Abstract: Objective To evaluate the effect of SMU.2055 gene on acid resistance of Streptococcus mutans. Methods A SMU.2055- dificient mutant strain of S. mutans was constructed using homologous recombination technique. The growth of the wild-type and mutant strains was monitored in both normal and acidic conditions. The lethal pH level, glycolysis, proton permeability, cell permeability and biofilm formation of the two strains were compared. Results PCR and sequence analyses verified the successful construction of the SMU.2055-dificient mutant strain. The growth and biofilm formation capacity of the mutant strain were obviously lowered in both normal and acidic conditions. The mutant strain also showed increased lethal pH level, proton permeability, and cell permeability with impaired H+-ATPase activity in acidic conditions, but its minimum glycolytic pH remained unaffected. Conclusion The SMU.2055-deficient S. mutans mutant exhibits a lowered acid resistance, which affects the growth, lethal pH, proton permeability, H+-ATPase activity, cell permeability and biofilm formation but not the minimum glycolytic pH of the mutant strain.