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Background  The integrity of cell wall structure is highly significant for the in vivo survival of mycobacteria. We hypothesized that changes in morphology may indicate changes in cell wall metabolism and identified an  aceE  gene mutant ( aceE -mut) which presented a deficient colony morphology on 7H10 agar by screening transposon mutagenesis in  Mycolicibacterium smegmatis , basonym  Mycobacterium smegmatis  ( M. smegmatis ). This study aimed to identify the functional role of  aceE  gene in cell wall biosynthesis in  M. smegmatis .    Results  We observed that the colony morphology of  aceE -mut was quite different, smaller and smoother on the solid culture medium than the wild-type (WT) strain during the transposon library screening of  M. smegmatis . Notably, in contrast with the WT, which aggregates and forms biofilm, the  aceE -mut lost its ability of growing aggregately and biofilm formation, which are two very important features of mycobacteria. The morphological changes in the  aceE -mut strain were further confirmed by electron microscopy which indicated smoother and thinner cell envelope images in contrast with the rough morphology of WT strains. Additionally, the  aceE -mut was more fragile to acidic stress and exhibited a pronounced defects in entering the macrophages as compared to the WT. The analysis of mycolic acid (MA) using LC-MS indicated deficiency of alpha-MA and epoxy-MA in  aceE -mut strain whereas complementation of the  aceE -mut with a wild-type  aceE  gene restored the composition of MA.    Conclusions  Over all, this study indicates that  aceE  gene plays a significant role in the mycolic acid synthesis and affects the colony morphology, biofilm formation of  M. smegmatis  and bacteria invasion of macrophage.

The aceE involves in mycolic acid synthesis and biofilm formation in Mycobacterium smegmatis