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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2018.tde-04122018-142151
Document
Author
Full name
Diogo Pinetti Marquezoni
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2012
Supervisor
Committee
Schenberg, Ana Clara Guerrini (President)
Rivera, Irma Nelly Gutierrez
Severino, Divinomar
Title in Portuguese
Clonagem e expressão do gene da bacteriorodopsina em Cupriavidus necator.
Keywords in Portuguese
Cupriavidus necator
Bacteriorodopsina
Expressão heteróloga
Polihidroxialcanoatos
Abstract in Portuguese
Polihidroxialcanoatos (PHAs) são polímeros produzidos por diversas bactérias como material de reserva de carbono e energia, apresentando propriedades termoplásticas comparáveis às dos plásticos de origem petroquímica, além da vantagem de ser totalmente biodegradáveis. A bactéria Cupriavidus necator DSMZ 545 é considerada o organismo modelo para a produção de PHA, possuindo a capacidade de utilizar CO2 como fonte de carbono, porém existem sérias limitações para o seu cultivo autotrófico. Visando otimizar a capacidade autotrófica desta bactéria, foi realizada neste trabalho a clonagem e expressão do gene da bacteriorodopsina (bop) de Halobacterium salinarum em C. necator. Esta proteína de membrana é capaz de converter a energia luminosa em um gradiente de prótons, que pode ser utilizado pela célula para produzir ATP. Obtiveram-se clones recombinantes de C. necator, que, sob iluminação, apresentaram maior produção de ATP, assim como produção aumentada de PHA, sem alterações no crescimento celular.
Title in English
Cloning and expression of the bacteriorhodopsin gene in Cupriavidus necator.
Keywords in English
Cupriavidus necator
Bacteriorhodopsin
Heterologous expression
Polyhydroxyalkanoates
Abstract in English
Polihydroxyalcanoates (PHAs) are polymers produced by several bacteria as carbon and energy storage materials, which show comparable thermoplastic properties with plastics from petrochemical origin and present the advantage of being completely biodegradable. The bacterium Cupriavidus necator DSM 545 is considered as a model organism for PHA production, being able to produce PHA utilizing CO2 as the carbon source, although there are severe limitations for its autotrophic cultivation. In the present work, with the purpose of optimizing the autotrophic capability of this bacterium, the cloning and expression of the Halobacterium salinarum bacteriorhodopsin gene (bop) was undertaken in C. necator. This membrane protein is able to convert light energy in a proton gradient that can be utilized by the cell for ATP production. Upon illumination, the C. necator recombinant clones showed an increase in ATP production, as well as an increase in PHA production, and no alterations in cell growth.
 
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Publishing Date
2018-12-04
 
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