Additionally, lactyl-CoA was not detected in the extracellular compartment of breast cancer cells. Molecular docking and MD simulations predicted that lactate could serve as a substrate for acetyl-CoA synthetase besides its known substrate acetate. These projections helped to hypothesize that acetyl-CoA could have the potential to convert lactate into lactyl-CoA in the tumor microenvironment and the ensuing lactylation-mediated epigenetic process. Furthermore, molecular interaction studies helped to propose that HAT p300 may serve as a potential enzyme that can use lactyl-CoA to transfer the lactate group for the lactylation of histones and other target proteins. The nature of the data is based on in vitro detection of lactate and molecular docking and MD simulations, and these findings are novel and could be a meaningful incremental step to resolve gaps in the lactylation and tumor microenvironment.