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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2013.tde-17052014-093733
Document
Author
Full name
Gabriela Cazonato Lozano
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2013
Supervisor
Committee
Gomez, José Gregorio Cabrera (President)
Galhardo, Rodrigo da Silva
Marques, Marilis do Valle
Title in Portuguese
Avaliação do papel de genes envolvidos na mobilização de poli-3-hidroxibutirato em linhagens recombinantes de Escherichia coli.
Keywords in Portuguese
Escherichia coli
Genética bacteriana
Poliester
Recombinação genética
Abstract in Portuguese
O P3HB é um tipo de poliéster sintetizado por bactérias como reserva de carbono e energia, e mobilizado na escassez destes. Um estudo com mutantes de Burkholderia sacchari, indicou o envolvimento de PhaZa1 e LonA na mobilização de P3HB. Este estudo avaliou o papel de PhaZa1 e LonA neste processo. A complementação heteróloga dos mutantes de B. sacchari, a partir dos genes phaZa1 e lonA de Ralstonia eutropha, restabeleceu a capacidade de mobilização dessas cepas, confirmando o envolvimento de seus produtos no processo. A partir de cepas recombinantes de Escherichia coli, abrigando tanto os genes de acúmulo de P3HB como de mobilização de R. eutropha, obteve-se cepas abrigando os genes phaZa1 e lonA isoladamente e outra abrigando os dois genes simultaneamente. Quando separados, tanto phaZa1 quanto lonA, não apresentaram papel significante na mobilização porém, quando os dois genes são expressos simultaneamente, as taxas de mobilização atingem mais de 50%, indicando que deva ter uma interação entre PhaZa1 e LonA para que o processo de mobilização seja efetivo.
Title in English
Evaluation of the role of genes involved in mobilization of poly-3-hydroxybutyrate in recombinant strains of Escherichia coli.
Keywords in English
Escherichia coli
Bacterial genetics
Genetic recombination
Polyester
Abstract in English
The P3HB is a type of polyester synthesized by bacteria like carbon and energy source, and is mobilizated when there is a shortage of these. A study with mutants of Burkholderia sacchari, showed the involvement of PhaZa1 and LonA in mobilization of P3HB. The present study evaluated the role of PhaZa1 and LonA in this process. The heterologous complementation of mutants of B. sacchari, with phaZa1 and lonA genes from Ralstonia eutropha, reestablished the mobilization capacity in these strains, confirming the involvement of their products in this process. From the recombinant strain of Escherichia coli, harboring accumulation and mobilization genes from R. eutropha, we obtained strains harboring phaZa1 and lonA genes singly and another harboring both genes simultaneously. When expressed singly, both phaZa1 as lonA, had no significant role in mobilization but, when both genes were expressed simultaneously, the rates of mobilization reached more than 50%, appointing that an interaction must occur between PhaZa1 and LonA for the mobilization process to be effective.
 
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Publishing Date
2014-05-21
 
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