Repressed Central Carbon Metabolism and Its Effect on Related Metabolic Pathways in Cefoperazone/Sulbactam-Resistant Pseudomonas aeruginosa

Abstract

Metabolic shift and antibiotic resistance have been reported in Pseudomonas aeruginosa. However, the global metabolic characteristics remain largely unknown. The present study characterizes the central carbon metabolism and its effect on other metabolic pathways in cefoperazone-sulbactam (SCF)-resistant P. aeruginosa (PA-RSCF). GC-MS-based metabolomics shows a repressed central carbon metabolism in PA-RSCF, which is confirmed by measuring expression of genes and activity of enzymes in the metabolism. Furthermore, expression of the genes that encode the enzymes for the first step of fatty acid biosynthesis, glutamate metabolism, and electron transport chain is reduced, confirmed by their enzymatic activity assay, and the key enzyme for riboflavin metabolism is also reduced, indicating the decreased metabolic flux to the four related metabolic pathways. Moreover, the role of the reduced riboflavin metabolism, being related to ROS generation, in SCF resistance is explored. Exogenous H2O2 potentiates SCF-mediated killing in a dose-dependent manner, suggesting that the decreased ROS resulted from the reduced riboflavin metabolism that contributed to the resistance. These results indicate that the repressed central carbon metabolism and related riboflavin metabolism contribute to SCF resistance, but increasing ROS can restore SCF sensitivity. These findings characterize the repressed central carbon metabolism and its effect on other metabolic pathways as the global metabolic features in PA-RSCF.

Riboflavin Targets the Cellular Metabolic and Ribosomal Pathways of Candida albicans In Vitro and Exhibits Efficacy against Oropharyngeal Candidiasis

“…The production of ROS may connect total metabolism via thiamine metabolism, RF metabolism, and ribosome metabolism caused by exogenous RF ( Fig. 8 ) ( 48 – 51 ). More specifically, exogenous RF targets the thiamine and RF metabolic pathways, thus disrupting central carbon metabolism.…”

“…This further aggravates the disorder in thiamine metabolism, since histidine is the substrate for thiamine synthesis. Moreover, ROS are capable of inducing protein damage and interacting with endogenous RF and thiamine metabolism ( 35 , 48 – 51 ). The accumulating pyruvate may defend against ROS ( 36 ).…”

Exogenous D-ribose promotes gentamicin treatment of several drug-resistant Salmonella

“…Central carbon metabolism, involving glycolysis, the pentose phosphate pathway, and the TCA cycle, is the core component of cell metabolism, responsible for converting nutrients into biomass to maintain life ( Baughn and Rhee, 2014 ; Barbier et al, 2018 ; Dolan et al, 2020 ). Disrupted central carbon metabolism and energy metabolism result in drug inactivation in bacterial cells ( Chen et al, 2022 ). While the TCA cycle was not affected in SCH-R, the fluctuation of amino acid metabolism and other metabolic pathways, as well as the qPCR results, suggested that the TCA cycle was disrupted.…”

“…Coincidentally，both purine and pyrimidine metabolism were seriously impacted in SCH-R and the pentose phosphate pathway and alanine, aspartate, and glutamate metabolism were shown to directly affect purine metabolism through 5-phosphoribosyl diphosphate and 5-phosphoribosylamine, respectively. Furthermore, it is worth noting that nutrients can also contribute to riboflavin metabolism through the pentose phosphate pathway, leading to ROS production, a cause of bacterial death induced by bactericidal antibiotics ( Kohanski et al, 2007 ; Chen et al, 2022 ). Whether purine and pyrimidine metabolism are involved in the inactivation of gentamicin, and whether there is a certain co-action between them and central carbon metabolism mediating the resistance of SCH-R to gentamicin remains to be further studied.…”

“…Hence, metabolomics is a potential tool to explore the acquisition of antimicrobial resistance by bacteria ( Kok et al, 2022 ). A GC–MS-based metabolomics study on cefoperazone sulbactam (SCF)-resistant P. aeruginosa (PA-R SCF ) found that the metabolic mechanism for resistance was the inhibition of central carbon metabolism ( Chen et al, 2022 ). Another metabolomics study on Escherichia coli carrying the mcr-1 gene found that mcr-1 -mediated colistin resistance was associated with a disruption in glycerophospholipid metabolism and LPS biosynthesis as well as the accumulation of the substrate PEA ( Li et al, 2019 ).…”