Derarrollo de herramientas de biología sintética para la bacteria no modelo Burkholderia sacchari: un nueva plataforma para una producción más limpia

Abstract

The metabolic diversity in microorganisms can provide the basis to create new biochemical products. However, most metabolic engineering projects use a handful of established model organisms and, therefore, a challenge to take advantage of the potential of novel microbial functions is the ability to express new genes or directly use non-model organisms. Genetic manipulation of non-model microorganisms is still a challenge due to the specific nuances of the organism that make universal molecular genetic tools difficult. However, in recent years, unprecedented advances have been made in synthetic biology and the development of molecular genetic tools. That is why the objective of this study is to develop useful tools of synthetic biology to enhance the use of the non-model bacterium Burkholderia sacchari, as an alternative to biological production, since it has been proven that it can synthesize several high-value products using different sources of carbon. In this work we used the BglBrix plasmids, a standard that includes all the basic biological parts necessary for a vector to be efficient, stable and fulfill its function within the cell. The transformation of Burkholderia sacchari cells with the BglBrix plasmids was carried out by chemical method using RbCl, obtaining high transformation efficiency. Dose-induction assays were performed to evaluate the expression of fluorescent proteins at a maximum concentration of inducer.