UV mutagenesis and enzyme inhibitors as tools to elucidate the late biosynthesis of the spirobisnaphthalenes

The metabolite pattern of UV mutants of the spirobisnaphthalene producing fungus F-24'707 by TLC and HPLC analysis has been investigated. Mutants with differences in colony morphology or colour compared to the parent strain were isolated. Cultivation in shaking flasks and P flasks showed differences in the metabolite pattern of some of the strains. Furthermore, enzyme inhibitors were used to block the spirobisnaphthalene biosynthesis of the parent strain at different steps. Feeding of precursors and intermediates of cladospirone bisepoxide (15) led to a two-fold increase of the production of 15. From these data and preceding biosynthetic studies we deduced a general pathway for the biosynthesis of all spirobisnaphthalenes of the fungus F-24'707. This enables us to present the hypothesis that all bisnaphthalenes described so far are produced using a common pathway with only a few intermediates as central branching points.