Moonlighting proteins, characterized by their multiple autonomous functions, have been detected in bacteria. Surprisingly, many of these proteins are conserved and involved in metabolic pathway or the cell stress response. They localise to the bacterial surface to take on additional activities, which have been hypothesised to contribute to bacterial virulence or bacterial benefit. In this review, we compare the functions of moonlighting proteins in bacteria, describe the structural basis of moonlighting functions, and summarise the regulation of secretion and localisation of moonlighting proteins.