Prospecção bioquímica da biomassa global da cianobactéria tóxica Microcystis aeruginosa BB005
Cyanobacteria are microorganisms able to produce substances with high biological and biotechnological potential. These substances may cause disorders because of the toxins or may be composed by beneficial substances such as carotenoids, lipids and enzymes of technological interest. Hence, the scope...
| Autor principal: | Copelli, Thalita da Silva |
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| Formato: | Dissertação |
| Idioma: | Português |
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Universidade Tecnológica Federal do Paraná
2015
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riut-1-13572015-09-11T06:00:40Z Prospecção bioquímica da biomassa global da cianobactéria tóxica Microcystis aeruginosa BB005 Copelli, Thalita da Silva Pagioro, Thomaz Aurélio Grzybowski, Adelia Coral, Lucila Adriani Ácidos graxos Esteres Carotenóides Fosfatase alcalina Flotação Separação (Tecnologia) Toxicologia ambiental Tecnologia ambiental Fatty acids Esters Carotenoids Alkaline phosphatase Flotation Separation (Technology) Environmental toxicology Green technology Cyanobacteria are microorganisms able to produce substances with high biological and biotechnological potential. These substances may cause disorders because of the toxins or may be composed by beneficial substances such as carotenoids, lipids and enzymes of technological interest. Hence, the scope of the current research was centered in the toxicity evaluation or prospect the substances from the Microcystis aeruginosa cyanobacterium biomass – BB 005. Daphnia magna was used in the toxicity bioassays and the cyanobacterium dry biomass was characterized by the transesterified lipids analysis, carotenoid determination, electrophoretic profile for proteins and acid hydrolysis of polysaccharides. The foam fractionation methodology was performed to separate the phycocyanin pigment and enzymes (e.g. alkaline phosphatase and lipase), using the supernatant from a liquid culture of the cyanobacterium. Although it was possible to quantify the production of toxins by BB005 strain, the ecotoxicological tests have no establish a correlation between toxin concentration and the immobility of organism bodies.The methyl esters profile in which there was a large amount of PUFAs (ω-fatty acids) with 25% of linolenates. The carotenoid content found during the lipids transesterificaction, expressed as β – carotene, was 6.6 mg/g of dry mass. The electrophoresis indicated at least 15 distinct bands of proteins and the chromatographic profile from the native polymeric carbohydrates indicated higher concentration of monosaccharide glucose and a glucose disaccharide, possible maltose. Proteins separation through foaming showed a direct correlation between the nitrogen flow and the collected foam. The phycocianin enrichment factor was high, 41.69, at a 20 mL/min N 2 flow. While in the case of alkaline phosphatase the best result was attained in the 120 mL/min of N2 flow which lead to enrichment and purification factors equal to 5.25 and 2.85 respectively. Therefore, this research has demonstrated the importance of the investigation on compounds from the M. aeruginosa entire biomass, especially the methyl esters from less common and highly insaturated fatty acids, an expressive content of carotenoids and, furthermore, the separation of phycobillin and key- enzymes for biochemical processes, that were feasibly and primarily demonstrated in this experiment by the use of foaming procedure. Cianobactérias são microrganismos capazes de produzir substâncias com potencial biológico e biotecnológico. Tais substâncias podem acarretar em malefícios, causados pelas toxinas, ou ter em sua composição substâncias benéficas como carotenoides, lipídeos e enzimas, de interesse biotecnológico. Assim, esse trabalho teve como objetivo avaliar a toxicidade e prospectar componentes presentes na biomassa global da cianobactéria Microcystis aeruginosa BB005. A toxicidade foi avaliada através de bioensaios com Daphnia magna, e a biomassa seca foi caracterizada por análises dos ésteres metílicos de ácidos graxos (EMAGs), determinação de carotenoides, obtenção do perfil proteico por eletroforese e hidrólise ácida de carboidratos. Também foi empregada uma metodologia de separação adsortiva por bolhas a fim de separar o pigmento ficocianina e enzimas de interesse biotecnológico (e.g. fosfatase alcalina e lipase) utilizando o sobrenadante de um cultivo. Apesar de ter sido possível quantificar a produção de toxinas pela cepa BB005, os ensaios ecotoxicológicos não permitiram estabelecer uma correlação entre a concentração de toxina e a imobilidade dos organismos. O perfil de EMAGs apresentou preponderância de PUFAs (ω-ácidos graxos) sendo que os linolenatos corresponderam a 25% do total. O teor de carotenoides encontrado durante a purificação dos lipídeos transesterificados e expresso como β-caroteno foi de 6,66 mg/g de biomassa seca. A eletroforese mostrou pelo menos 15 bandas distintas de proteínas e o perfil cromatográfico dos hidrolisados de carboidratos mostrou maior concentração do monossacarídeo glucose e de um dissacarídeo de glucose, possivelmente maltose. A separação de proteínas por espumação apresentou uma correlação direta entre fluxo de nitrogênio e a fração de espuma coletada. O fator de enriquecimento para a ficocianina foi de 41,69 utilizando fluxo de 20 mL/min de N2. Para a enzima fosfatase alcalina, os melhores resultados foram obtidos para o fluxo de 120 mL/min de N2, que apresentou fator de enriquecimento e de purificação de 5,25 e 2,85, respectivamente. Portanto, este trabalho demonstrou a importância de investigações sobre os componentes oriundos de M. aeruginosa, sobretudo dos EMAGs incomuns, a quantidade expressiva de carotenos e ainda da separação da ficobilina e de enzimas chave nos processos bioquímicos utilizando o processo por espumação. 5000 2015-09-10T17:29:33Z 2015-02-12 masterThesis COPELLI, Thalita da Silva. Prospecção bioquímica da biomassa global da cianobactéria tóxica Microcystis aeruginosa BB005. 2015. 109 f. Dissertação (Mestrado em Engenharia Tecnologia Ambiental) – Universidade Tecnológica Federal do Paraná, Curitiba, 2015. http://repositorio.utfpr.edu.br/jspui/handle/1/1357 por application/pdf Universidade Tecnológica Federal do Paraná Curitiba Programa de Pós-Graduação em Ciência e Tecnologia Ambiental |
| institution |
Universidade Tecnológica Federal do Paraná |
| collection |
RIUT |
| language |
Português |
| topic |
Ácidos graxos Esteres Carotenóides Fosfatase alcalina Flotação Separação (Tecnologia) Toxicologia ambiental Tecnologia ambiental Fatty acids Esters Carotenoids Alkaline phosphatase Flotation Separation (Technology) Environmental toxicology Green technology |
| spellingShingle |
Ácidos graxos Esteres Carotenóides Fosfatase alcalina Flotação Separação (Tecnologia) Toxicologia ambiental Tecnologia ambiental Fatty acids Esters Carotenoids Alkaline phosphatase Flotation Separation (Technology) Environmental toxicology Green technology Copelli, Thalita da Silva Prospecção bioquímica da biomassa global da cianobactéria tóxica Microcystis aeruginosa BB005 |
| description |
Cyanobacteria are microorganisms able to produce substances with high biological and biotechnological potential. These substances may cause disorders because of the toxins or may be composed by beneficial substances such as carotenoids, lipids and enzymes of technological interest. Hence, the scope of the current research was centered in the toxicity evaluation or prospect the substances from the Microcystis aeruginosa cyanobacterium biomass – BB 005. Daphnia magna was used in the toxicity bioassays and the cyanobacterium dry biomass was characterized by the transesterified lipids analysis, carotenoid determination, electrophoretic profile for proteins and acid hydrolysis of polysaccharides. The foam fractionation methodology was performed to separate the phycocyanin pigment and enzymes (e.g. alkaline phosphatase and lipase), using the supernatant from a liquid culture of the cyanobacterium. Although it was possible to quantify the production of toxins by BB005 strain, the ecotoxicological tests have no establish a correlation between toxin concentration and the immobility of organism bodies.The methyl esters profile in which there was a large amount of PUFAs (ω-fatty acids) with 25% of linolenates. The carotenoid content found during the lipids transesterificaction, expressed as β – carotene, was 6.6 mg/g of dry mass. The electrophoresis indicated at least 15 distinct bands of proteins and the chromatographic profile from the native polymeric carbohydrates indicated higher concentration of monosaccharide glucose and a glucose disaccharide, possible maltose. Proteins separation through foaming showed a direct correlation between the nitrogen flow and the collected foam. The phycocianin enrichment factor was high, 41.69, at a 20 mL/min N 2 flow. While in the case of alkaline phosphatase the best result was attained in the 120 mL/min of N2 flow which lead to enrichment and purification factors equal to 5.25 and 2.85 respectively. Therefore, this research has demonstrated the importance of the investigation on compounds from the M. aeruginosa entire biomass, especially the methyl esters from less common and highly insaturated fatty acids, an expressive content of carotenoids and, furthermore, the separation of phycobillin and key- enzymes for biochemical processes, that were feasibly and primarily demonstrated in this experiment by the use of foaming procedure. |
| format |
Dissertação |
| author |
Copelli, Thalita da Silva |
| author_sort |
Copelli, Thalita da Silva |
| title |
Prospecção bioquímica da biomassa global da cianobactéria tóxica Microcystis aeruginosa BB005 |
| title_short |
Prospecção bioquímica da biomassa global da cianobactéria tóxica Microcystis aeruginosa BB005 |
| title_full |
Prospecção bioquímica da biomassa global da cianobactéria tóxica Microcystis aeruginosa BB005 |
| title_fullStr |
Prospecção bioquímica da biomassa global da cianobactéria tóxica Microcystis aeruginosa BB005 |
| title_full_unstemmed |
Prospecção bioquímica da biomassa global da cianobactéria tóxica Microcystis aeruginosa BB005 |
| title_sort |
prospecção bioquímica da biomassa global da cianobactéria tóxica microcystis aeruginosa bb005 |
| publisher |
Universidade Tecnológica Federal do Paraná |
| publishDate |
2015 |
| citation |
COPELLI, Thalita da Silva. Prospecção bioquímica da biomassa global da cianobactéria tóxica Microcystis aeruginosa BB005. 2015. 109 f. Dissertação (Mestrado em Engenharia Tecnologia Ambiental) – Universidade Tecnológica Federal do Paraná, Curitiba, 2015. |
| url |
http://repositorio.utfpr.edu.br/jspui/handle/1/1357 |
| _version_ |
1805296738260484096 |
| score |
10,814766 |