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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2015.tde-04092015-133523
Document
Author
Full name
Carlos Andrés Fajardo Gómez
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2015
Supervisor
Committee
Silva, Luiziana Ferreira da (President)
Maldonado, Gabriel Padilla
Mendonça, Thatiane Teixeira
Title in Portuguese
Desenvolvimento de processo de produção de polihidroxibutirato a partir da xilose empregando técnicas de engenharia evolutiva e bioprocessos.
Keywords in Portuguese
Burkholderia sachhari
Análise de fluxos metabólicos
Metabolismo de xilose
Polihidroxialcanoatos (PHA)
Abstract in Portuguese
O trabalho é proposto visando melhorar o consumo de xilose na bactéria Burkholderia sacchari utilizando o acúmulo de PHB como modelo de produção Foi desenvolvido um processo de evolução por meio da aplicação de feast and famine e Cultivos sequenciais em fase exponencial. Foi obtida uma linhagem mutante com uma velocidade especifica de crescimento de 0,24 h -1. Foi feita uma análise de fluxos metabólicos da qual foi possível concluir que o metabolismo da xilose acontecia em sua maioria pela VP junto com a ED. Foi feito um ensaio de acumulo com carbono marcado utilizando uma solução de xilose, de 20:80 de xilose marcada 13C em todos os carbonos e xilose não marcado, para determinar quais seriam as possíveis vias metabólicas no uso da xilose por parte de B. sacharia LFM 101 e da linhagem evoluída BSEV11. Foi determinado que houve embaralhamento de carbonos, fato que só acontece quando o metabolismo da xilose e feito pela VP junto com a via ED, assim foi possível conferir a via ED como principal via para o metabolismo da xilose em B. sacchari LFM 101.
Title in English
Development of polyhydroxybutyrate production from xylose employing evolutionary engineering techniques and bioprocesses.
Keywords in English
Burkholderia sacchari
Metabolic flux analysis
Polyhydroxyalkanoates (PHA)
Xylose metabolism
Abstract in English
To evaluate the possibilities of improving the productivity of PHA production from xylose, evolutionary engineering techniques were applied to B. sacchari to select cells with maximum specific growth rates (max) higher than the wild type. Metabolic flux analysis was also performed to evaluate the fluxes through central pathways and the possibility of further improvements by modifying fluxes rates. The evolved strain reached a max of 0.24 ± 0.01 h-1 at the end of the evolutionary process. Strains were submitted to bioreactor experiments. A metabolic network of the strain was usedn to determine the possible distribution of metabolic fluxes. A total of 19 elementary modes were obtained. It was concluded that the metabolism of xylose occurred mostly by VP along with the ED. The ED pathway has the major activity going on in a cyclic way. It was also performed a 13C labeled xylose assay, in which it was possibly to confirm the obtained results from the metabolic flux analyses.
 
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Publishing Date
2015-09-04
 
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