Temporal separation of steps in the biosynthesis of verruculogen

A mixture of 2[3H] mevalonate and U[14C] proline, given to Penicillium simplicissimum on days 2, 3, 4, 5 or 6 of stationary liquid culture, gave rise to radiolabelled verruculogen in which the ratio of 3H to 14C measured on day 8 increased from 2.0:1 to 9.5:1 over the period of administration of radiolabelled precursors. This suggests temporal separation of the formation of the diketopiperazine moiety and its subsequent prenylation. The putative precursor diketopiperazine, cyclo-L-tryptophanyl-L-proline, was isolated from 21 day culture broth.