Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a classic glycolytic enzyme that plays important roles in various cellular processes. Here, we report the sequence and transcriptional analyses of a regulated gene (gpdh1) encoding GAPDH in the entomopathogenic fungus Metarhizium anisopliae, a well-characterized biocontrol agent of a wide range of arthropod pests. Transcript and protein analyses of the gpdh1 showed a carbohydrate-dependent expression pattern in response to different carbon sources. A demonstration that GAPDH is localized at the cell surface is presented, and assays with insect wings show that this protein has adhesion-like activity. These results imply that GAPDH adhesion to the wing surface is specific and may play a role in the binding of conidia to a host. Our observations indicate new roles for GAPDH both physiologically and during the entomopathogen-host interaction.