Defects are generally considered to be effective and flexible in the catalytic reactions of lithium-sulfur batteries. However, the influence of the defect concentration on catalysis remains ambiguous. In this work, molybdenum sulfide with different sulfur vacancy concentrations is comprehensively modulated, showing that the defect level and the adsorption-catalytic performance result in a volcano relationship. Moreover, density functional theory and in situ experiments reveal that the optimal level of sulfur defects can effectively increase the binding energy between molybdenum sulfide and lithium polysulfides (LiPSs), lower the energy barrier of the LiPS conversion reaction, and promote the kinetics of LiS bidirectional catalytic reaction. The lower bidirectional catalytic performance incited by excessive or deficient sulfur defects is mainly due to the deformed geometrical structures and reduced adsorption of key LiPSs on the catalyst surface. This work underscores the imperative of controlling the defect content and provides a potential approach to the commercialization of lithium-sulfur batteries.