Edwardsiella tarda Raw sequence reads. Edwardsiella tarda

Metabolic state-reprogramming approach reverses antibiotic resistance via boosting depressed metabolic pathways that contribute to antibiotic resistance. Logically, the approach may be used to explore previously unknown metabolic pathways that contribute to antibiotic resistance, especially those neglected. Since depressed pyruvate cycle is a characteristic feature in antibiotic-resistant bacteria, here, pyruvate reprogramming was performed to reverse the resistance of multidrug-resistant Edwardsiella tarda. Surprisingly, a pyruvate-regulated glutathione system via boosting glycine, serine and threonine metabolism and cysteine and methionine metabolism played a key role in the reversing. This involved in pyruvate-depressed glutathione and -promoted glutathione oxidized, which was attributed to the elevated glutathione peroxidase and depressed glutathione reductase that was inhibited by glycine. This regulation inhibited ROS degradation and thereby elevated ROS to eliminate E. tarda. Loss of metB and gpx, gor of the metabolic pathways caused the elevated and decreased resistance, respectively, in vitro and in vivo, supporting a pyruvate-cysteine-glutathione system/glycine-ROS metabolic pathway. The role of this metabolic pathway in drug resistance and reprogramming reversal was demonstrated in lab-evolved gentamicin-resistant E. tarda and other clinically isolated multidrug-resistant and carbapenem-resistant pathogens. Thus, a less studied antibiotic resistance metabolic pathway is revealed and its reversal is established.