Solithromycin (SOL), a fourth-generation macrolide and ketolide, has been reported to have robust antibacterial activity against a wide spectrum of Gram-positive bacteria. However, the impact of SOL on planktonic growth and biofilm formation of clinical enterococcus isolates remains unclear. In this study, 276 Enterococcus faecalis isolates and 122 Enterococcus faecium were retrospectively collected from a tertiary hospital from China. SOL against clinical isolates of enterococci from China were evaluated the antimicrobial activity in comparison with erythromycin, and explore its relationship with the clonality, virulence genes and resistance mechanism of these isolates. Our data showed that the MICs of SOL against clinical E. faecalis and E. faecium isolates from China were ≤4 and ≤8 mg l⁻¹, respectively. ST16 and ST179 were regarded as the risk factor to SOL resistance in E. faecalis. SOL could inhibit but not eradicate the biofilm formation of E. faecalis. The bactericidal effects of SOL against E. faecalis and E. faecium were demonstrated to be similar to linezolid and vancomycin using time-kill assays. In conclusion, SOL showed significantly enhanced antibacterial activity against clinical isolates of E. faecalis and E. faecium from China in comparison to erythromycin. Furthermore, SOL could inhibit the biofilm formation of E. faecalis and have the similar bactericidal ability as linezolid and vancomycin against both E. faecalis and E. faecium.