Gram-negative bacteria possess a double-membrane envelope that serves as a robust barrier against environmental stress and antibiotics. While the transport of phospholipids synthesized at the inner membrane (IM) to the outer membrane (OM) is critical for maintaining OM integrity and functionality, the molecular mechanisms governing this process remain incompletely understood. Members of the AsmA-like protein family in E. coli have been implicated in lipid transfer to the OM, with the three largest proteins—TamB, YhdP, and YdbH—proposed as redundant lipid transporters. Of these, TamB associates with the outer membrane protein TamA to form the Translocation and Assembly Module (TAM), a complex also implicated in outer membrane protein assembly. Here, we use a combination of structural modelling, coarse grained and all-atom molecular dynamics simulations to investigate the lipid transfer ability of TAM and other members of the AsmA-like family. We show that lipids can spontaneously enter and traverse the large periplasmic spanning groove of TamB from the IM. Lipid dissociation at the OM is impeded when TamA is in complex with TamB but occurs spontaneously in structures of TamB alone. Finally, we also demonstrate that all six members of the AsmA family in E. coli can accommodate lipids within their hydrophobic grooves.