The continued emergence and global spread of bacterial antibiotic resistance has fuelled the search for novel antimicrobial agents and resistance-modifying compounds. Manuka honey has both antimicrobial properties and the ability to increase the efficacy of FDA-approved antibiotic drugs. Compared to other types of honey, manuka honey contains elevated levels of methylglyoxal (MGO), a small molecule that contributes to its antibacterial activity. Manuka honey has shown particular promise for the treatment of antibiotic-resistant Gram-positive organisms such as methicillin-resistant Staphylococcus aureus. Linezolid is an oxazolidinone antibiotic used in the treatment of infections caused by a range of Gram-positive pathogens. Here, we demonstrate that manuka honey, as well as MGO in isolation, increases the sensitivity of S. aureus to linezolid in both agar diffusion and broth microdilution assays. This synergistic interaction is mediated in part by increased intracellular accumulation of linezolid in the presence of MGO.