Biotechnological exploitation of Pseudomonas putida Lego lizing and refactoring...
Biotechnological exploitation of Pseudomonas putida Lego lizing and refactoring central metabolic blocks through rational genome engineering
"Pseudomonads include a diverse set of bacteria with metabolic versatility and genetic plasticity that enable their survival in a broad range of environments. Many members of this family are able to both degrade toxic compounds an...
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Descripción del proyecto
"Pseudomonads include a diverse set of bacteria with metabolic versatility and genetic plasticity that enable their survival in a broad range of environments. Many members of this family are able to both degrade toxic compounds and efficiently produce high value bioproducts, and they are of interest for bioremediation endeavors as well as in bulk production of biochemicals. The present project aims at undertaking a deep genetic and metabolic engineering of Pseudomonas putida KT2440 in order to develop this strain as a biological chassis of reference for genetic and metabolic (re-)programming of bacterial catalysts à la carte. This comprises not only a fundamental (Systems Biology) inspection of P. putida, but also its streamlining into an useful catalytic vehicle for an extensive range of biotechnological applications through rational, systemic refactoring of its versatile metabolic pathways and its stress-resistance abilities. In particular, the objectives of this work include: [1] rational construction of P. putida strains in which the chromosome has been edited and deleted of most unnecessary genomic elements, [2] introduction and optimization of an efficient glycolytic catabolic pathway in thereby obtained strains, and [3] strengthening the stress resistance of these P. putida chasses by genetic outsourcing of oxidative stress-tolerance functions from Deinococcus radiodurans. Using state-of-the art methodologies, these heavily refactored bacterial constructs will be subject to a wide variety of physiological, metabolic, and genetic analysis under various physiological regimes in order to ascertain and quantify the interplay between physicochemical stress and catalytic efficiency. By undertaking these tasks, robust scaffolds for designing biocatalysts with enhanced performance under operating conditions will be obtained, with relevant information about some of their basic metabolic and genetic properties as a sound knowledge base."