Entomopathogenic fungi are unique owing to their versatile ability to produce many bioactive compounds and from the dependence of their morphological differentiation on the presence of insect-derived materials. An entomopathogenic fungus, Metarhizium anisopliae HF293, was found to show insect-material-dependent production of antibacterial compounds, which were purified to homogeneity from 10-d culture broth when the production reached maximum. Two compounds were isolated: the major compound was determined to be helvolic acid and the minor one was a novel derivative of helvolic acid (1,2-dihydrohelvolic acid). Discovery of a novel bioactive compound indicated that insect-derived material would be a useful factor for enhancing the diversity of compounds produced by entomopathogenic fungi.