Caracterización de una actividad D-malato oxidasa y estudio de los sistemas de defensa frente a DNAs exógenos en "Pseudomonas pseudoalcaligenes" CECT 5344

Resumen

The Gram negative bacterium “Pseudomonas pseudoalcaligenes” CECT 5344 has the ability to assimilate cyanide under alkaline conditions, which makes it a strong candidate for use in processes for the elimination of industrial waste contaminated with this compound. In a previous project, the genome of “P. pseudoalcaligenes” CECT 5344 has been sequenced and, once the chromosome has been closed, the main metabolic capacities of the bacteria have been identified. Within the metabolic studies that have been developed, a D-malate oxidase activity has been detected in the periplasm of the bacteria, which has been characterized in this work. Given the importance of D-malate, in the food industry, in subsequent works, an attempt we will try to build a biosensor with the data obtained from this enzyme. Lastly, the aim has been to generate a strain specially equipped for the construction of mutants, since this is the main system that will be used to study the functions of the different genes involved in the pathway of assimilation of cyanide. Restriction Modification Systems of this bacterium involved in the elimination of exogenous DNA have been studied, which makes it difficult to introduce modified DNAs that would serve for site-directed mutagenesis. In the genome of this bacterium, at least the Modification-Restriction systems and two other gene regions that contain the genes of the CRSPR systems (clustered regularly interspaced short palindromic repeats) have been identified that could also be involved in the protection against the insertion of exogenous genetic material.