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Doctoral Thesis
DOI
https://doi.org/10.11606/T.87.2018.tde-16032016-101520
Document
Author
Full name
Amanda Billia Flora
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2015
Supervisor
Committee
Gomez, José Gregorio Cabrera (President)
Barros, Mário Henrique de
Galhardo, Rodrigo da Silva
Maldonado, Gabriel Padilla
Marques, Marilis do Valle
Title in Portuguese
Detecção e clonagem de genes de biossíntes de 1,3-propanodiol a partir de glicerol em Klebisiella pneumoniae GLC29.
Keywords in Portuguese
1,3-propanodiol
Klebsiella pneumoniae
Análise de fluxos metabólicos
Biodiesel
Biologia molecular
Glicerol
Isocitrato desidrogenase NAD dependente
Abstract in Portuguese
O 1,3-propanodiol é um composto produzido a partir da fermentação do glicerol por bactérias como a do gênero Klebsiella e que pode ser utilizado para diversas aplicações na indústria, como a produção de PTT (politrimetileno tereftalato). Esse trabalho focou na produção de 1,3-propanodiol utilizando os genes que codificam as enzimas da via retirados de uma cepa de Klebsiella previamente isolada (GLC29). Os genes foram inseridos em um plasmídeo e este em cepa hospedeira de Escherichia coli com alteração na afinidade da enzima isocitrato desidrogenase pelo seu cofator. A enzima foi modificada no genoma da bactéria e utiliza como cofator NAD em vez do NADP original. Ensaios preliminares em agitador rotativo mostraram uma melhoria de produção da cepa mutada em comparação com a Escherichia coli selvagem. Em ensaios em biorreator, variando a concentração de oxigênio dissolvido no meio, a cepa mutante apresentou uma produção 25% maior que a cepa selvagem na condição de maior produção com 5% de oxigênio dissolvido.
Title in English
Detection and cloning of 1,3-propanediol biosynthesis genes from glycerol in Klebsiella pneumoniae GLC29.
Keywords in English
1,3-propanediol
Klebsiella pneumoniae
Biodiesel
Glycerol
Metabolic flux analysis
Molecular biology
Abstract in English
The 1,3-propanediol is a compound produced from glycerol fermentation by bacteria such as Klebsiella and which can be used for various applications in industry, such as the production of PTT (polytrimethylene terephthalate). This work focused on the production of 1,3-propanediol using genes encoding the enzymes of the pathway taken from a Klebsiella strain previously isolated (GLC29). The genes were inserted into a plasmid and in the host strain of Escherichia coli with changes in the affinity of the enzyme isocitrate dehydrogenase for its cofactor. The enzyme was modified in the genome of the bacteria using the cofactor NAD instead of the original NADP. Preliminary tests on shaker showed an improvement of production by the mutated strains in comparison to the wild Escherichia coli. In biorreactor cultivation, varying the concentration of dissolved oxygen in the medium, the mutant strain showed a production 25% higher than the wild type strain in the condition with 5% of dissolved oxygen.
 
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Release Date
2020-12-04
Publishing Date
2018-12-05
 
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