The currently employed tools to redirect metabolic flux through a specific pathway are deletion and/or insertion of regulatory sequences or genes in the bacterial chromosome. These methods permanently modify the pathway perturbing the metabolic balance during the lag growth phase, what generally extends this process and also decreases cell viability. This project proposes the construction of a new model for controlling the purine metabolism in Bacillus subtilis aiming to reversibly increase the carbon flux through it. The genes of the purine metabolism in B. subtilis are controlled by five different riboswitches whose activity can be in principle modulated by a single sRNA (small non-regulatory RNA). The goal is to exemplary model the well-studied purine metabolism in this industrially relevant bacterium and to demonstrate that the sRNA technology is actually suitable for metabolic engineering of microbial production strains. The advantage of this innovative control concept is that gene expression can be turned on but also turned off as needed. The sRNA control system will be coupled to an autocatalytic induction system (lux-system of Vibrio fischeri), which will be able to self-monitoring. The new concept is broadly applicable and could be an innovative element in the portfolio of modern synthetic biology. (AU)
Articles published in Agência FAPESP Newsletter about the research grant:
ZOCCA, VITORIA FERNANDA BERTOLAZZI;
CORREA, GRACIELY GOMES;
LINS, MILCA RACHEL DA COSTA RIBEIRO;
DE JESUS, VICTOR NUNES;
TAVARES, LEONARDO FERRO;
AMORIM, LAURA ARAUJO DA SILVA;
KUNDLATSCH, GUILHERME ENGELBERTO;
PEDROLLI, DANIELLE BISCARO.
The CRISPR toolbox for the gram-positive model bacterium Bacillus subtilis.
CRITICAL REVIEWS IN BIOTECHNOLOGY,
Web of Science Citations: 0.