |
dc.contributor.referee1.fl_str_mv |
Meinhart, Adriana Dillenburg |
dc.contributor.referee2.fl_str_mv |
Ballus, Cristiano Augusto |
dc.contributor.referee3.fl_str_mv |
Lopes, Eduardo Jacob |
dc.contributor.referee4.fl_str_mv |
Furlan, Valcenir Júnior Mendes |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/2894625411934949 |
dc.contributor.author.fl_str_mv |
Vendruscolo, Raquel Guidetti |
contributor_str_mv |
Wagner, Roger Meinhart, Adriana Dillenburg Ballus, Cristiano Augusto Lopes, Eduardo Jacob Furlan, Valcenir Júnior Mendes |
dc.subject.por.fl_str_mv |
Scenedesmus obliquus Spirulina sp. Cromatografia em fase gasosa Ácidos orgânicos Aminoácidos Ácidos graxos Proteínas Lipídios Pigmentos |
topic |
Scenedesmus obliquus Spirulina sp. Cromatografia em fase gasosa Ácidos orgânicos Aminoácidos Ácidos graxos Proteínas Lipídios Pigmentos Gas chromatography Organic acids Amino acids Fatty acids Proteins Lipids Pigments CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS |
dc.subject.eng.fl_str_mv |
Gas chromatography Organic acids Amino acids Fatty acids Proteins Lipids Pigments |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::CIENCIA E TECNOLOGIA DE ALIMENTOS |
description |
Microalgal biomass has compounds of commercial importance, lipids, proteins, carbohydrates, pigments and toxins are the most researched and applied. However, synthesis intermediates can also be studied to help understand the metabolic pathways performed by these microorganisms. The objective of this work was to evaluate microalgal metabolites and to characterize biomass obtained from cultivations with different external conditions. Analyzes were carried out with integrated sample preparation methods, and for pigments, specifically, a new extraction method was developed. Except for pigments that are thermally unstable and determined by spectrophotometry or liquid chromatography (LC), gas chromatography (GC) presents itself as a suitable analytical tool for characterizing metabolites. This technique has the advantage of separating and detecting gases, volatile organic compounds, polar (organic acids, amino acids, sugars and sugar alcohols) and non-polar (fatty acids and sterols) metabolites, in addition to high resolution and selectivity. GC was used in the study of the Scenedesmus obliquus metabolomics, allowing the identification and quantification of organic acids, free amino acids and fatty acids. In addition to the precursors, proteins, lipids and total chlorophylls were determined. The synthesis of precursor metabolites was affected by the external cultivation conditions, different photoperiods, 24:0 and 12:12 (hour:hour; light:dark), and also by the phases of cell growth, exponential, stationary and declining. At the end of the cultivations, the highest concentrations of biomass (4020 mg L-1) and protein (47.30%) were obtained in the cultivation with constant lighting. In the 12:12 photoperiod (light:dark) culture, S. obliquus triggered a higher production of lipids (23.00%) and chlorophylls (26.40 mg g-1). The major pigments present in microalgae are chlorophylls and carotenoids, and there are still limitations in the extraction methods applied in these biomasses, such as high solvent consumption and time. For the method development of simultaneous extraction of chlorophylls and carotenoids from the microalgae Spirulina sp. and S. obliquus, different mechanical homogenization times and extracting solvents were evaluated. Satisfactory recovery values, from 101.11-124.18% for most pigments, in relation to the maceration method (MM) used as a reference, were obtained with 30 min of homogenization and extraction with chloroform followed by ethanol (PM30E). In general, the PM30E pigment profiles showed fewer isomers and possible degradation compounds, in addition to a reduction of about five times in the volume of organic solvents and is four times faster than MM. Subsequently, the characterization of S. obliquus biomasses cultivated with 3, 5, 10, 15, 20 and 25% of CO2 was carried out. Cultures with 3, 5, and 10% CO2 showed greater synthesis of amino acids and proteins (greater than 60% of dry biomass). The highest concentrations of chlorophylls were found in cultures with 15, 20 and 25% CO2, 24.23, 23.06 and 30.79 mg g-1. The total carotenoids were three times bigger cultures with 3, 5 and 10% of CO2 and all-trans-β-carotene was the main compound. The synthesis of lipids was intensified with the increase of CO2 percentages, as well as the accumulation of polyunsaturated fatty acids. |
publishDate |
2020 |
dc.date.issued.fl_str_mv |
2020-10-30 |
dc.date.accessioned.fl_str_mv |
2022-07-08T12:50:05Z |
dc.date.available.fl_str_mv |
2022-07-08T12:50:05Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufsm.br/handle/1/25293 |
url |
http://repositorio.ufsm.br/handle/1/25293 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.cnpq.fl_str_mv |
500700000006 |
dc.relation.confidence.fl_str_mv |
600 600 600 600 600 600 600 |
dc.relation.authority.fl_str_mv |
97d863bb-f506-4732-abde-1f459447c566 4b3af928-98b3-40f6-8f6a-e8982942f74e 708b8997-635c-4b35-91f3-79723a9dbd83 8aed6c33-9c7e-40d6-9fe2-fd5138d70219 47c186e2-507d-4040-8e21-0c5b55eb24c1 2af30eb6-58d5-43d9-881b-71e767dc5bef |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências Rurais |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Ciência e Tecnologia dos Alimentos |
dc.publisher.initials.fl_str_mv |
UFSM |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
Ciência e Tecnologia dos Alimentos |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências Rurais |
dc.source.none.fl_str_mv |
reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
instname_str |
Universidade Federal de Santa Maria (UFSM) |
instacron_str |
UFSM |
institution |
UFSM |
reponame_str |
Manancial - Repositório Digital da UFSM |
collection |
Manancial - Repositório Digital da UFSM |
bitstream.url.fl_str_mv |
http://repositorio.ufsm.br/bitstream/1/25293/1/TES_PPGCTA_2020_VENDRUSCOLO_RAQUEL.pdf http://repositorio.ufsm.br/bitstream/1/25293/2/license_rdf http://repositorio.ufsm.br/bitstream/1/25293/3/license.txt |
bitstream.checksum.fl_str_mv |
5a17638fe575a225f7cffc04d44fc4f7 4460e5956bc1d1639be9ae6146a50347 2f0571ecee68693bd5cd3f17c1e075df |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 |
repository.name.fl_str_mv |
Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
|
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1794524287636340736 |