A novel benzodiazepine metabolite of an isolate of Penicillium aurantiogriseum from Yugoslavia has been characterized from spectral and biosynthetic evidence. The compound, anacine [l}, has been produced on the 60-liter fermenter scale afcerextensive sporulation induced by batched Ca", and its biosynthesis was preceded by that of the diketopiperazine fructigenine Am, for which additional spectral and biosynthetic data have been obtained.