Three metabolites, sporothrins A, B, and C (1-3), were isolated from the mangrove endophytic fungus Sporothrix sp. (#4335). Their structures were identified by the spectral data and X-ray diffractive techniques, with compound 1 showing strong inhibition of acetylcholine esterase. 1,3,6,8- Tetrahydroxynaphthalene (T4HN) was deduced as one of the precursors in the biosynthesis of 1 and 2. In a primary biosynthesis gene study, the partial gene fragment obtained with the LC1-Im/2c-Im primer pair was shown to be closely related to genes encoding T4HN synthase. The deduced protein sequences were highly homologous to the ketosynthase domains of other fungal PKS genes involved in T4HN biosynthesis. ©2009 American Chemical Society.