Surface-exposed virulence factors of Gram-positive bacteria are anchored to the cell wall peptidoglycan by sortase enzymes, with the length of the cell wall cross-bridge influencing envelope architecture. Ovococcal bacteria typically have mono or dipeptide cross-bridges, while coccoid-shaped Staphylococcus aureus has five glycine residues. In ovococcoid Enterococcus faecalis, the cross-bridge consists of two L-Alanines. To explore the impact of cross-bridge length and cross-linking on E. faecalis cell envelope biogenesis, we deleted FemB, the enzyme responsible for adding the terminal L-Alanine, leading to shorter cross-bridges and fewer cross-linkages. FemB mutants exhibited altered growth and morphology, appearing larger and more spherical. Unlike the parental strain, which divides in parallel, FemB mutants showed irregular septation, dysregulated peptidoglycan organization, and asymmetry between daughter cells. To understand these defects, we generated femB suppressor mutants that overcame the growth issues. Whole genome sequencing of these mutants identified mutations in a hypothetical gene and genes involved in cell wall stress responses and synthesis, suggesting their role in compensating for division defects caused by the shortened cross-bridge. Transcriptomic and phenotypic analyses indicate that oxidative stress through reactive oxygen species (ROS) production occurs in the FemB mutant, potentially explaining the altered cell division, morphology, and growth. This study demonstrates that FemB is crucial for maintaining canonical cell division along parallel planes, and its disruption triggers a ROS-associated stress response in E. faecalis.