The effects of microbial colonization and biofilm formation on microplastics in the marine and coastal environments have aroused global concern recently. However, the simultaneous influences of exposure time and depth on biofilm formation, and subsequently on the properties variations of microplastics is less studied. In this study, polyethylene (PE) film was exposed at three depths (2 m, 6 m, and 12 m) for three time periods (30 days, 75 days, and 135 days) in the coastal seawater of Yellow Sea, China. The results show that the total amount of biofilms markedly increased with exposure time, but decreased with water depth. Typical morphologies and compositions of biofilms such as coccus-, rod-, disc-shaped bacteria and filaments, as well as a dense layer of extracellular polymeric substances were observed on the surfaces of the PE microplastics. Biofilm formation could decrease the hydrophobicity of PE microplastics, and increase the abundances of hydrophilic C−O and CO groups on the surface of PE. Alphaproteobacteria, Gammaproteobacteria and Bacteroidia were identified as the core microbiome of the PE associated biofilms, while the dominant bacteria families vary from the early to the late phases of the biofilm formation. Our results indicate that microplastics associated biofilms could affect the environmental processes and fates of microplastics in the marine and coastal environment. Display omitted •Biofilm formation and its influences on PE properties were investigated.•The thickness of biofilms on PE increases with exposure time but decreases with depth.•Biofilms could decrease the hydrophobicity and change the functional groups of PE.•The dominant PE colonizing microbial community varies during the biofilm formation.