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Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2015.tde-11062015-103546
Document
Author
Full name
Thandara Garcia Ravelli
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2015
Supervisor
Committee
Silva, Luiziana Ferreira da (President)
Acosta, Maria Beatriz Riveron
Carvalho, Joao Carlos Monteiro de
Title in Portuguese
Seleção de genes codificadores de PHA sintases para a construção de recombinantes em Burkholderia sacchari e Pseudomonas sp e avaliação da produção de polihidroxialcanoatos com diferentes composições monoméricas.
Keywords in Portuguese
Burkholderia
Pseudomonas
Metagenômica
PHA
PHA sintase
Abstract in Portuguese
Polihidroxialcanoatos (PHA) são poliésteres acumulados por diversas bactérias a partir de fontes renováveis e são termoplásticos, biodegradáveis e biocompatíveis. A variabilidade da composição monomérica de PHA determina suas propriedades mecânicas e permite seu uso em diversas aplicações. A PHA sintase é a enzima responsável pela polimerização do PHA. O objetivo deste trabalho foi à busca por genes codificadores desta enzima, construção e avaliação de recombinantes portando tais genes. Inicialmente buscaram-se novos genes de PHA sintase a partir de clones de uma biblioteca metagenômica previamente detectados por PCR como positivos para algum tipo de PHA sintase. Posteriormente buscou-se PHA sintases de classe III e construiram-se recombinantes de Pseudomonas sp e B. sacchari pela introdução de genes de C. vinosum (phaECCv). Nas duas recombinantes, os genes inseridos foram capazes de aumentar a fração de 3HHx em relação a linhagem selvagem, quando se utilizou glicose e hexanoato como fontes de carbono.
Title in English
Selection of PHA synthases coding genes for the construction of recombinant Burkholderia sacchari and Pseudômonas sp and evaluation of polyhydroxyalkanoate production with different monomer compositions.
Keywords in English
Burkholderia
Pseudomonas
Metagenomics
PHA
PHA synthase
Abstract in English
Polyhydroxyalkanoates (PHA) are polyesters accumulated from renewable sources by several bacteria and are thermoplastic, biodegradable and biocompatible. The variability of the PHA monomer composition determines its mechanical properties and allows their use in many applications. PHA synthase is the enzyme responsible for the polymerization of PHA. The objective was to search for genes encoding this enzyme, construction and the assessment of recombinant bacteria carrying such genes. Initially a screening of PHA synthase genes was made from a metagenomic library clones previously identified as positive by PCR for any type of PHA synthase. Later a search for class III PHA synthases was made as a construction of a recombinant Pseudomonas sp and B. sacchari by introducing genes of C. vinosum (phaECCv). In the two recombinant strains, the genes inserted were able to increase the fraction of 3HHx compared to the wild strain when glucose and hexanoate was used as carbon sources.
 
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Publishing Date
2015-06-11
 
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