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Lycopene cyclization to gamma-carotene and then to beta-carotene is partially blocked in Phycomyces by the presence of car R mutations in heterokaryosis or by the addition of 2-(4-chlorophenyl)thiotriethylamine . HCl to the medium. We have quantitatively determined the carotenes synthesized by Phycomyces under conditions of partially blocked cyclization and in the presence of either car A mutations in heterokaryosis or polyoxyethylenesorbitan monooleate (Tween-80) in the medium. We conclude that transfer of intermediate substrates between the enzyme aggregates involved in carotenogenesis does occur in the wild type but not in heterokaryons for car A mutations, and is facilitated by Tween-80.

Phycomyces strain C5, carrying mutation carB10, accumulates phytoene instead of beta-carotene. Heterokaryons containing C5 nuclei and different other nuclei carrying the wild type carB allele accumulate significant amounts of phytofluene, zota-carotene and neurosporene. From quantitative analyses of carotenes and nuclear proportions in the heterokaryons we conclude that four copies of the carB gene product, assembled in an enzyme complex, act sequentially in the conversion of phytoene to lycopene.