Sucrose synthase (SUS) plays a crucial role in the growth and development of plants, as well as in the response to abiotic stress and the formation of plant quality. To explore the effect of LbaSUS expression on soluble sugar accumulation in fruits, the members of the LbaSUS gene family were identified using bioinformatic tools. The physicochemical properties, gene structure, conserved motifs, phylogeny and collinearity were analyzed. The correlation between the expression of the LbaSUS genes and soluble sugar content was analyzed via quantitative real-time PCR (qRT-PCR) combined with the dynamic changes in sucrose content during fruit development. In total, six LbaSUS genes were identified based on Lycium barbarum genome, encoding proteins ranging from 739 to 918 amino acids, which were predominantly localized in the cytoplasm and unevenly distributed across five chromosomes. Phylogenetic analysis revealed that the LbaSUS gene family members were clustered into three subgroups, namely SUS I, SUS II and SUS III, and members within each subgroup shared similar gene structures and conserved motifs. Additionally, a large number of cis-acting elements associated with plant growth and development, hormone response and stress tolerance were detected in the LbaSUS family members. qRT-PCR analysis indicated that the expression levels of LbaSUS1, LbaSUS5 and LbaSUS6 gradually decreased with fruit ripening. The expression of LbaSUS2 and LbaSUS3 increased initially and then declined. The expression of LbaSUS4 showed no significant change. Subcellular localization analysis indicated that LbaSUS3 protein is localized in the chloroplast. Correlation analysis revealed a significant positive correlation between the expression level of LbaSUS3 and sucrose content. Collectively, these findings lay a foundation for further functional studies of SUS genes in Lycium barbarum.