In silico analysis of phag-like protein in ralstonia eutropha H16, potentially involved in Polyhydroxyalkanoates synthesis
DOI:
https://doi.org/10.33571/rpolitec.v15n29a5Keywords:
Polyhydroxyalkanoates, Ralstonia eutropha H16, non-related carbon sources.Abstract
Polyhydroxyalkanoates (PHA) are synthesised by bacteria as carbon storage material. Carbon is metabolised through either glycolysis, β-oxidation or fatty acid de novo synthesis. The latter pathway allows the use of non-related carbon sources such as acetate, gluconate or glycerol -commonly present in agro-industrial residues- for PHA synthesis through the protein PhaG. The gene that codifies for this protein has not yet been found in the genome of Ralstonia eutropha H16, which is the model organism for PHA production and which can use these carbon sources to produce PHA. By comparison to already known PhaG proteins, a PhaG-like protein was found codified by gene H16_0147. In order to confirm the presence of this gene, primers were designed and a PCR standardized, giving a product with the expected base pairs length. This is the first study that shows the presence and the characteristics of a PhaG-like protein in R. eutropha H16 and represents the first step for the identification of a connection between fatty acid de novo synthesis and PHA synthesis in this model bacterium. Further studies are necessary to confirm the potential relationship of the gene product of H16_0147 gene with the PHA metabolism.
Los polihidroxialcanoatos (PHA) son sintetizados por las bacterias como material de reserva de carbono. El carbono es metabolizado a través de la ruta de la glicólisis, de la β-oxidación o de la ruta de síntesis de ácidos grasos de novo. La última ruta mencionada, permite el uso de fuentes de carbono no relacionadas como el acetato, el gluconato o el glicerol -comúnmente presentes en residuos agroindustriales-, para la síntesis de PHA a través de la proteína PhaG. El gen que codifica esta proteína no ha sido aún encontrado en el genoma de Ralstonia eutropha H16, que es el organismo modelo en la producción de PHA y que puede utilizar estas fuentes de carbono para producirlo. A través de la comparación con proteínas PhaG ya conocidas, una proteína similar a PhaG, fue encontrada siendo codificada por el gen H16_0147. Para confirmar la presencia de este gen, se diseñaron primers y se estandarizó una PCR, que dio como resultado un producto con el número de pares de base esperado. Este es el primer estudio que muestra la presencia y características de una proteína similar a PhaG en R. eutropha H16 y representa el primer paso en la identificación de una conección entre la síntesis de ácidos grasos de novo y la síntesis de PHA en esta bacteria modelo. Más estudios son necesarios para confirmar la relación potencial entre el producto del gen H16_0147 con el metabolismo de los PHA.
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