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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2013.tde-13052014-164703
Document
Author
Full name
Débora Vieira Parrine Sant'Ana
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2013
Supervisor
Committee
Silva, Luiziana Ferreira da (President)
Pessoa Junior, Adalberto
Spira, Beny
Title in Portuguese
Análise de fluxos metabólicos aplicada à biossíntese do polímero biodegradável poli-3-hidroxi-butirato P(3HB) por Burkholderia sacchari.
Keywords in Portuguese
Bactérias gram-negativas
Carbono
Metabolismo
Pseudomonas
Reatores químicos
Abstract in Portuguese
Este trabalho utiliza a Análise de Fluxos Metabólicos para estudar o aumento da eficiência da linhagem Burkholderia sacchari (LFM101) na produção de PHB. Foram avaliadas as eficiências de conversão de açúcares em PHA de LFM101. Esta também foi cultivada em batelada alimentada em reator, apresentando o máximo teórico durante um estado pseudo-estacionário sob oferta de glicose. Estes dados, submetidos ao software Metatool, resultaram em mapa metabólico contendo os fluxos das reações centrais ede PHA ocorrido no experimento. Através do cultivo de LFM101 e C. necator sob oferta de glicose marcada com 13C, determinou-se que estas utilizam as mesmas vias para produção de PHA, não justificando a baixa eficiência de LFM101. Em um projeto paralelo, estudou-se a eficiência da produção de PHB utilizando melaço de cana, glicerol cru e acetato, em produtores de hidrogênio e PHA, onde verificou-se não apenas o aumento de PHA em mutantes nifh- de R. capsulatus mas a interação dos parâmetros luz e nitrogênio a partir das metodologias DOE e RSM.
Title in English
Metabolic flux analysis applied to the biosynthesis of the biodegradable polymer poly-3-hydroxybutyrate (P3HB) produced by Burkholderia sacchari.
Keywords in English
Carbon
Chemical reactors
Gram-negative bacteria
Metabolism
Pseudomonas
Abstract in English
This work applies Metabolic Flux Analysis to discuss the eficiency of Burkholderia sacchari (LFM101) in PHA production from sugars . Conversion yields of LFM101 and Cupriavidus necator from carbohydrates to PHA were assessed. LFM101 was also grown in reactor fed-batch cultivations, and presented the theoretical maximum in a pseudo-steady stage while grown on glucose. These data were submitted to Metatool and resulted in a metabolic network containing the experimental fluxes of central and PHA metabolism. Cultivation of LFM101 and C. necator under 13C- labeled glucose showed that both species use the same metabolic pathways for the biodegradable polymer synthesis. On a parallel project, the efficiency of biopolymer production from molasses, raw glycerol and acetate in strains producing hydrogen and PHA was tested. Results showed that there was not only an increase in PHA production by R. capsulatus nifH- mutants but also the interaction of light and nitrogen effects were studied by DOE and RSM.
 
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Publishing Date
2014-05-20
 
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