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Microplastics (MPs) can adsorb toxic chemicals in biological or environmental matrixes and thus influence their behavior and availability. In order to investigate how the combined pollution of MPs and toxic organic chemical influence microbial growth and metabolism, Escherichia coli (E. coli) was grown in a complex, well-defined media and treated with polystyrene microplastics (PS MPs) and dichloro-diphenyl-tricgloroethane (DDT) at human relevant concentration levels. In vivo metabolites captured by a novel solid phase microextraction (SPME) probe, were used to reflect the metabolic dysregulation of E. coli under different pollution stresses. Results showed that the toxic effect of DDT displayed a distinct dose-dependent phenomenon while the existence of PS decreased the growth and metabolic interference effect of DDT on E. coli. Adsorption results revealed a mechanism that PS weakened the adverse impact of DDT by decreasing its free concentration in the treated culture media. Tricarboxylic acid (TCA) cycle related enzymes activities and antioxidant defense related substances of E. coli also proved the mechanism. The current study is believed to broaden our understanding of the ecotoxicity of MPs with toxic organic chemicals on microorganism.