The taxonomic position of strain JCM 2765(T) isolated from fermented cane molasses in Thailand was reinvestigated. Strain JCM 2765(T) was originally identified as representing Lactobacillus buchneri on the basis of biochemical and physiological characteristics. In the present study, 16S rRNA gene sequence analysis of strain JCM 2765(T) demonstrated a low level of similarity with the type strain of L. buchneri (92.5 %) and high levels with those of Lactobacillus collinoides (97.6 %) and Lactobacillus paracollinoides (98.0 %). Ribotyping was applied to investigate the relationships between strain JCM 2765(T), L. collinoides and L. paracollinoides. The dendrogram based on ribotyping patterns showed one cluster for six strains of L. paracollinoides, and that strain JCM 2765(T) and L. collinoides JCM 1123(T) were each independent. Based on additional phenotypic findings and DNA-DNA hybridization results, strain JCM 2765(T) is considered to represent a novel species of the genus Lactobacillus, for which the name Lactobacillus similis sp. nov. is proposed. The type strain is JCM 2765(T) (=LMG 23904(T)).