[Objective] ZntR is a metal regulatory protein, which can catalyze the transcriptional activation of zinc efflux genes, thus controlling the intracellular Zn(II) from toxic. However, the effect of zntR on bacterial biological function remains unclear.[Methods] In this study, we constructed Aeromonas hydrophila (A.h) ∆zntR deletion strain and ∆zntR::zntR rescued strain to evaluate the biofilm formation ability, hemolytic activity, motility ability, the characters of responding to metal ion stress and other physiological phenotypes.[Results] The results showed that zntR deletion strain was sensitive to zinc and chromium stress and tolerant to cobalt ion stress, meanwhile, the biofilm formation ability was decreased and the movement ability was enhanced. These phenotypes could be recovered in the rescue strains. Furthermore, the differential protein expression between wild-type strain and zntR deletion strain was compared by DIA based quantitative proteomics. Results showed that zntR may also be involved in the regulation of two-component system, bacterial chemotaxis and other metabolic pathways.[Conclusion] The above research can provide theoretical basis for further study on the regulation mechanism of ZntR transcription factors in bacterial physiological function.