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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Controle da composição do copolímero P3HB-co-3HHx por indução gradativa da expressão dos genes phaA e phaB em Pseudomonas sp. LFM461. / Composition control of P3HB-co-3HHx through gradative expression of phaA and phaB genes in Pseudomonas sp. LFM461.

Cespedes, Lucas Garbini 25 October 2016 (has links)
Os copolímeros de 3-hidroxibutirato e 3-hidroxihexanoato (P3HB-co-3HHx) são da família dos polihidroxialcanoatos (PHA), materiais termoplásticos e biodegradáveis acumulados por bactérias a partir de fontes de carbono renováveis. O P3HB-co-3HHx desperta interesse industrial para frações de 3HHx menores que 20 mol%, assemelhando-se ao polietileno de baixa densidade. Neste projeto, criou-se um sistema genético para controlar a composição do copolímero P3HB-co-3HHx pela indução de genes da biossíntese de precursores 3HB. Baseado no promotor lac foram construídas cinco versões do plasmídeo de controle de copolímero (pCC) utilizando o promotor Lac. Porém, mesmo com a versão mais aprimorada, não foi possível o controle de composição de P3HB-co-3HHx em Pseudomonas sp. LFM461. Através de experimentos de atividade enzimática e RT-qPCR do cDNA do gene phaA e phaB, foi possível indicar que o problema está na impossibilidade do promotor Lac de promover expressão dos genes de biossíntese de 3HB presentes nos pCC. / Polyhydroxyalkanoates (PHA) are polyesters material accumulated by bacteria which has thermoplastic and biodegradable proprieties and can be produced from renewable feedstocks. Copolymers of 3-hydroxybutirate and 3-hydroxyhexanoate (P3HB-co¬-3HHx) which contain less than 20mol% of 3HHx have being researched for its industrial proprieties. Recently, our laboratory has been researching a Pseudomonas sp. strain, LFM461, which can produce high 3HHx content when hosting PHA biosynthesis genes from Aeromonas. In this project, it was design a genetic system for control of P3HB-co-3HHX composition through induction of 3HB monomers biosynthesis. Thus, five versions of a copolymer control plasmid (pCC) was built based on Lac promoter. However, even through improvements on stability and expression profile of pCC it was not possible to stablish an assay of successful PHA composition control in Pseudomonas sp. LFM461. By enzymatic activity and RT-qPCR of cDNA experiments we have indications of problems of Lac promoter on driving the expression of 3HB genes in pCC.

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