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Doctoral Thesis
DOI
https://doi.org/10.11606/T.87.2014.tde-27012015-105824
Document
Author
Full name
Thatiane Teixeira Mendonça
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2014
Supervisor
Committee
Silva, Luiziana Ferreira da (President)
Araujo, Welington Luiz de
Barbosa, Heloiza Ramos
Carvalho, Joao Carlos Monteiro de
Piccoli, Rosane Aparecida Moniz
Title in Portuguese
Estudo de bactérias recombinantes e análise de fluxos metabólicos para biossíntese do copolímero biodegrádavel poli(3-hidroxibutirato-co-3-hidroxihexanoato) [P(3HB-co-3HHx).
Keywords in Portuguese
Burkholderia sacchari
Análise de fluxos metabólicos
Copolímeros biodegradáveis
Operon phaPCJ Aeromonas spp.
Poli(3-hidroxibutirato-co-3-hidroxihexanoato)
Recombinantes
Abstract in Portuguese
O copolímero biodegradável poli(3-hidroxibutirato-co-3-hidroxihexanoato) P(3HB-co-3HHx) é um polihidroxialcanoato (PHA) que apresenta várias aplicações. A bactéria Burkholderia sacchari acumula P(3HB-co-2mol%3HHx), a partir de glicose e ácido hexanoico. Com o objetivo de obter P(3HB-co-3HHx) com diferentes teores de 3HHx por B. sacchari, foram construídas linhagens recombinantes, contendo genes do operon phaPCJ de Aeromonas spp. Os recombinantes produziram P(3HB-co-3HHx), a partir de ácidos hexanoico, láurico e linoleico, com teores de 3HHx entre 1,88-18 mol%. Experimentos em biorreator com o recombinante, alimentada na fase de acúmulo por glicose 140 g/L e ácido hexanoico entre 0-45 g/L, resultaram copolímeros com composições variando de 0 a 20 mol% de 3HHx. Os copolímeros assim produzidos foram extraídos e analisados quanto às propriedades físicas. A análise de fluxos metabólicos indicou que a produção de PHA pode ser aumentada com mudanças no metabolismo central e deleção/superexpressão de genes.
Title in English
Study of recombinant bacteria and metabolic flux analysis to biosynthesize the biodegradable copolymer poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)].
Keywords in English
Burkholderia sacchari
phaPCJ operon from Aeromonas spp
Biodegradable copolymers
Metabolic flux analysis.
Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)
Recombinant
Abstract in English
The biodegradable copolymer poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) P(3HB-co-3HHx) is a polyhydroxyalkanoate (PHA) presenting various applications. The bacterium Burkholderia sacchari accumulated P(3HB-co-2mol%3HHx) from glucose and hexanoic acid. In order to obtain P(3HB-co-3HHx) with different 3HHx amounts by B. sacchari, recombinant strains containing phaPCJ operon genes from Aeromonas spp were constructed. Recombinant strains produced P(3HB-co-3HHx) from hexanoic, lauric and linoleic acids, with contents of 3HHx ranging from 1.88 to 18 mol%. Experiments with the recombinant in bioreactor, fed in the accumulation phase by glucose 140 g.l-1and hexanoic acid 0-45 g.l-1, resulted in copolymers with compositions ranging from 0 to 20 mol% of 3HHx. The copolymers produced were extracted and analyzed for physical properties. The metabolic flux analysis indicated that PHA production can be increased by modifying the central metabolism and deleting/ overexpressing genes.
 
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Publishing Date
2015-01-28
 
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