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Master's Dissertation
DOI
10.11606/D.42.2013.tde-07102013-093554
Document
Author
Full name
Liege Abdallah Kawai
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2013
Supervisor
Committee
Gomez, José Gregorio Cabrera (President)
Baldini, Regina Lúcia
Contiero, Jonas
Title in Portuguese
Melhoramento da eficiência de produção de polihidroxialcanoatos por Pseudomonas sp. através da análise molecular e modificação genética.
Keywords in Portuguese
Pseudomonas
Glicose
Metabolismo de carboidrato
Abstract in Portuguese
A análise de fluxos metabólicos para a produção de PHAMCL por Pseudomonas sp. LFM046 revelou que a baixa eficiência de conversão de carboidratos em PHA (60-70% do valor máximo teórico) deve estar associada à utilização principal da via das pentoses (VP) para o metabolismo de carboidratos. Eficiências significativamente maiores poderiam ser obtidas se uma parcela maior da glicose fosse metabolizada pela via Entner-Doudoroff (ED). Assim, neste trabalho foi realizada a análise de fluxos metabólicos utilizando glicose marcada (C13) e o melhoramento genético de Pseudomonas sp. LFM046 utilizando estratégia de engenharia metabólica. Experimentos com glicose marcada confirmaram um maior fluxo de carboidratos pela VP em relação a ED. A superexpressão de genes específicos de ED em Pseudomonas sp. LFM046 demonstrou apenas pequenos aumentos na eficiência de conversão de carboidratos em PHA pelas linhagens recombinantes.
Title in English
Improvement of the efficiency of polyhydroxyalkanoates production by Pseudomonas sp. through molecular analysis and genetic modification.
Keywords in English
Pseudomonas
Glucose
Metabolism of carbohydrates
Abstract in English
The metabolic flux analysis for PHAMCL production by Pseudomonas sp. LFM046 revealed that the low efficiency of carbohydrates conversion into PHA (60-70% of the maximum theoretical value) should be associated with the main use of pentose phosphate pathway (PP) for the carbohydrate metabolism. Significantly higher efficiencies could be obtained if a larger portion of the glucose was metabolized via the Entner-Doudoroff (ED). Therefore, the metabolic fluxes analysis was performed in this work using labeled glucose (13C) and the genetic improvement of Pseudomonas sp. LFM046 by using metabolic engineering approach. Experiments with labeled glucose confirmed an increased flow of carbohydrates by PP compared to ED. The overexpression of specific genes from ED in Pseudomonas sp. LFM046 showed only small increases in the efficiency of carbohydrates conversion into PHA by recombinant strains.
 
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Publishing Date
2013-12-06
 
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