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The natural red food colorants carmine (E120) and carminic acid are currently produced from scale insects. The access to raw material is limited and current production is sensitive to fluctuation in weather conditions. A cheaper and more stable supply is therefore desirable. Here we present the first proof-of-concept of heterologous microbial production of carminic acid in  Aspergillus nidulans  by developing a semi-natural biosynthetic pathway. Formation of the tricyclic core of carminic acid is achieved via a two-step process wherein a plant type III polyketide synthase (PKS) forms a non-reduced linear octaketide, which subsequently is folded into the desired flavokermesic acid anthrone (FKA) structure by a cyclase and a aromatase from a bacterial type II PKS system. The formed FKA is oxidized to flavokermesic acid and kermesic acid, catalyzed by endogenous  A .  nidulans  monooxygenases, and further converted to dcII and carminic acid by the  Dactylopius coccus  C-glucosyltransferase DcUGT2. The establishment of a functional biosynthetic carminic acid pathway in  A .  nidulans  serves as an important step towards industrial-scale production of carminic acid via liquid-state fermentation using a microbial cell factory.

Heterologous production of the widely used natural food colorant carminic acid in Aspergillus nidulans