Streptococcus suis is one of the most important bacterial pathogens of pigs, it is also an important zoonotic pathogen. It can withstand environmental stress and respond rapidly to stress. However, the regulatory mechanisms involved in this process are not fully elucidated. MnmE is a multi-domain GTPase that is highly conserved in bacteria and eukaryotes, driving tRNA modification during translation. In this study, we constructed an mnmE deletion strain (ΔmnmE) and a complementary strain (CΔmnmE) using the wild-type strain 05ZYH33 of S. suis, and systematically studied the characteristics and functions of MnmE in S. suis through phenotypic and multi-omics analyses. Phenotypic analysis showed that the mnmE strain exhibited growth defects, reduced virulence, decreased environmental tolerance, reduced capacity for metal ion transport, and changes in protein synthesis capabilities. Multi-omics results indicated that cellular processes related to carbohydrate metabolism, transcription and translation, and inorganic ion transport and metabolism were affected, which corroborated the phenotypic analysis.