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Rapid dissemination of the plasmid-born polymyxin resistance gene  mcr-1  poses a critical medical challenge. MCR-1 expression is tightly controlled and imposes a fitness cost on the bacteria. We used growth studies and metabolomics to examine growth and metabolic changes within  E. coli  TOP10 at 8 and 24 h in response to different levels of expression of  mcr-1 . Induction of  mcr-1  greatly increased expression at 8 h and markedly reduced bacterial growth; membrane disruption and cell lysis were evident at this time. At 24 h, the expression of  mcr-1  dramatically declined with restored growth and membrane integrity, indicating regulation of  mcr-1  expression in bacteria to maintain membrane homeostasis. Intermediates of peptide and lipid biosynthesis were the most commonly affected metabolites when  mcr-1  was overexpressed in  E. coli . Cell wall biosynthesis was dramatically affected with the accumulation of lipids including fatty acids, glycerophospholipids and lysophosphatidylethanolamines, especially at 8 h. In contrast, levels of intermediate metabolites of peptides, amino sugars, carbohydrates and nucleotide metabolism and secondary metabolites significantly decreased. Moreover, the over-expression of  mcr-1  resulted in a prolonged reduction in intermediates associated with pentose phosphate pathway and pantothenate and CoA biosynthesis. These findings indicate that over-expression of  mcr-1  results in global metabolic perturbations that mainly involve disruption to the bacterial membrane, pentose phosphate pathway as well as pantothenate and CoA biosynthesis.

Metabolic Perturbations Caused by the Over-Expression of mcr-1 in Escherichia coli